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    Expert forum
    Molecular diagnosis and clinical significance of 21-hydroxylase deficiency
    LÜ Yongfen, LI Pin
    2022, 42 (5):  557-561. 
    doi: 10.3969/j.issn.1674-8115.2022.05.001

    Abstract ( 757 )   HTML ( 544 )   PDF (1350KB) ( 914 )  

    21-hydroxylase deficiency is the most common type of congenital adrenal hyperplasia. Currently, diagnosis is mainly based on clinical symptoms and biochemical tests, but misdiagnosis or missed diagnosis is prone to occur and genotype analysis of patients is not possible. Testing for CYP21A2 can help confirm the diagnosis of the disease, but molecular genetic diagnosis is more complex than many other single-gene disorders due to the high variability of the 21-hydroxylase genome region. There are 95% of pathogenic mutations due to recombination between functional gene and pseudogene, with 10 of the most common variants including large deletions and and gene conversions, 8 point pathogenic variants of CYP21A2 and CYP21A1P (p.P30L, I2G, p.I172N, E6 cluster, p.V281L, F306+T, p.Q318X, and p.R356 W), and p.G110fs (8 bp deletion) of exons 3. Best practice genotyping should be polymerase chain reaction (PCR)-based sequence analysis along with multiplex ligation-dependent probe amplification. Because there is a good correlation between genotype and phenotype, genotype can be used to predict the severity of clinical phenotype and guide treatment and genetic counseling in most cases.

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    Basic research
    Effects of cryogen spray combined with polymeric hydrogel on acute burn wounds
    DONG Jiaoyun, SONG Fei, LU Shuliang, LIU Yan, TIAN Ming
    2022, 42 (5):  562-569. 
    doi: 10.3969/j.issn.1674-8115.2022.05.002

    Abstract ( 469 )   HTML ( 155 )   PDF (4898KB) ( 319 )  
    Objective

    ·To develope a medical device for first aid of burns, a new formulation of cryogen spray and polymeric hydrogel, which can quickly reduce skin temperature and has no irritation, and investigate the effect of cryogen spray combined with polymeric hydrogel on quickly blocking the continuous injury of heat source to the skin tissue, and on the early repair of wound and reduction of the area and depth of burn wound.

    Methods

    ·The cryogen spray was filled into the high-pressure tank and its cooling performance was evaluated. Polymeric hydrogel dressing was prepared and the cooling time was evaluated. The irritating effect on rat' back skin tissue was evaluated after exploring on cryogen spray combined with polymeric hydrogel continuouly for 72 h. A burn rat model was created, the rats were randomly divided into two groups with 6 in each group. The burn area and inflammatory reaction of cryogen spray combined with polymeric hydrogel treated for 24 h was compared with that of the untreated group or the tap water treatment group.

    Results

    ·Compared with tap water, the cooling effect of 15% concentration cryogen spray was the best. Within 3 minutes, the cooling rate was significantly higher than that of the tap water control group (P<0.05). The cooling effect of polymeric hydrogel can be maintained for 8 h, the temperature of 70 ℃ water can be reduced to room temperature (22 ± 2) ℃ within 2 h, and the water temperature was lower than that of the control group throughout the whole process. Both materials had no irritating effect on skin tissue, and did not cause allergic reactions such as erythema and edema of skin tissue. Compared with the untreated group and the tap water group, the eschar area of the burn wound treated with cryogen spray combined with polymeric hydrogel decreased significantly (P<0.05). H-E staining showed that compared with the untreated group and the tap water group, the number of inflammatory cells in the wound of the combined application of cryogen spray and polymeric hydrogel group decreased significantly (P<0.05).

    Conclusion

    ·The combination of cryogen spray and polymeric hydrogel shows good cooling efficiency. The combined application can reduce the eschar area, and prevent the progressive deepening of the wound and the accumulation of inflammatory cells, which is conducive to the wound healing as soon as possible in the later stage. It will not cause allergic reaction of skin tissue. It may be exploited as an effective and convenient tool for the first aid management of burn patients..

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    Effects of gingipain extracts on brain neuroinflammation in mice
    ZHANG Huanyu, JIANG Yiting, ZHU Xiaochen, HE Zhiyan, ZHOU Wei, SONG Zhongchen
    2022, 42 (5):  570-577. 
    doi: 10.3969/j.issn.1674-8115.2022.05.003

    Abstract ( 441 )   HTML ( 282 )   PDF (2736KB) ( 214 )  
    Objective

    ·To observe the effects of gingipain extracts on brain inflammation in mice.

    Methods

    ·The gingipain extracts were obtained from Porphyromonas gingivalis ATCC33277 by sonication method combined with low temperature ultracentrifugation, followed by ultrafiltration and concentration. Gingipain extracts were identified by Western blotting after protein quantification. SDS polyacrylamide gel electrophoresis was performed and then transferred the proteins to the polyvinylidene fluoride (PVDF) membrane. After blockage, the proteins were probed with lysine-specific gingipain (Kgp) antibody and arginine-gingipain (Rgp) antibody, respectively, to detect the immunogenicity of gingipain in the extracts. The acute infection model was established by intraperitoneal gingipain extracts in C57BL/6N mice. The specific fluorescent substrates Z-His-Glu-Lys-MCA for Kgp and Boc-Phe-Ser-Arg-MCA for Rgp were used to detect the activities of two gingipains in plasma at different time points after intraperitoneal injection of gingipain extracts in mice. C57BL/6N mice were randomly divided into 4 groups, which were recorded as the control group, the gingipain group, the gingipain+Kgp inhibitor (COR388) group and the COR388 group, respectively. Mice in the gingipain+COR388 group were pre-treated with COR388 for 1 h, and then intraperitoneally injected with gingipain extracts. Twenty-four hours later, the samples were collected under anesthesia and the brains were processed sequentially by procedures such as fixing, dehydrating, embedding and dissecting. The microglia and astrocytes in the mouse cortex were labeled with ionized calcium binding adapter molecule 1 antibody (anti-Iba1) and glial fibrillary acidic protein antibody (anti-GFAP), respectively, and the activation of microglia and astrocytes in the cortex of the above four groups of mice were observed through immunohistochemistry. The expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were evaluated by enzyme linked immunosorbent assay (ELISA).

