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28 September 2021, Volume 41 Issue 9 Previous Issue   
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Innovative research team achievement column
Neonatal surgery topics
Basic research
Clinical research
Review
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Innovative research team achievement column

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A novel antithrombotic antibody targeting the binding sites of the coagulation factor FⅨa-FⅧa complex Tian-yao SUN, Shi-feng JIANG, Qin XU, Jun-ling LIU, Su-ying DANG, Xue-mei FAN 2021, 41 (9):  1133-1141.  doi: 10.3969/j.issn.1674-8115.2021.09.001 Abstract ( 755 )   PDF (6502KB) ( 1021 )   ·To prepare a monoclonal antibody targeting coagulation factor Ⅸa (FⅨa), a key factor in endogenous coagulation pathways, and study its antithrombotic roles and mechanisms. ·Immunization of mice, hybridoma technology, cell expression and purification were used to prepare the anti-FⅨa monoclonal antibodies of high purity. The monoclonal antibodies with high affinity for FⅨa were screened by enzyme-linked immunosorbent assay. The activated partial thromboplastin time (APTT) and prothrombin time (PT) were used to evaluate the antithrombotic effects of the monoclonal antibody. And then chromogenic substrate method was used to detect the effect of monoclonal antibody on the enzyme activity of FⅨa. The method of computer simulation of protein-protein docking was adopted to predict the possible binding sites between FⅨa and the antibody, and the binding site, was verified by competitive experiments (indirectly through the chromatic substrate method). ·FⅨa-4, an anti-FⅨa monoclonal antibody with high affinity, was generated. Although FⅨa-4 did not affect PT and the enzyme activity of FⅨa, it significantly prolonged APTT to 88.8 s, which was 3.5 times of the control group (25.5 s), in a concentration-gradient dependence. The protein-protein docking prediction results revealed that FⅨa-4 did not directly bind to substrate catalytic sites of FⅨa, but occupied the binding region of FⅨa and FⅧa. Competitive experiments further verified the above results. FⅨa-4 inhibited FⅩa production in a dose-dependent manner, almost completely inhibiting FⅩa production at the concentrations of 400 pmol/L, and FⅧa could correct the inhibition effect of the antibody up to nearly 50%. ·The monoclonal antibody against FⅨa-4 is obtained. FⅨa-4 competes with FⅧa to bind FⅨa, and inhibits the conversion of FⅩ to FⅩa which is catalyzed by FⅧa-FⅨa complex; it plays an antithrombotic role mainly by inhibiting endogenous coagulation pathway. Figures and Tables | References | Related Articles | Metrics

Neonatal surgery topics

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Factors related to prognosis of portoenterostomy for type Ⅲ biliary atresia Geng-feng JI, Zhi-bo ZHANG 2021, 41 (9):  1142-1146.  doi: 10.3969/j.issn.1674-8115.2021.09.002 Abstract ( 476 )   PDF (1150KB) ( 633 )   ·To explore the factors related to the jaundice clearance and native liver survival after portoenterostomy (Kasai's operation) in the children with type Ⅲ biliary atresia. ·The clinical data of the patients diagnosed as having type Ⅲ biliary atresia, who underwent Kasai's operation in Shengjing Hospital of China Medical University from January 2012 to January 2018, were analyzed retrospectively. According to the prognosis, the children were divided into jaundice clearance group (JC group, which was divided further into early and late JC group) and jaundice non-clearing group (JOC group), or native liver survival group and non-native liver survival group (liver transplantation/death). T/U test, χ2 test, Kaplan-Meier survival analysis, and multivariate Cox regression analysis were employed to analyze the prognostic factors. ·A total of 96 patients were enrolled in this study, including 48 males and 48 females. The average age of operation was (62.1±18.9) d, and the average follow-up time was (16.2±20.3) months. Direct bilirubin decreased to normal within 6 months postoperatively in 61 cases (63.5%). Compared with the JOC group, the indirect bilirubin (IBIL) level was higher and the albumin level was lower in the JC group before operation (both P<0.05). The result of Kaplan-Meier survival curve analysis showed that the native liver survival rates at 3, 6, 12 and 24 months after operation were 92.3%, 70.9%, 57.8% and 55.8%, respectively. The native liver survival rate of the JC group was higher than that of the JOC group (P=0.000). The rate was also higher in the early JC group when compared with the late JC group (P=0.001). Multivariate Cox regression analysis revealed that gender of male [P=0.049, RR=2.163 (95%CI 1.076?4.797)] and jaundice clearance [P=0.001, RR=11.488 (95%CI 2.726?48.415)] were the protective factors of 2-year native liver survival rate. ·Preoperative IBIL and albumin levels may affect jaundice clearance in the children with type Ⅲ biliary atresia after portoenterostomy; gender of male and jaundice clearance are protective factors for 2-year native liver survival in these children. Figures and Tables | References | Related Articles | Metrics

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Study on the correlation between the expression of miR-146a-5p and the severity of intestinal injury in neonatal necrotizing enterocolitis Jiang-long CHEN, Tong CHEN, Zhi-bao LÜ, Xue-li WANG, Qing-feng SHENG 2021, 41 (9):  1147-1153.  doi: 10.3969/j.issn.1674-8115.2021.09.003 Abstract ( 564 )   PDF (5829KB) ( 625 )   ·To detect the expression level of miR-146a-5p in the intestinal tract of neonatal necrotizing enterocolitis (NEC), and analyze its correlation with the severity of intestinal injury in children with NEC. ·The clinical data of children with NEC and intestinal atresia (IA) admitted to the Department of General Surgery of Shanghai Children's Hospital, Shanghai Jiao Tong University from January 2014 to December 2018 were collected, respectively. According to the location of intestinal tissue samples, children with NEC were divided into NEC-inflamed group and NEC-unaffected group. The intestinal tissue samples of children with IA were used as control group (i.e. IA group). Newborn C57BL/6J mice aged 5-7 days were randomly divided into NEC model group and control group. The former group was treated by modeling, while the latter was not. After the mice were killed, the terminal ileum tissue samples were taken. Hematoxylin-eosin staining was performed on the tissue samples of newborn mice and children in each group, and histopathological grade of mice was performed. The expression level of miR-146a-5p in the intestine of the newborn mice and the children in each group were detected by in situ hybridization, and the differences of expression between the two groups were compared, respectively. The correlation between the expression level of miR-146a-5p and histopathological grade in intestinal tract of children with NEC was analyzed. The distribution of children with miR-146a-5p high expression (positive cells≥100) and low expression (positive cells <100) in different clinical data was compared. ·① Compared with the control group, the pathological grade and the expression level of miR-146a-5p (P=0.000) in the terminal ileum of NEC model mice were higher. ② Compared with IA group and NEC-unaffected group, the expression level of miR-146a-5p was higher in NEC-inflamed group (both P=0.000). And the expression level of miR-146a-5p was positively correlated with the pathological grade of intestinal tissues in children with NEC (P=0.015, r=0.578). ③ Compared with miR-146a-5p low expression, the number of children with intestinal resection length ≥5 cm in miR-146a-5p high expression was more (P=0.005). ·The high expression of miR-146a-5p is related to the severity of intestinal injury in NEC, suggesting that miR-146a-5p is involved in the occurrence and development of NEC. Figures and Tables | References | Related Articles | Metrics

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Expression characteristics of silent information regulator transcript 1 in intestinal tissues of neonatal necrotizing enterocolitis Rui CHEN, Yun ZHAO, Xiao-xia ZHAO, Dong MA, Yi-jiang HAN, Deng-ming LAI, Wei-zhong GU, Jin-fa TOU 2021, 41 (9):  1154-1161.  doi: 10.3969/j.issn.1674-8115.2021.09.004 Abstract ( 642 )   PDF (4204KB) ( 661 )   ·To explore the expression characteristics of silent information regulator transcript 1 (SIRT1) in intestinal tissues of necrotizing enterocolitis (NEC), and preliminarily discuss the effect and possible mechanisms of SIRT1 in NEC. ·From June 2018 to October 2020, 80 children with NEC who were treated by neonatal surgery in the Children's Hospital, Zhejiang University School of Medicine were divided into observation group and control group. The tissue samples of the observation group were inflammatory necrotic intestinal tubes, while those of the control group were incised intestinal tubes. The NEC children in the control group were with intestinal strictures after conservative treatment, and then were treated by surgery again. The clinical data of the two groups were collected 24 h before surgery, including procalcitonin (PCT), hypersensitive C reactive protein (hs-CRP), cytokines [interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ)] in the serum, gender, gestational age and so on. The expression characteristics of SIRT1, nuclear factor-κB (NF-κB), transforming growth factor β1 (TGF-β1) and Smad3 proteins of the tissues in the two groups were detected by immunohistochemistry. IEC-6 cells of small intestinal epithelium of SD female rats were cultured in vitro. After SIRT1 expression in IEC-6 cells was inhibited by small interfering RNA (siRNA) technology, the protein expression of NF-κB was detected by Western blotting and the effects of SIRT1 on IEC-6 cells were detected by Cell Counting Kit-8 (CCK8) and Transwell migration assay. ·There was no significant difference in gender, gestational age, birth weight and delivery mode between the two groups. Compared with the control group, the levels of hs-CRP, PCT, IL-6 and IL-10 in the serum in the observation group increased within 24 h before surgery (all P=0.000).Compared with the margin tissues of narrow intestines in the control group, the positive expression of SIRT1 in NEC necrotic intestinal tissues in the observation group showed low expression, and the positive expression of NF-κB was overexpressed (both P=0.000). There was no significant difference in the expression of Smad3 and TGF-β1 between the two groups. The expression of SIRT1 in NEC necrotic intestinal tissue was negatively correlated with the expression of NF-κB (r=-0.592, P=0.000). After inhibiting the mRNA and protein expression of SIRT1, the relative expression of NF-κB in IEC-6 cells was increased, and the proliferation and migration ability of the cells was significantly decreased. ·The mechanism for the SIRT1 reducing progression of NEC may be that SIRT1 can inhibit the expression of NF-κB to reduce the expression of inflammatory factors in NEC, and another possible mechanism may be that SIRT1 can protect intestinal epithelial cells by promoting cell proliferation and migration. Figures and Tables | References | Related Articles | Metrics

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Laparoscopic diagnosis and treatment of high jejunal atresia and stenosis in neonates Bao-hong ZHAO, Hong-xia REN, Xiao-xia WU, Yuan-yuan JIN, Wen-yue LIU, Liang ZHAO, Hui ZHANG, Xin GUO, Xue SUN 2021, 41 (9):  1162-1168.  doi: 10.3969/j.issn.1674-8115.2021.09.005 Abstract ( 554 )   PDF (2950KB) ( 893 )   ·To investigate the effect of laparoscope on the treatment of high jejunal atresia and stenosis in neonates. ·From January 2009 to December 2019, 90 neonates with high jejunal atresia and stenosis were operated on Children′s Hospital of Shanxi province. According to whether laparoscopic surgery was used or not, they were divided into laparoscopic group (n=66) and laparotomy group (n=24). In the laparoscopic group, 27 cases of type Ⅰ atresia and stenosis less than 10 cm from Treitz ligament were anastomosed under laparoscopy, and 39 cases of jejunal atresia and stenosis 10?30 cm from the Treitz ligament, underwent extraperitoneal anastomosis by expanded trocar hole with the aid of single-site laparoscope. Traditional laparotomy exploratory incision was used in the laparotomy group. The operation time, the first postoperative oral feeding time, postoperative full oral feeding time [postoperative oral feeding volume reaching 150 mL/(kg·d)], the length of hospital stay, and mortality and incidence of complications were compared between the two groups. ·There was no significant difference between the laparoscopic group and laparotomy group in age, gender, weight, premature infant, low birth weight, other systems malformation and pathological type. There was no significant difference in operation time between the laparoscopic group and laparotomy group [(77.0±24.0) min vs (82.0±24.0) min]. The first postoperative oral feeding time, postoperative full oral feeding time [postoperative oral feeding volume reaching 150 mL/(kg·d)], and the length of hospital stay in the laparoscopic group were shorter than those in the laparotomy group [(13.3±7.2) d, (19.1±8.1) d, (22.1±8.5) d vs (17.2±7.8) d,(23.7±10.6) d, (27.0±11.8) d, all P <0.05]. There was no significant difference between the laparoscopic group and laparotomy group in mortality (1/66 vs 2/24) and incidence of complications (7/66 vs 6/24). ·Laparoscopic surgery is a safe and effective method in the diagnosis and treatment of neonatal high jejunal atresia/stenosis. Figures and Tables | References | Related Articles | Metrics