    Results

    ·There were relatively clear bands around 50 000 and 70 000 in the Western blotting analysis of the gingipain extracts, suggesting that the extracts contained gingipain immunogenicity. Meanwhile, the gingipain extracts were biologically active, and the activity of gingipain in the plasma of mice showed a trend of first increasing and then decreasing after intraperitoneal injection of the gingipain extracts. The activity of plasma gingipain in mice reached a peak 4?8 h after intraperitoneal injection of gingipain extracts, and then gradually decreased. Immunohistochemistry results showed that the microglia and astrocytes in the cortex had increased volume of cell bodies and irregular protrusions, suggesting that intraperitoneal injection of gingipain extracts could activate microglia and astrocytes in the mouse brain. ELISA results showed that the expression of inflammatory factors (TNF-α and IL-1β) in cortex increased.

    Conclusion

    ·Acute infection of gingipain extracts can induce neuroinflammation in mice.

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    Effect of altered expression of long non-coding RNA-B230352I09 on proliferation and cycle of H9C2 cardiomyocytes
    XU Feixiang, WANG Sheng, XUE Mingming, TONG Chaoyang, CHEN Yumei
    2022, 42 (5):  578-582. 
    doi: 10.3969/j.issn.1674-8115.2022.05.004

    Abstract ( 396 )   HTML ( 382 )   PDF (2108KB) ( 158 )  
    Objective

    ·To investigate the effect of altered expression of long non-coding RNA (lncRNA)-B230352I09 on proliferation and cycle of H9C2 cardiomyocytes.

    Methods

    ·The lncRNA-B230352I09 overexpression vector (pcDNA-B230352I09) and the negative control vector pcDNA-negative control group (NC) were constructed and transfected into H9C2 cardiomyocytes with the Lipofectamine 3000 (Lipo3000) transfection solution. The expression of lncRNA-B230352I09 was measured by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) to verify its transfection efficiency. The H9C2 cardiomyocytes were divided into blank control group, pcDNA-NC group, and lncRNA-B230352I09 overexpression group. Cell counting kit-8 (CCK8) was used to measure the absorbance (optical density, OD) of H9C2 cardiomyocytes at a wavelength of 450 nm and draw a growth curve; 5-ethynyl-2'-deoxyuridine (EdU) was used to label proliferating H9C2 cardiomyocytes, the numbers of which were observed by fluorescence microscopy. Cardiomyocyte cycle was assessed by flow cytometry and cycle-related genes expression was measured by RT-PCR.

    Results

    ·Compared with the pcDNA-NC group, the expression level of lncRNA-B230352I09 was significantly higher in the lncRNA-B230352I09 overexpression group (P=0.000), suggesting that the lncRNA-B230352I09-transfected H9C2 cardiomyocyte model was successfully constructed. Compared with the pcDNA-NC group, the CCK8 assay showed that lncRNA-B230352I09 overexpression significantly increased the OD of H9C2 cardiomyocytes and promoted the proliferation of H9C2 cardiomyocytes with a significant time-dependent enhancement of cell proliferation (24 h: P=0.000; 48 h: P=0.000; 72 h: P=0.001). Fluorescence microscopy revealed that the proportion of EdU-labeled positive H9C2 cardiomyocytes was significantly increased in the lncRNA-B230352I09 overexpression group compared to the pcDNA-NC group. Flow cytometric analysis showed a significant increase in the proportion of H9C2 cardiomyocytes in S phase and a decrease in G1 phase in the lncRNA-B230352I09 overexpression group compared to the pcDNA-NC group (P=0.000). RT-PCR showed that the mRNA expression levels of cyclin D1 and cyclin dependent protein kinase 1 (CDK1) in the lncRNA-B230352I09 overexpression group were significantly higher compared with the pcDNA-NC group (P=0.000).

    Conclusion

    ·lncRNA-B230352I09 can enhance myocardial proliferative capacity by regulating cyclin D1 and CDK1 to promote the entry of cardiomyocytes to into S phase.

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    Establishment and evaluation of standardized steps for building a guinea pig model of auditory brainstem implantation
    ZHOU Xiang, PAN Jinxi, ZHANG Qinjie, LI Yun, CHEN Ying, TAN Haoyue, PENG Fei, HUANG Sui, TAN Zhiping, WU Hao, JIA Huan
    2022, 42 (5):  583-590. 
    doi: 10.3969/j.issn.1674-8115.2022.05.005

    Abstract ( 461 )   HTML ( 188 )   PDF (5314KB) ( 323 )  
    Objective

    ·To establish the guinea pig model of auditory brainstem implant (ABI) by modified transbulla-labyrinthine approach and assess its effectiveness and reliability.