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Evaluation of postoperative pulmonary function in children with congenital diaphragmatic hernia and analysis of influencing factors Xue-yao WANG, Wei-hua PAN, Wei-peng WANG, Quan-hua LIU, Wei XIE, Lei WANG, Jun WANG 2021, 41 (9):  1169-1174.  doi: 10.3969/j.issn.1674-8115.2021.09.006 Abstract ( 518 )   PDF (902KB) ( 379 )   ·To analyze the postoperative pulmonary function of children with congenital diaphragmatic hernia (CDH) and explore the related factors affecting the recovery of pulmonary function. ·The medical data of children with CDH who were treated in the Department of Pediatric Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, and underwent infant pulmonary function test (IPFT) from November 2016 to November 2020 were retrospectively reviewed. According to the type of ventilatory dysfunction in the examination, the children were divided into restrictive group, obstructive group, mixed group and healthy group. The differences in prenatal diagnosis, diagnostic gestational week, observed/expected lung-to-head ratio (O/E LHR), birth weight, 1 min Apgar score, hernia side, liver-up, use of patch during operation, duration of mechanical ventilation, length of hospital stay, month age at the time of IPFT and postoperative follow-up time in each group were analyzed. The pulmonary function results of the first-time test were analyzed in the diagnosis of whether the gestational age was less than 25 weeks, whether there was a liver-up and whether the patch was used during the operation. The difference between the results of the first and last test was analyzed. ·Fourty-two children were included in the analysis. A total of 96 tests were conducted in patients. Sixteen cases underwent IPFT once, and 26 cases had 2 or more tests. There were significant differences in month age at the time of IPFT and postoperative follow-up time among the restrictive (n=6), obstructive (n=68), mixed (n=9) and healthy (n=13) groups (P=0.004, P=0.002). Overall, the tidal volume per kg of body weight (TV/kg), and tidal expiratory flow 50% remaining tidal volume (TEF50%) in children with diagnosis of gestational age less than 25 weeks were lower (P=0.022, P=0.043), TEF75% and TEF50% in children with liver-up were lower (both P=0.040), and TEF50% and TEF25% in children with use of patch during operation were lower (P=0.034, P=0.020). The higher the O/E LHR, the higher the ratio of time to peak tidal expiratory flow to total expiratory time (TPEF/TE) and the ratio of volume at peak tidal expiratory flow to expiratory tidal volume (VPEF/VE) (r=0.393, P=0.005; r=0.324, P=0.023). The less the duration of mechanical ventilation, the higher the TPEF/TE and VPEF/VE (r=-0.377, P=0.000; r=-0.314, P=0.002). Within 1 month after operation, obstructive ventilatory dysfunction occurred 6 times (46.2%), and obstructive ventilatory dysfunction was found 49 times (77.8%) 3 months after operation. In patients who received IPFT twice or more, with the increase of follow-up time, TV/kg, TEF75%, TEF50% and TEF25% in the last IPFT were higher than those in the first test (P=0.001, P=0.003, P=0.001, P=0.000), and the respiratory rate decreased (P=0.010). ·The tidal respiratory function of CDH children diagnosed before the first 25 weeks of pregnancy, with liver-up and intraoperative patch use is even worse. The main manifestation of early and late postoperative ventilatory dysfunction is obstruction. With the passage of time of postoperative follow-up, the pulmonary function of children with CDH is improved. Figures and Tables | References | Related Articles | Metrics