    Methods

    ·Twenty healthy adult male albino guinea pigs with normal hearing underwent ABI surgery via modified transbulla-labyrinthine approach. During the operation, the dorsal cochlear nucleus (DCN) of the brainstem was exposed laterally and implanted with a special ABI electrode array for guinea pigs. The evoked auditory brainstem response (eABR) was recorded from DCN and compared with the eABR waveform elicited from cochlear intraoperativly. After the electrode was implanted, the eABR waveform at DCN was recorded before euthanasia at 48 h, 7 d, 30 d and 180 d after operation. Then the brain tissue containing the electrode was harvested and the electrode neural-interface was evaluated by hematoxylin-eosin staining, immunofluorescence staining and q-PCR.

    Results

    ·The eABR was successfully recorded at DCN from all animals during surgery, and the death occurred at 36 h to 48 h after operation. The waveform morphology of eABR at DCN was similar to that at cochlea. The latency of wave Ⅲ in eABR at DCN [ (0.84 ± 0.22)ms] was shorter than that of wave Ⅳ in EABR at cochlea (n=19), and the threshold difference between them was (0.11±0.14) mA (n=19). Compared with the intraoperative eABR at DCN, the average threshold increased by (0.10±0.10) mA (n=16), and the average delay of wave Ⅲ latency was (0.15±0.21) ms (n=16). Histological morphology analysis showed that the contact surface of cochlear nucleus around the electrode showed vacuolar changes at 48 h post-operation, and increased glial components were observed at the edge of DCN at 48 h, 7 d and 180 d post-operation. Results of q-PCR showed IL-1β and MMP9 were upregulated at 48 h and 7 d post-operation compared with the unimplanted electrode group, but there was no significant difference.

    Conclusion

    ·The modeling success rate of established ABI-implanted guinea pig model is high and have outstanding repeativeness, and the local gliosis is preliminarily observed after implantation.

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    lncRNA GK-IT1 influences the carcinogenesis of non-small cell lung cancer cells through regulating aldolase A
    LIU Ziyang, WANG Xiaowen, CHEN Li
    2022, 42 (5):  591-601. 
    doi: 10.3969/j.issn.1674-8115.2022.05.006

    Abstract ( 456 )   HTML ( 138 )   PDF (6407KB) ( 202 )  
    Objective

    ·To investigate the effects of long non-coding RNA (lncRNA) GK-IT1 on the carcinogenesis of non-small-cell lung cancer (NSCLC) A549 and H358 cells by regulating aldolase A (ALDOA).

    Methods

    ·The expression of GK-IT1 in 42 pairs of NSCLC tissues and adjacent tissues, and NSCLC cell lines were tested by quantitative real-time PCR (qRT-PCR). The subcellular localization of GK-IT1 in A549 and H358 cells was tested by fluorescence in situ hybridization (FISH). RNA immunoprecipitation (RIP) was used to assess the interaction of GK-IT1 and ALDOA. A549 and H358 cells were transfected with GK-IT1 small interfering RNA sequences (Si-GK-IT1 #1 and Si-GK-IT1 #2) and negative control (Si-NC), and then the two cell lines were also cotransfected with Si-GK-IT1 #2 and ALDOA overexpression plasmid. The Si-NC, Si-GK-IT1 #1, Si-GK-IT1 #2 groups were set. CCK-8 and EdU assay were used to detect cell proliferation. Transwell invasion and scratch assay were used to observe cell invasion and migration. Western blotting was carried out to verify the expression of vimentin, E-cadherin, N-cadherin and ALDOA in each experimental group.

    Results

    ·Compared with the normal tissues, the relative expression of GK-IT1 was significantly higher in NSCLC tissues. Compared with BEAS-2B cells, the expression of GK-IT1 in H358 and A549 cells was significantly higher (P<0.05). FISH assay indicated that GK-IT1 was mainly located in the cytoplasm. RIP and bioinformatics analysis suggested that GK-IT1 might interact with ALDOA. Compared with the Si-NC group, the results of CCK-8 assay showed that the cell proliferation of the Si-GK-IT1 #1 and Si-GK-IT1 #2 group was significantly inhibited (P<0.05). The results of EdU assay showed that the ratio of EdU positive cells in the Si-GK-IT1 #1 and Si-GK-IT1 #2 group was significantly lower than that in the Si-NC group (P<0.05). Compared with the Si-NC group, the results of Transwell invasion assay indicated that the invasion of the Si-GK-IT1 #1 and Si-GK-IT1 #2 group was significantly inhibited (all P=0.000). The results of cell scratch assay showed that the healed wound ratio in the Si-GK-IT1 #1 and Si-GK-IT1 #2 group was remarkably lower than that in Si-NC group (all P=0.000). GK-IT1 knockdown significantly decreased the expression of ALDOA and vimentin, while the expression of E-cadherin was increased. Overexpression of ALDOA reversed the effects of GK-IT1 silencing on cell proliferation, invasion, migration and protein expression of NSCLC cells (all P<0.05).

    Conclusion

    ·GK-IT1 could promote the proliferation, invasion and migration of NSCLC cells via regulating ALDOA.

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    Clinical research
    Reliability and accuracy analysis of root coverage esthetic score
    SUN Wentao, XIE Yufeng, SHU Rong
    2022, 42 (5):  602-608. 
    doi: 10.3969/j.issn.1674-8115.2022.05.007

    Abstract ( 595 )   HTML ( 104 )   PDF (2769KB) ( 268 )  
    Objective

    · To evaluate the reliability and accuracy of the root coverage esthetic score (RES) in clinical applications by comparing RES and the visual analogue scale (VAS) results after root coverage procedures.