Basic research

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Role of Tmprss6 gene in radiation-induced intestinal injury of mice Jiu-ang MAO, Zhen WENG, Xiao-yin NIU, Yang HE, Zhen-xin WANG 2021, 41 (9):  1175-1182.  doi: 10.3969/j.issn.1674-8115.2021.09.007 Abstract ( 711 )   PDF (4110KB) ( 496 )   ·To explore the effect of Tmprss6 gene on radiation-induced intestinal injury in mice. ·7?8 weeks old wild-type (WT) male mice and Tmprss6 knockout (Tmprss6-/-) mice were randomly divided into non-irradiated (IR) group and groups of 6 h, 24 h, 3.5 d and 7 d post-IR. The IR group received total abdominal X-ray irradiation (TAI) at a dose of 11.5 Gy to establish radiation intestine injury models. H-E staining was performed in the 3.5 d post-IR group, and TUNEL immunofluorescence was performed in the 6 h and 24 h post-IR groups to observe the pathological damage and apoptosis of different intestinal segments, respectively.The levels of iron of different intestinal segments were measured 0, 3.5 and 7 d after TAI, as well as superoxide dismutase (SOD), glutathione (GSH), total antioxidant capacity (T-AOC) and malondialdehyde (MDA). The correlation analysis between iron and each antioxidant index was performed. ·The length of duodenal villi and the number of crypts decreased significantly in both Tmprss6-/- mice and WT mice 3.5 d after TAI. Meanwhile, the duodenal villi length and crypt count of Tmprss6-/- mice were lower than those of WT mice (both P<0.05). The number of TUNEL-positive apoptotic cells per crypt in the duodenal segment of Tmprss6-/- mice was significantly increased compared with that of WT mice 6 and 24 h after TAI (both P<0.05). On the contrary, the length of ileum villi and the count of crypts increased significantly, while the number of TUNEL-positive apoptotic cells decreased significantly in Tmprss6-/- mice compared with that of WT mice (all P<0.05). The iron content in the duodenum of WT and Tmprss6-/-mice 7 d after TAI was significantly increased compared with that of non-irradiated mice (both P<0.05). And the iron content of the duodenum in the Tmprss6-/- mice before and after TAI was significantly higher than that of WT mice at the same time points (all P<0.05). The iron content of the ileum in WT and Tmprss6-/- mice also increased after TAI. And the increase was statistically significant at 3.5 d post-IR in the WT mice, so was at 7 d post-IR in the Tmprss6-/- mice (both P<0.05). In addition, the SOD, GSH and T-AOC levels in the duodenum of Tmprss6-/- mice were lower than those in the WT mice after TAI, while the content of MDA was higher. Among them, the difference of SOD, GSH, MDA and T-AOC levels were statistically significant 3.5 d after TAI (all P<0.05). Contrary to the duodenum, the contents of SOD, GSH, and T-AOC in the ileum of Tmprss6-/- mice were higher than those in the WT mice, while the content of MDA was lower. Among them, the content of T-AOC, GSH and MDA had a statistically significant difference 3.5 d after TAI (all P<0.05), and the content of SOD had a statistically significant difference 7 d after TAI (P=0.031). The results of correlation analysis showed that there was no significant correlation between the MDA, GSH, SOD and T-AOC levels and the iron content of the small intestine in the non-IR group. The levels of SOD and MDA in the duodenum (r=0.639, r=-0.710, both P<0.05), and T-AOC, MDA and GSH in the ileum (r=-0.817, r=-0.658, r=-0.726, all P<0.05) of WT mice were significantly correlated with their respective iron content after TAI. The levels of MDA, SOD and T-AOC in the duodenum of Tmprss6-/-mice were significantly correlated with the iron content after TAI (r=-0.606, r=0.771, r=-0.712, all P<0.05), and all the antioxidant indexes of the ileum were not significantly correlated with the iron content. ·The iron level in the small intestine is significantly correlated with the oxidative stress response after irradiation. Tmprss6 gene-deficient mice show increased duodenal iron accumulation and irradiation-induced oxidative stress, and Tmprss6 gene deletion exacerbates duodenal radiation damage. Figures and Tables | References | Related Articles | Metrics

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Whole gene expression profile analysis of miRNAs in human umbilical vein endothelial cells regulated by vascular endothelial growth factor A Li LIU, Zi-long GENG, Jia-huan CHEN, Sha-sha ZHANG, Bing ZHANG 2021, 41 (9):  1183-1189.  doi: 10.3969/j.issn.1674-8115.2021.09.008 Abstract ( 722 )   PDF (1846KB) ( 509 )   ·To analyze the changes of microRNA (miRNA) in human umbilical vein endothelial cells (HUVECs) under the stimulation of vascular endothelial growth factor A (VEGFA), in order to further explore the roles of miRNA in regulating the expression of downstream genes of VEGFA, and to find new regulatory miRNAs for angiogenesis. ·The miRNA expression profile was analyzed by using NanoString nCounter in HUVECs 0, 1, 4, and 12 h after VEGFA stimulation. The differentially expressed miRNAs with the same trend were clustered and their target genes were predicted. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze and predict the functions of these target genes, and the interaction networks were established. ·The expression of 39 miRNAs changed significantly after VEGFA stimulation in HUVECs. Bioinformatic analysis showed that there were 129 target mRNAs corresponding to the differentially expressed miRNAs which were involved in multiple biological processes and signal pathways related to angiogenesis. ·miRNAs are differentially expressed in HUVECs after stimulation by VEGFA, and miRNAs play important roles in regulating angiogenesis. MiR-107 and miR-21 may severe as candidate functional regulators of angiogenesis. Figures and Tables | References | Related Articles | Metrics