    Methods

    · Thirty-two patients who visited the Department of Periodontology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January to December 2020 with a total of 105 gingiva recessions were enrolled in this study. All gingiva recessions were treated with envelope technique combined with connective tissue graft (CTG). The patients' periodontal conditions were examined 6 months after surgery and the esthetic results were scored by 2 periodontists through RES and patients themselves through VAS. Intraclass correlation coefficient (ICC) and 95% confidence interval (95% CI) were used to represent the consistency of physician RES results. The differences of keratinized gingiva (KG) and gingival recession (GR) at baseline and 6 months after surgery were compared by using t-test. Analysis of variance was used to test the differences of each index among different VAS grades. Linear regression analysis was used to evaluate the relationship between percentage of root coverage (PRC) and VAS, RES and VAS. Multiple linear regression analysis was carried out with VAS as the outcome variable and 5 indexes of RES as independent variables. All tests were bilateral tests. The difference was statistically significant when P<0.05.

    Results

    ·In this study, KG increased by (1.19 ± 1.28) mm and GR decreased by (1.77±1.21) mm 6 months after surgery. The average PRC was 81.4%±39.9%, and the percentage of complete root coverage (PCRC) was 69.5%. There was no significant difference between the RES scores of the 2 periodontists (ICC=0.95, 95%CI 0.92?0.96). KG (P=0.008), PRC(P=0.000) and PCRC (P=0.000) varied significantly between different VAS grades. The RES and VAS scores were linearly correlated (P=0.000). Multiple linear regression analysis showed that VAS scores were linearly correlated with gingival margin (GM), soft tissue texture (STT) and muco-gingival junction (MGJ) (P=0.000, P=0.000,P=0.006).

    Conclusion

    · To a certain extent, the results of RES are consistent with the patient's esthetic judgment.

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    Analysis of genetic and clinical characteristics of PMP22-associated peripheral neuropathy
    ZHU Xiaowei, ZHAN Feixia, ZHANG Chao, LIU Shihua, ZHONG Ping, CAO Li, LUAN Xinghua
    2022, 42 (5):  609-616. 
    doi: 10.3969/j.issn.1674-8115.2022.05.008

    Abstract ( 817 )   HTML ( 183 )   PDF (3321KB) ( 416 )  
    Objective

    ·To investigate the genetic and clinical features of peripheral neuropathy patients with PMP22 gene variations and the association between genotype and clinical phenotype.

    Methods

    ·A total of 162 patients with peripheral neuropathy who were treated in Department of Neurology, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University School of Medicine from 2006 to 2022 were collected. Molecular detecting techniques including combining multiplex ligation-dependent probe amplification (MLPA), whole exon sequencing (WES) and Sanger sequencing were used to comprehensively analyze the genetic variation of 162 patients, and 26 probands with PMP22 gene variations and their 33 family members were screened out. The genetic variations included duplication, deletion and point mutations. The clinical and genetic characteristics of 26 patients with PMP22-associated peripheral neuropathy were further analyzed. The clinical characteristics included clinical demographic data, clinical manifestations, electrophysiological and pathological characteristics.

    Results

    ·There were a total of 59 (37 male and 22 female) individuals with PMP22-associated peripheral neuropathy, among whom 46 patients in 13 families had positive family histories and 13 did not. The median age at onset of neuropathy was 34.0 (14.0, 49.5) years old. The initial symptom was limb weakness or limb numbness. There were 51 Charcot-Marie-Tooth type-1A (CMT1A) patients with PMP22 duplication, 6 hereditary neuropathy with liability to pressure palsies (HNPP) patients with PMP22 deletion and 2 Dejerine-Sottas syndrome (DSS) patients with PMP22 point mutations (p.S72L and p.G100V). Sixteen patients underwent electrophysiological detections. Compared with the 4 PMP22 deletion patients, the motor nerve conduction velocity (MCV), compound muscle action potential (CMAP) and sensory nerve conduction velocities (SCV) of 12 PMP22 duplication patients were obviously decreased (all P<0.05). Conduction block was found in both kinds of mutation types. Sural nerve biopsies were performed in 12 patients, including 8 PMP22 duplication, 3 PMP22 deletion and 1 PMP22 point mutation, and pathological changes such as thinning of myelinated nerve fibers, “onion bulbs” and decreased nerve fiber density were observed in varying degrees.

    Conclusion

    ·PMP22 gene-associated neuropathies can be characterized by CMT1A, HNPP and DSS with high clinical heterogeneity. Patients with demyelinating manifestations on electrophysiological detections and sural nerve biopsy need genetic testing for PMP22 gene duplication, deletion and point mutation after ruling out acquired etiology. It is available to help optimize diagnosis and genetic counseling of the disease.

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    Relationship between histopathological features, Ki-67 expression and prognosis of conjunctival melanoma
    ZHU Tianyu, ZONG Chunyan, XU Shiqiong, GE Shengfang, FAN Xianqun, JIA Renbing
    2022, 42 (5):  617-623. 
    doi: 10.3969/j.issn.1674-8115.2022.05.009

    Abstract ( 564 )   HTML ( 188 )   PDF (2191KB) ( 144 )  
    Objective

    ·To investigate the relationship between histopathological features, Ki-67 expression and prognosis of conjunctival melanoma (CoM).

    Methods

    ·Thirty-six patients with histopathologically confirmed CoM admitted to the Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine between August 2008 and September 2020 were selected, and their clinical characteristics and histopathological features data were collected to analyze their clinicopathological characteristics, Ki-67 expression and prognostic relationships.