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Effects of heat shock protein 70 on cytoactive of sciatic nerve cryopreservation and nerve regeneration after allograft Yi-feng SHI, Ying-ru HUANG, Yun-xiao LIU, Song ZHANG, Hua XIAN 2021, 41 (9):  1190-1196.  doi: 10.3969/j.issn.1674-8115.2021.09.009 Abstract ( 638 )   PDF (3352KB) ( 403 )   ·To investigate the effect of heat shock protein 70 (HSP70) on cryopreserved rat sciatic nerve cell activity and nerve regeneration after allotransplantation. ·Bilateral sciatic nerves of SD rats were placed in DMEM medium and pretreated with heat stress in vitro at 37 ℃, 42 ℃ and 45 ℃ for 1 h (37 ℃ group, 42 ℃ group and 45 ℃ group, respectively). A fresh nerve group was set up as the control (Con) group. The number of nerves in each group was 36. The expression of HSP70 in sciatic nerve was detected by Western blotting. The previously described nerves were stored in a cryopreservation solution with liquid nitrogen for 4 weeks. The expressions of major histocompatibility complex class Ⅰ (MHC-Ⅰ), MHC-Ⅱ, caspase-3 and caspase-9 were determined by Western blotting. The survival of sciatic nerve cells was detected by flow cytometry. The cryopreserved sciatic nerves were cultured at 37 ℃ with 5% CO2 for 7 d. The expressions of brain-derived neurotrophic factors (BDNF) and glial cell line-derived neurotrophic factors (GDNF) were detected by Western blotting. The cryopreserved sciatic nerves and the fresh sciatic nerves of SD rats were employed to repair the 10-mm-defect of sciatic nerve in the corresponding Wistar rats in the 37 ℃-Allo group, 42 ℃-Allo group, 45 ℃-Allo group, Con-Allo group and Fresh-Allo group. Meanwhile, an isograft group was also established (Fresh-Iso group). Each group had six Wistar rats. After 20 weeks of transplantation, the compound muscle action potential (CMAP) and nerve conduction velocity (NCV) were detected using electrophysiology. Toluidine blue staining was performed to observe the number of regenerated myelinated nerve fibers. The ultrastructure of the regenerated nerves was visualized through a transmission electron microscope. ·HSP70 expression level was higher in the 42 °C group than that in the 37 °C group, 45 °C group and Con group, respectively (all P<0.05). After 4 weeks of cryopreservation, the expressions of MHC-Ⅰ and MHC-Ⅱ in the 42 ℃ group were not statistically different from those in the 37 ℃ group, 45 ℃ group and Con group, respectively. The expression levels of caspase-3 and caspase-9 in the 42 °C group were lower than those in the 37 °C group , 45 °C group and Con group, respectively, and the survival rate of sciatic nerve cells was increased (all P<0.05). After 7 d of culture, the expressions of BDNF and GDNF in the 42 ℃ group were higher than those in the 37 ℃ group, 45 ℃ group and Con group (all P=0.000), respectively. After 20 weeks of the transplantation, CMAP, NCV, the number of myelinated nerve fibers and myelin sheath thickness in the 42 °C-Allo group were superior to those in the Cont-Allo group and Fresh-Allo group (all P=0.000). ·Highly expressed HSP70 can minimize cryopreservation injury in sciatic nerves of rats, improve nerve cell viability after preservation, and promote nerve regeneration after allotransplantation. Figures and Tables | References | Related Articles | Metrics

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Construction of a metastasis prediction model of microsatellite instability-high colorectal cancer based on differentially expressed gene assembly Ying XU, Yi-min CHU, Da-ming YANG, Ji LI, Hai-qin ZHANG, Hai-xia PENG 2021, 41 (9):  1197-1206.  doi: 10.3969/j.issn.1674-8115.2021.09.010 Abstract ( 788 )   PDF (5369KB) ( 578 )   ·To explore the potential key genes and the gene expression characteristics of microsatellite instability-high (MSI-H) colorectal cancer (CRC) with metastasis at the transcriptome level, and establish a metastasis prediction gene model. ·The transcriptome data of MSI-H CRC patients was obtained from The Cancer Genome Atlas database. The patients were divided into metastatic group (21 patients) and non-metastatic group (42 patients). The differentially expressed genes (DEGs) between the two groups were analyzed by Gene Ontology (GO) and Gene Set Enrichment Analysis (GSEA) to annotate, and cluster DEGs and enrich the signaling pathways. STRING and Cytoscape were used to select the hub genes. Nomogram was drawn based on the selected DEGs. The cross validation of the model was performed by Bootstrap method. Survival analysis was done to explore the influences of each gene in the nomogram on progression-free survival (PFS) of MSI-H CRC. ·A total of 245 DEGs were obtained from the metastatic group and non-metastatic group, among which 204 genes were up-regulated and 41 genes were down-regulated. GO analysis showed that DEGs were mainly clustered in ion transmembrane transport, chloride transmembrane transport and chloride channel activity in terms of biological process and molecular function. In terms of cellular component, DEGs were mainly clustered in extracellular region and extracellular space. GSEA showed that the neuroactive ligand-receptor interaction and metabolic pathways were enriched in the up-regulated genes. The top 10 hub genes in the protein-protein interaction network of the up-regulated genes were screened by Cytoscape. The metastasis prediction gene model, which was set up based on the top 10 DEGs with the lowest adjusted P value and high physiological relevance to tumor, had certain predictive efficiency [area under curve (AUC)=0.975 for training, AUC=0.920 for validation]. The expression levels of AC078993.1 and IGLJ2 (immunoglobulin lambda joining 2) were significantly negatively correlated with PFS of MSI-H CRC (P=0.011, P=0.005). ·The changes in ion channels and extracellular environment may have important impacts on metastasis of MSI-H CRC. Neuroactive ligand-receptor interaction and metabolic pathways may be two important signaling pathways for metastasis of MSI-H CRC. A metastasis prediction gene model is established, which can provide reference for the follow-up related clinical researches. Figures and Tables | References | Related Articles | Metrics