    Results

    ·Thirty-six patients with CoM were included in this study, including 26 males and 10 females, with a mean age of 57.81 (31?82) years. The site of onset was located in the lacrimal caruncle in 11 cases (30.6%), the fornix in 8 cases (22.2%), and the palpebral conjunctiva in 15 cases (41.7%). One patient (2.8%) had T1 stage disease, 12 (33.3%) had T2 stage disease, and 23 (63.9%) had T3 stage according to the American Joint Committee on Cancer (AJCC) 8th edition staging system. During the follow-up period, local recurrence was observed in 16 patients (44.4%), distant metastases in 17 patients (47.2%) and 16 patients (44.4%) died of disease. The Breslow thickness of CoM tumors was 4.05 (0.2?15.9) mm, with 18 cases (50.0%) >4 mm. Ulceration and regression were observed in 13 cases (36.1%) and 7 cases (19.4%), respectively. Tumor-infiltrated lymphocytes (TILs) were rated as “brisk” in 4 patients (11.1%) and “nonbrisk” in 26 patients (72.2%). Three patients (8.3%) had perineural invasion and 8 patients (22.2%) had vascular invasion. Thirty-five cases (97.2%) were in the vertical growth phase, and microscopic satellitosis was observed in only one case (2.8%). Regarding the histological type, 18 cases (50.0%) were superficial spreading, 12 (33.3%) were nodular, 4 (11.1%) were lentiginous, and 2 were unknown. Ki-67 index>20% was observed in 14 patients (38.9%). Breslow thickness >4 mm indicated high risk of local recurrence (P=0.049) and disease-related death of CoM (P=0.049). Histological ulceration and regression were risk factors for local recurrence (P=0.030) and distant metastasis (P=0.046), respectively. In addition, Ki-67 index was related with local recurrence (P=0.016). Spearman correlation analysis showed that Ki-67 index was positively correlated with local recurrence (P=0.433). CoM patients with a Ki-67 index higher than 20% had a higher risk of local recurrence [hazard ratio (HR) 3.61, 95% confidence interval (CI) 1.26?10.28, Log-rank P=0.011].

    Conclusion

    ·Histopathological features and Ki-67 expression of CoM can be used to predict patient prognosis and help guide clinical treatment.

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    Observation of analgesic effect of ultrasound-guided brachial plexus combined with intercostal brachial nerve block in the operation of distal humeral fracture
    CHEN Moxi, XU Tao, GAO Xiaoyun, WANG Wenying, CHEN Yongzhu
    2022, 42 (5):  624-628. 
    doi: 10.3969/j.issn.1674-8115.2022.05.010

    Abstract ( 468 )   HTML ( 114 )   PDF (1239KB) ( 236 )  
    Objective

    ·To investigate the analgesic effect of ultrasound-guided intercostobrachial nerve (ICBN) block on patients with distal humeral fracture who need surgical treatment through posterior approach.

    Methods

    ·One hundred and forty-nine patients with distal humeral fracture were enrolled in Shanghai Sixth People's Hospital from February 2016 to March 2021. They underwent elective surgery via posterior approach under non-general anesthesia and were selected for retrospective analysis. They were divided into two groups according to the way of nerve block: 107 cases of brachial plexus block (BP group), and 42 cases of brachial plexus combined with ICBN block (ICB group). The ICB group was divided into two subgroups according to different approaches of ICBN block: 17 cases in “under pectoralis minor” group (ICB1 group) and 25 cases in “axillary region” group (ICB2 group). The general condition of the four groups, the intraoperative sufentanil dosage, visual analogue scale (VAS) pain score, sedation rate and other analgesic demand rates were analyzed.

    Results

    ·There was no significant difference in age, sex, ASA classification, BMI, operation time and bleeding volume between the BP group and ICB group (P>0.05). There was also no significant difference in these general features between the ICB1 group and ICB2 group (P>0.05). The usage of sufentanil, VAS score, sedation rate and other analgesic drug demand rates in the ICB group were lower than those in the BP group (P=0.001, P=0.000, P=0.012, P=0.018), which was statistically significant. There was no significant difference in aspects mentioned above between the two subgroups (P>0.05).

    Conclusion

    ·Compared with brachial plexus block alone, ICBN block combined with brachial plexus block can significantly enhance the analgesic effect in the operation of distal humeral fracture through posterior approach, and the analgesic effect of the two ICBN block techniques is similar.

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    Surgical treatment of moderate ischemic mitral regurgitation complicated with reduced left ventricular ejection fraction
    YAO Haoyi, LIU Yun, QIN Kaijie, ZHOU Mi, WANG Zhe, CHEN Anqing, ZHAO Qiang, LI Haiqing
    2022, 42 (5):  629-634. 
    doi: 10.3969/j.issn.1674-8115.2022.05.011

    Abstract ( 427 )   HTML ( 214 )   PDF (1262KB) ( 193 )  
    Objective

    ·To explore different surgical strategies in patients of moderate ischemic mitral regurgitation (IMR) complicated with reduced left ventricular ejection fraction (LVEF).

    Methods

    ·From May 2013 to May 2019, 31 patients diagnosed as moderate IMR with LVEF <50% received surgical treatment in Department of Cardiovascular Surgery in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. The patients were divided into off-pump coronary artery bypass grafting (OPCABG) group and CABG+mitral valve plasty (MVP) group according to whether or not mitral valve procedure was performed. Preoperative, postoperative and follow-up echocardiography data and perioperative complications were compared to evaluate the surgical efficacy.

    Results

    ·There were 12 patients in OPCABG group and 19 patients in CABG+MVP group. 1 patient died in CABG+MVP group postoperatively, which showed no significant difference in mortality (5.3% vs 0) between the groups. The degree of mitral regurgitation was significantly reduced in CABG+MVP group (P=0.000), while intensive care unit (ICU) length of stay significantly increased [3.0 (1.0, 4.0) d vs 1.0 (1.0, 2.0) d, P=0.027]. After 48.7 (30.2, 66.5) months of follow-up, there were no significant differences in mortality, degree of mitral regurgitation, LVEF and complications between the two groups.

    Conclusion

    ·OPCABG procedure treating chronic moderate IMR with reduced LVEF could be beneficial for enhanced recovery after surgery and has satisfactory early and mid-term efficacy.

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    Evidence-based medicine
    Efficacy of preventive intervention of cryotherapy on radiation-induced oral mucositis in head and neck cancer: a meta-analysis
    CHEN Weihong, HOU Lili, YANG Ling, MAO Yan, ZHANG Jinfeng
    2022, 42 (5):  635-645. 
    doi: 10.3969/j.issn.1674-8115.2022.05.012

    Abstract ( 444 )   HTML ( 218 )   PDF (3347KB) ( 360 )  
    Objective

    ·To systematically review the efficacy of cryotherapy in head and neck cancer patients with radiation-induced oral stomatitis.

    Methods

    ·PubMed, EMbase, Cochrane Library, Web of Science, CNKI, WanFang, VIP and CBM databases were searched to collect randomized controlled trials (RCTs) of cryotherapy in head and neck cancer patients with ration-induced stomatitis from inception to January 31th, 2022. Literature screening, data extraction, and quality evaluation were performed independently by two researchers. Meta-analysis was then performed by using RevMan 5.4 and Stata 16.0 software.

    Results

    ·A total of 36 RCTs were included. The results of meta-analysis showed that compared to control group, cryotherapy group had a lower total incidence of radiation-induced stomatitis [adds ratio (OR)=0.30, 95% confidence interval (CI) 0.23?0.39, P=0.000] and a lower incidence of severe radiation-induced stomatitis (OR=0.17, 95%CI 0.14?0.21, P=0.000).

    Conclusion

    ·Cryotherapy may contribute to reducing the incidence of radiation-induced stomatitis and the incidence of severe radiation-induced stomatitis.

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    Public health
    Study on influencing factors and effect evaluation of patients with acute myocardial infarction in the cardiac rehabilitation center
    XU Li, YANG Yan, CHEN Hanfen, JIANG Meng, PU Jun
    2022, 42 (5):  646-652. 
    doi: 10.3969/j.issn.1674-8115.2022.05.013

    Abstract ( 455 )   HTML ( 125 )   PDF (1338KB) ( 358 )  
    Objective

    ·To analyze the relevant influencing factors of patients with acute myocardial infarction (AMI) in the cardiac rehabilitation center (CRC), and evaluate the rehabilitation effect, in order to provide evidence for the effective development of cardiac rehabilitation.

    Methods

    ·From May 2020 to October 2021, 454 patients with moderate and high-risk AMI who received emergency percutaneous coronary intervention in Renji Hospital, Shanghai Jiao Tong University School of Medicine were selected. Their general demographic data and disease situation were collected and analyzed. Patients were divided into two groups based on whether they chose to visit the CRC. Logistic regression model was performed to explore the relevant factors of AMI patients enrolled in CRC, and to compare the changes of cardiopulmonary function and body fat measurement of AMI patients before the first visit to CRC and after 3 months in CRC after discharge.

    Results

    ·A total of 32 (7.05%) patients (group 1) were enrolled in CRC after discharging from ward, and the other patients (group 2) did not. The analysis of general demographic data and disease situation showed that there were significant differences in age, religious belief, educational background and working state (all P<0.05). Logistic regression analysis showed that age, educational background (high school / technical secondary school, college or above), occurrence of two cardiac vascular lesions and left ventricular ejection fraction (LVEF) before discharge were the protective factors of AMI patients enrolled in CRC, while religious belief was the risk factor (all P<0.05). Compared with the indexes before the first visit to CRC, the peak ventilation oxygen per kilogram (t=7.619, P=0.000) and ventilation oxygen in the anaerobic threshold per kilogram (t=5.510, P=0.000) of AMI patients enrolled in CRC 3 months after discharge were higher, while the body mass index (t=3.132, P=0.004) and waist hip rate (t=3.891, P=0.000) were lower.

    Conclusion

    ·The enrolling percentage of moderate and high-risk AMI patients in CRC is still at a low level, and age, religious belief, educational background, cardiac vascular lesions and LVEF before discharge are the influencing factors. Patients enrolling in CRC 3 months later have significant improvement in cardiopulmonary function and body fat.

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    Techniques and methods
    Development of endoscopic image classification models of Barrett's esophagus based on deep convolutional neural networks
    LIN Jiaxi, WANG Shengjia, ZHAO Xin, GAO Xin, YIN Minyue, ZHU Jinzhou
    2022, 42 (5):  653-659. 
    doi: 10.3969/j.issn.1674-8115.2022.05.014

    Abstract ( 390 )   HTML ( 141 )   PDF (2395KB) ( 220 )  
    Objective

    ·To develop classification models for endoscopic images of Barrett's esophagus using deep convolutional neural network, and evaluate its classification performance.

    Methods

    ·A total of 806 esophageal images were collected from Gastrointestinal Endoscopy Center of the First Affiliated Hospital of Soochow University and the HyperKvasir database, including 412 normal esophageal images and 394 Barrett's esophagus images. All the images were randomly divided into training set (85%) and validation set (15%). Four deep convolutional neural networks [Xception, NASNet Large (NASNetL), ResNet50V2 (ResNet) and BigTransfer (BiT)], which had been pre-trained in the ImageNet database, were used to perform transfer learning in the training set and develop endoscopic image classification models of Barrett's esophagus, respectively. The gradient-weighted class activation mapping (Grad-CAM) was used to visually interpret the classification results of the four models. In the validation set, the models' classification abilities were evaluated. Besides, to further evaluate the classification abilities of the models, the classification results of the validation set by senior and junior physicians were compared with the models'.