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Expression and clinical significance of DNA polymerase θ in endometrial cancer Bei-bei XUAN, Sai-nan GONG, Jia-li LIU, Quan QUAN, Yu MENG, Xiao-ling MU 2021, 41 (9):  1207-1214.  doi: 10.3969/j.issn.1674-8115.2021.09.011 Abstract ( 689 )   PDF (3442KB) ( 355 )   ·To explore and verify the differential expression of DNA polymerase θ (POLQ) in endometrial cancer (EC) and normal endometrial tissues, and analyze its relationship with clinical characteristics of EC patients and its value in prognostic judgement. ·Gene Expression Profiling Interactive Analysis (GEPIA) website was used to retrieve the expression of POLQ in cancer tissues and normal tissues in various parts of the human body. The POLQ gene expression profile and the clinical data were downloaded from The Cancer Genome Atlas (TCGA) database. The gene expression matrix was obtained by Strawberry Perl software. The differences in the POLQ expression between EC and paracancerous tissues and the relationship between the POLQ expression and the clinical characteristics were analyzed by R software. Gene Set Enrichment Analysis (GSEA) was used to reveal the signaling pathways in which POLQ participated in EC. The clinical information of 127 patients diagnosed as having EC by histopathology, who underwent surgery in the First Affiliated Hospital of Chongqing Medical University from March 2011 to December 2014, were collected to analyze the differences in the POLQ expression among patients with different clinical characteristics. Immunohistochemical staining was performed on paraffin sections of tumor tissues from these patients to determine the expression of POLQ in the tumor tissues, and normal endometrium tissues from 22 patients with uterine fibroids were used as controls. Kaplan?Meier method and multivariate Cox regression model were used to analyze the correlation between POLQ expression and prognosis. ·It was found that POLQ expression was relatively lower in testicular germ cell tumor, but increased in most cancers through GEPIA website. The expression of POLQ in EC tissues was significantly higher than that in paracancerous tissues in the TCGA database (P=0.000), and POLQ was more expressed in the patients with the age of 60 years and above and in the highly graded patients (both P<0.05). Overall survival of the patients with high expression of POLQ was shorter (P=0.000). The results of GSEA showed that POLQ gene was mainly enriched in the base resection repair pathway, homologous recombination pathway, P53 pathway, etc. The proportions of POLQ scores ≥1 and ≥5 in the normal endometrial paraffin sections were significantly lower than those in the EC tissues (both P<0.05). The proportion of POLQ score ≥5 showed statistically significant difference among the patients with different tumor stages or different conditions of lymph node metastasis (both P=0.038). The disease-free survival and overall survival of the patients with high POLQ expression were significantly shorter than those of the patients with low POLQ expression (both P<0.05). Multivariate Cox analysis showed that POLQ score could be used as an independent prognostic indicator. ·Compared with normal endometrium, the expression of POLQ significantly increase in EC tissues, especially in patients with advanced stage and lymph node metastasis, and high expression of POLQ is an independent predictor of poor prognosis in EC patients. Figures and Tables | References | Related Articles | Metrics

Clinical research

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Association study of variation of glucagon-like peptide-1 receptor gene rs3765467 and type 2 diabetes mellitus Jing-jing ZHANG, Chao-yu ZHU, Yuan-yuan XIAO, Fu-song JIANG, Qing-ge GAO, Yun-yun FANG, Li WEI 2021, 41 (9):  1215-1221.  doi: 10.3969/j.issn.1674-8115.2021.09.012 Abstract ( 625 )   PDF (968KB) ( 395 )   ·To investigate the association of single nucleotide polymorphisms (SNPs) in the glucagon-like peptide-1 receptor (GLP-1R) gene with type 2 diabetes mellitus (T2DM). ·A total of 318 Han patients with T2DM and 145 non-diabetic controls in Shanghai were enrolled.The polymorphisms of GLP-1R gene were detected with multiplex PCR-High throughput resequencing. Differences in frequencies of genotypes and alleles, as well as other clinical variables between the two groups were analyzed. ·Twenty three SNPs were detected. The polymorphism of GLP-1R rs3765467 (G/A) was significantly associated with T2DM (P=0.005). Compared with GG genotype carriers, GA+AA genotype carriers had a lower risk of T2DM (OR=0.502, 95% CI 0.305-0.829, P=0.007), while GA+AA genotype carriers had a significantly lower risk of T2DM (OR=0.403, 95% CI 0.186-0.871, P=0.021) in the male population. Association analysis of clinical feature showed that the glycosylated albumin level of GA+AA genotype carriers was lower than that of GG genotype carriers (P=0.048). ·GLP-1R rs3765467 polymorphism is associated with T2DM in Shanghai Han population. Minor allele A contributes to maintaining blood glucose homeostasis and is a protective factor for T2DM, especially in men. Figures and Tables | References | Related Articles | Metrics

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Clinical characteristics and survival analysis of 52 cases with intracranial atypical teratoid/rhabdoid tumor Yan-li XU, Wen-bin GUAN, Wei-wei MAO, Chuan-ying ZHU, Qin ZHANG, Xiao-jun YUAN, Zhen TAN 2021, 41 (9):  1222-1227.  doi: 10.3969/j.issn.1674-8115.2021.09.013 Abstract ( 863 )   PDF (1809KB) ( 785 )   ·To summarize the clinical characteristics and survival status of children with intracranial atypical teratoid/rhabdoid tumor (AT/RT) diagnosed and treated in recent 8 years, and analyze the survival-related factors, so as to provide basis for further therapeutic stratification. ·From October 2012 to January 2020, 52 AT/RT children treated in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine were selected. Their clinical data and follow-up were recorded and their 3-year progression-free survival (PFS) and 3-year overall survival (OS) were analyzed by Kaplan-Meier method. According to the age of the children, they were divided into ≥3-year-old group and <3-year-old group. The clinical characteristics and survival status of the two groups were compared. Univariate and multivariate Cox proportional hazards regression models were used to analyze the impact of clinical characteristics on overall survival. The significance of immunohistochemical indicators in children with different survival outcomes was explored. ·There were 34 males and 18 females in AT/RT children, 65.4% of which were younger than 3 years old, and the average age of onset was (35.2±28.2) months. The median survival time was 7.7 months, and 3-year PFS and 3-year OS was (29.3±7.6)% and (30.4±7.6)%, respectively. There were significant differences between the two groups in receiving radiotherapy and chemotherapy, serious complications, 3-year PFS and 3-year OS (all P<0.05). Univariate Cox regression analysis showed that the survival risk of children, who was less than 3 years old and underwent primary tumor invading the spinal canal, tumor metastasis, and partial resection, without radiotherapy or chemotherapy, was increasing (all P<0.05). Multivariate Cox regression analysis showed that tumor metastasis was an independent risk factor for overall survival of children (P=0.024). The results from immunohistochemistry showed that the positive expression of SYN in survival children was significantly higher than that in dead children (P=0.003). ·Children younger than 3 years old with AT/RT have more severe complications and lower overall survival rate, so it is necessary to implement more accurate therapeutic stratification and diversified treatment methods. Figures and Tables | References | Related Articles | Metrics

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Establishment and evaluation of screening model of minimal hepatic encephalopathy in patients with liver cirrhosis Zhu-ping QIAN, Yan YANG 2021, 41 (9):  1228-1232.  doi: 10.3969/j.issn.1674-8115.2021.09.014 Abstract ( 547 )   PDF (2667KB) ( 389 )   ·To establish and evaluate a screening model of liver cirrhosis patients complicated with minimal hepatic encephalopathy (MHE). ·A total of 404 patients with liver cirrhosis who were hospitalized from June 2017 to November 2019 were selected as the research subjects, and the clinical data of them were collected. Based on Logistic regression analysis and artificial neural network (ANN), the MHE screening models were established, and the discriminant ability of the two models was evaluated and compared. ·The Logistic regression model showed that age, history of diabetes mellitus, infection, renal insufficiency, nutritional risk, total bilirubin>24 μmol/L, blood ammonia>47 μmol/L and international normalized ratio (INR)≥1.5 were the significant risk factors (all P<0.05). The area under the curve (AUC) of receiver operator characteristic curve (ROC curve) of ANN model and Logistic regression model were 0.814 and 0.737 (Z=4.208, P=0.000), respectively. The sensitivities were 72.4% and 69.9%, and the specificities were 76.7% and 67.8%, respectively. ·The ANN model is more effective than the Logistic regression model in MHE screening. Figures and Tables | References | Related Articles | Metrics

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Value of CT radiomic features in preoperative prediction of cervical lymph node metastasis in patients with papillary thyroid carcinoma Jun-lin HE, Qing LU, Xin XU, Shu-dong HU 2021, 41 (9):  1233-1239.  doi: 10.3969/j.issn.1674-8115.2021.09.015 Abstract ( 718 )   PDF (2188KB) ( 520 )   ·To explore the value of radiomic features of pre-contrast phase,arterial phase and venous phase in predicting preoperatively cervical lymph node metastasis (CLNM) in patients with papillary thyroid carcinoma (PTC) in multi-phase images of contrast-enhanced CT scan. ·CT images of 197 PTC patients who underwent thyroid surgery in Tinglin Hospital of Jinshan District of Shanghai from January 2017 to June 2020 were collected. 512 frames that meet the research requirement, consisting of 193 pre-contrast phases,131 arterial phases and 188 venous phases, were selected from the 3 phases of CT images 197 patients, and the CT images showing the largest length to diameter of PTC lesion were chosen for the radiomic study in each one of the 512 frames. The optimal parameters of RandomForestClassifier were selected with all the 512 frames of CT images and random forest (RF) classification model for the prediction of CLNM was established based on CT images with all the 3 phases of 124 patients who had concurrent CT images of pre-contrast phase, arterial phase and venous phase. The predictive performance of the models was estimated by area under the curve (AUC) of receiver operator characteristic curve (ROC curve) analysis. ·The RF classification models showed that the maximal average AUC of ROCs of pre-contrast phase, arterial phase and venous phase were 0.843, 0.775 and 0.783, and the corresponding predictive accuracy were 0.767, 0.695 and 0.726, respectively. Compared with the arterial phase and venous phase, the radiomic features extracted from pre-contrast phase of CT images show better performance to predict CLNM (both P=0.000). ·Radiomic features extracted from pre-contrast phase, arterial phase and venous phase of CT images can all feasibly be used to predict CLNM in patients with PTC, and radiomic features from pre-contrast phase of CT images show better performance. Figures and Tables | References | Related Articles | Metrics

Review

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Research progress of imaging markers for identifying and predicting the progression of age-related macular degeneration Yi-yang SHU, Si-qi ZHANG, Hai-yun LIU 2021, 41 (9):  1240-1245.  doi: 10.3969/j.issn.1674-8115.2021.09.016 Abstract ( 563 )   PDF (825KB) ( 752 )   With the increase in the number of age-related macular degeneration (AMD) patients, imaging markers with indicative value have been screened and applied to the initial screening of AMD, disease warning, recurrence reminder and treatment guidance to contribute to the early diagnosis and accurate treatment for AMD patients. In clinical practice, colour fundus photography, optical coherence tomography (OCT), optical coherence tomography angiography (OCTA) and fundus angiography are mostly used for image recognition and analysis. This article reviews the imaging markers for identifying and predicting the progression of early-, middle- and late-stage AMD, as well as predicting the recurrence and prognosis of AMD, aiming to provide a theoretical basis and treatment guidance for clinical judgment. References | Related Articles | Metrics

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Advances in cell wall structure and Candida albicans-host interaction Yi-kun MEI, Jing-cong TAN, An-jun WANG, Hui WANG, Ning-ning LIU 2021, 41 (9):  1246-1251.  doi: 10.3969/j.issn.1674-8115.2021.09.017 Abstract ( 1174 )   PDF (948KB) ( 1584 )   Candida albicans is an opportunistic human fungal pathogen. The mortality rate of blood infection caused by Candida albicans is as high as 40% while that of the cancer patients can even reach to 70%, which greatly increases the burden of human health. The cell wall of Candida albicans is the frontline of defense against external stress and the first contact point between fungi and host. Therefore, the cell wall is very important for fungi-host interaction and immune recognition, and is the most attractive target for antifungal therapy. Once the balance of synthesis and remodeling of cell wall of Candida albicans is broken, the molecular integrity of cell wall will be disturbed, which will finally lead to cell lysis and death. This paper reviews the characteristics of cell wall components of Candida albicans and their functions on fungi-host interaction and immune recognition mechanism, in order to provide novel research clues and theoretical basis for identification of specific antifungal targets and development of more economic and effective antifungal drugs. References | Related Articles | Metrics