    Results

    ·The 4 endoscopic image classification models of Barrett's esophagus based on deep convolutional neural networks were successfully developed. The visual interpretation of the models' classification results was presented in the form of a heat map by using Grad-CAM. In the validation set, all models presented high classification accuracy (average accuracy = 0.852) and high classification precision (average precision = 0.846). Compared with the other three models, the NASNetL model had the highest classification accuracy (0.873) and the highest classification precision (0.867), and was the best model. The NASNetL model showed similar classification ability to the senior physician. Its classification accuracy was slightly lower than that of senior physician (0.881), and higher than that of junior physician (0.855). Meanwhile, it had good classification consistency with both senior physician (Kappa=0.712, P=0.000) and junior physician (Kappa=0.695, P=0.000).

    Conclusion

    ·The endoscopic image classification models of Barrett's esophagus, developed by deep convolutional neural networks transfer learning, show good classification ability.

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    Review
    Progress of interoception based on heartbeat perception
    LIANG Yuxuan, LI Chunbo
    2022, 42 (5):  660-666. 
    doi: 10.3969/j.issn.1674-8115.2022.05.015

    Abstract ( 614 )   HTML ( 162 )   PDF (1304KB) ( 1086 )  

    In some early classical emotional theories, psychologist JAMES has proposed that emotions are the perception of physiological changes in the body, i.e., physiological changes (such as rapid heartbeat) occur before emotions such as fear and tension. Later, researchers defined the human body's ability to perceive internal physiological signals (such as heartbeat, breathing and gastrointestinal peristalsis) as interoception, in which behavioral evaluation of heartbeat perception has received extensive attention. At present, it is generally accepted that interoception can be measured in three dimensions, i.e., accuracy, sensitivity and awareness. Several studies have shown that interoception is highly correlated with the intensity of emotional experience, and the changes in interoception can also be observed in the clinical groups suffering from anxiety disorder, depression disorder, schizophrenia and other mental disorders. Using functional magnetic resonance imaging and other techniques, the researchers has found that the insula may be the center of interoception, which overlaps with the active brain regions of emotional experience, providing the neuroanatomical evidence for the emotional theory. Recently, more and more studies focus on the field of interoception, but the published domestic reviews mostly concentrated on psychology without connection with clinical practice. Therefore, this paper comprehensively reviews the interoceptive dimensions, measurement methods, research techniques and clinical researches based on heartbeat perception, and prospects the research directions of interoception.

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    Research progress of the mechanism of melatonin in the pathogenesis and course of asthma
    KANG Wenhui, CHEN Yiting, ZHAO Anda, LI Rong, LI Shenghui
    2022, 42 (5):  667-672. 
    doi: 10.3969/j.issn.1674-8115.2022.05.016

    Abstract ( 678 )   HTML ( 211 )   PDF (1255KB) ( 840 )  

    Melatonin is one of the hormones secreted by the pineal gland. The efficacy of melatonin in regulating circadian rhythm and improving sleep has been widely explored and confirmed. Studies have shown that excessive secretion or disordered melatonin can lead to adverse health effects. In recent years, accumulated evidence shows that melatonin plays an important role in anti-inflammatory, anti-cancer, immune regulation, and so on. In addition, melatonin can not only be synthesized in the human body, but also be easily obtained through external sources. Therefore, its impact on human body has attracted much attention. Based on the involvement of melatonin in airway inflammation and airway remodeling, this study systematically searched and screened 37 relevant literatures up to February 2022 and analyzed the effect of melatonin on asthma and its related mechanisms. The results showed that, on one hand, melatonin could affect gene transcription and translation by inhibiting phosphorylation signaling pathway and blocking the activation of nuclear factor-κB (NF-κB), thereby changing the expression of respiratory cytokines and inflammatory cells, reducing the generation and recruitment of eosinophils and other inflammatory cells, inhibiting cytokines and enzymes such as interleukin-1β (IL-1β), IL-4, IL-6 and caspase-1, reducing the production of mucus, playing a role in inhibiting airway inflammation, and also antagonizing the exogenous exacerbation and atopic process of asthma. On the other hand, melatonin can inhibit airway remodeling by acting on matrix metalloproteinase-9 (MMP-9), reducing collagen deposition, inhibiting goblet cell proliferation and improving smooth muscle cell phenotype. All these findings suggest that melatonin has the potential to regulate the pathogenesis of asthma by inhibiting airway inflammation and remodeling. Further exploration on the immunomodulatory function of melatonin is expected to provide new ideas for the prevention and treatment of asthma.