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Application of Quality-of-Life Assessment Scales and psychological assessment tools to people with dry eye disease Yu-ying CAI, Yan LIU 2021, 41 (9):  1252-1255.  doi: 10.3969/j.issn.1674-8115.2021.09.018 Abstract ( 574 )   PDF (798KB) ( 1609 )   Dry eye disease (DED) is a common disease with multiple etiologies. It can affect the daily activities of patients to reduce their quality of life and interfere with their mental state. At present, for DED patients, the clinical application of Quality-of-Life Assessment Scales are more, but the summary research of professional psychological assessment tools is less. On the basis of extensive review at home and abroad, this paper summarizes and explores the application of Quality-of-Life Assessment Scales and psychological assessment tools to DED patients, in order to provide suggestions and guidance for the clinical diagnosis and treatment and nursing work of DED. References | Related Articles | Metrics

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Research progress in the relationship between endosomal sorting complexes required for transport and autophagy Yi LI, Da-wei SUN, De-rong CUI 2021, 41 (9):  1256-1260.  doi: 10.3969/j.issn.1674-8115.2021.09.019 Abstract ( 715 )   PDF (1111KB) ( 825 )   Autophagy is an intracellular degradation pathway, which can transport a large number of cytoplasmic, damaged organelles and other substrates to lysosomes for degradation to maintain intracellular homeostasis. In the process of autophagy, the integrity of autophagy flow should be ensured. Many factors affect the smooth autophagy flow, including the relationship between endosomal sorting complexes required for transport (ESCRT) and autophagy. ESCRT is mainly responsible for membrane shearing, mediating cell membrane remodeling and division, and plays an important role in the process of endocytosis and cytokinesis. In recent years, many studies have shown that ESCRT mutants generally present an accumulation of autophagosomes, suggesting that autophagy flow is affected. In-depth studies indicate that ESCRT is involved in the regulation of several processes of different types of autophagy, and its subunits also interact with autophagy-related proteins. This article reviews the related research in recent years. Figures and Tables | References | Related Articles | Metrics

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Progress in the influence of prenatal exposure to phthalates on placental function and its mechanism Qian-long ZHANG, Ying TIAN 2021, 41 (9):  1261-1266.  doi: 10.3969/j.issn.1674-8115.2021.09.020 Abstract ( 615 )   PDF (915KB) ( 711 )   Phthalates (PAEs) are widespread in life and are a class of environmental endocrine disruptors (EEDs). The abundance of hormone receptors makes the placenta highly sensitive to EEDs. During pregnancy, women can be exposed to PAEs, and their metabolites can pass through the placenta and affect its function by interfering with hormone receptors.The dysfunction of placenta will result in fetal growth restriction or, if more severe, fetal death. Functional placental disruptions linked to phthalates exposures include invasion/migration, oxidative stress, cell differentiation/apoptosis, hormone secretion and lipid accumulation.In this paper, the exposure level of PAEs during pregnancy is summarized, the mechanism for the effect of phthalates on placenta is reviewed, and the possible limitations of these studies are discussed, aiming to provide insights for further studies on the potential molecular mechanism through which PAEs disrupt placental function. Figures and Tables | References | Related Articles | Metrics

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Research progress of artificial oocyte activation technology in clinical application Pei-qin CHEN, Di SUN, Yue FU, Qi-meng XIAO, Ming-zhu BAI, Zhen-bo ZHANG 2021, 41 (9):  1267-1271.  doi: 10.3969/j.issn.1674-8115.2021.09.021 Abstract ( 884 )   PDF (916KB) ( 989 )   Oocyte activation deficiency (OAD) is a common cause of failure of intracytoplasmic sperm microinjection (ICSI). The occurrence of OAD is usually related to the inability of sperm to produce sufficient amount of phospholipase C-ζ (PLCζ), causing calcium (Ca2 +) oscillation to activate oocytes. In order to solve OAD and improve the success rate of ICSI, the research and clinical application of artificial oocyte activation (AOA) are becoming more and more extensive. However, its biological safety has not been fully determined. It is necessary to clarify the role of Ca2 + in oocyte activation, summarize the principles of different activators and their current status in clinical application, and explore the safety and application prospect of AOA technology. References | Related Articles | Metrics

Brief original article

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Clinical application and effect of phage on the treatment of pulmonary infection by pan-drug resistant Klebsiella pneumoniae Li-sha LI, Jian-hui LI, Bin HE, Nan-nan WU, Tong-yu ZHU, Xiao-kui GUO, Zheng-hong CHEN 2021, 41 (9):  1272-1276.  doi: 10.3969/j.issn.1674-8115.2021.09.022 Abstract ( 1022 )   PDF (1937KB) ( 504 )   ·To explore the therapeutic frequency and effect of phage treatment on pan-drug resistant Klebsiella pneumoniae by aerosolized phage. ·The patient's sputum was taken 1 h before the start of treatment and every hour during the treatment of pulmonary infection using nebulized inhaled phage. The sputum samples were subjected to bacterial isolation, purification and mass spectrometry identification. The lytic spectrum of different bacterial isolations was compared. And blood samples for bacteria were taken 1 h before the start of the treatment and 2 h and 4 h after the first treatment for bacterial culture and phage volume determination. ·The phage continued to proliferate in vivo for 2 h, and decreased after 3 h. When the second phage treatment was performed after 4 h, the pattern of phage proliferation in vivo was the same as the first time. During the third treatment, the phage was proliferated slightly in vivo. After phage treatment, Klebsiella pneumoniae was still isolated in the patient's sputum. However, the phage cleavage spectrum of the isolations was different, and the patient's clinical symptoms improved. ·Phages can effectively target infectious bacteria and kill them. The clinical symptoms of patient improve. Twice aerosolized phage treatment for the patient with pulmonary infection is enough to achieve the best effect. The isolated bacterial strain is different post phage treatment. Figures and Tables | References | Related Articles | Metrics