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    Research advances in screening modalities for colorectal cancer and colorectal adenoma
    ZHAO Min, CHU Yimin, PENG Haixia
    2022, 42 (5):  673-679. 
    doi: 10.3969/j.issn.1674-8115.2022.05.017

    Abstract ( 564 )   HTML ( 126 )   PDF (1297KB) ( 353 )  

    In recent years, the incidence and mortality of colorectal cancer (CRC) in China are on the rise, which seriously threatens the life and health of Chinese residents and causes a huge social burden. Screening to detect early colorectal cancer or adenoma (precancerous lesions) can effectively reduce the incidence and mortality of CRC. Therefore, the role and significance of screening in the prevention and treatment of CRC is significant. There are 4 types of screening techniques for CRC and colorectal adenoma. ① Stool-based tests: fecal occult based test (FOBT), fecal DNA test, fecal flora marker test, and fecal M2 pyruvate kinase (M2-PK) test. ② Imaging tests: computed tomo-graphic colonography (CTC), double-contrast barium enema (DCBE), and colon capsule endoscopy (CCE). ③ Endoscopy: flexible sigmoidoscopy (FS) and colonoscopy (CS). ④ Liquid biopsies: circulating tumor cells, circulating tumor DNA (ctDNA), circulating tumor RNAs (ctRNAs), protein markers, extracellular vesicles (EVs), etc. Among them, biomarkers DNA and RNA molecules can convey rich information about human health status and have high sensitivity and specificity in screening CRC, but they have not been widely carried out in screening adenomas, and more in-depth large-scale randomized studies of relevant biomarkers are needed. The review of CRC and colorectal adenoma screening technologies is present, introducing their principles, characteristics, and recent research advances to provide a theoretical basis for the clinical application of CRC and colorectal adenoma screening technologies.

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    Research progress in acquired cystic disease-associated renal cell carcinoma
    XIANG Han, XIANG Wu, ZHANG Weiguo
    2022, 42 (5):  680-684. 
    doi: 10.3969/j.issn.1674-8115.2022.05.018

    Abstract ( 479 )   HTML ( 98 )   PDF (1223KB) ( 267 )  

    Acquired cystic disease-associated renal cell carcinoma (ACD-RCC) is the most common type of RCC seen in patients with end-stage renal disease and acquired cystic kidney disease, and was added to the list of RCC subtypes in the 2016 World Health Organization classification. ACD-RCC is generally believed to be indolent, however, some tumors may show local recurrence and distant metastasis. Periodic screening should be performed in dialysis patients, especially in younger patients with minor comorbidities, early detection and early intervention can improve the prognosis and quality of life. Literatures published at home and abroad mainly focused on it's clinicopathological features, there is scarce research describing the imaging appearance, prognosis and surveillance of ACD-RCC. This paper summarizes the epidemiology, etiology, pathological features, imaging manifestations, treatment, prognosis and surveillance of ACD-RCC, and aims to provide a compressive understanding of the disease for clinicians.

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    Research progress of fractional CO2 laser therapy in female stress urinary incontinence
    DENG Lu, LI Jiayi
    2022, 42 (5):  685-689. 
    doi: 10.3969/j.issn.1674-8115.2022.05.019

    Abstract ( 648 )   HTML ( 121 )   PDF (1235KB) ( 617 )  

    Stress urinary incontinence (SUI) is common in middle-aged and elderly women, and its incidence is also rising with the growth of aging population. Clinically, the main first-line treatment for mild to moderate SUI is pelvic floor muscle training, but it requires a lot of time and attention from both doctors and patients. In recent years, more and more studies have been conducted to explore the laser treatment of SUI, and the feasibility of transvaginal fractional CO2 laser treatment for SUI has been preliminarily confirmed. Fractional CO2 laser can rapidly promote the proliferation of elastic fiber and collagen in the fractional photothermal area, and increase the thickness and elasticity of urethra supporting structure, thus increasing the stability of urethra and improving control of urination. However, most clinical studies in this field are non-randomized trials with small samples, and it is not yet possible to draw effective conclusions on its efficacy and safety. This paper reviews the specific mechanism and clinical research progress of fractional CO2 laser in the treatment of female SUI, and provides theoretical basis and clinical evidence reference for the application of this therapy.

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    Brief original article
    Comparison of the central positioning ability of three single-file nickel-titanium instruments in simulated curved root canals
    LI Tingting, PAN Yueping
    2022, 42 (5):  690-694. 
    doi: 10.3969/j.issn.1674-8115.2022.05.020

    Abstract ( 545 )   HTML ( 1281 )   PDF (1741KB) ( 529 )  
    Objective

    ·To compare the ability of VDW Reciproc Blue (Blue), FKG XP-endo shaper (XP) and M3-L platinum (M3) instruments to maintain the original curvature of root canal (i.e. central positioning ability) in preparing curved root canals.

    Methods

    ·Thirty resin-simulated curved single root canals were randomly divided into three groups. Root canal preparation was performed with a Blue file, an XP file or an M3 file randomly. Red or blue-black ink was injected into the root canal before and after preparation and photographs were taken under the stomatology surgical microscope. The images before and after preparation were overlapped. The apex of the prepared root was set as a circle center and the radius started at 1 mm. A concentric circle was made every 1 mm increased and sequentially incremented to 10 mm so that it intersected the image to form 10 measurement sites, and then the removal of ten measurement sites on the inside and outside of the root canal was recorded for statistical analysis. After the root canal was prepared the file was observed for tensile deformation under the stomatology surgical microscope.

    Results

    ·There was significant difference in the absolute value of the difference between the internal and external removal of root canal between the M3 group and the Blue group, as well as the M3 group and the XP group (P=0.003 1, P=0.002 0); but there was no significant difference between the Blue group and the XP group (P=0.350). The file body of XP group was observed to have de-helix deformation under the stomatology surgical microscope, which did not return to its original shape when placed in hot water, and no deformation was observed in the file body of M3 group and Blue group.

    Conclusion

    ·The three kinds of single-file nickel-titanium instruments have good formation ability when preparing the curved root canals. But XP file and Blue file can maintain the original curvature of root canals better than M3 file. XP files are prone to deformation when preparing resin root canal, therefore the use of XP should be appropriately reduced clinically.

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