Aim To construct 3 D structure model of cardiac Cav1.2 channel and check its accuracy and re-liability.Methods Homology model of Cav1.2 chan-nel α1 subunit was constructed using SWISS-MODEL server.The model was submitted to an online testing server built by University of California and scored by it.The binding of Cav1.2 channel with blocker or drug was simulated by MOE software molecular docking pro-gram to check the model′s accuracy and reliability.Re-sults Both the target sequence Cav1.2 α1 C and the template sequence Cav1.1 α1 S searched by SWISS-MODEL server belonged to L-type Ca2+channel.Since the homology was 7 1.5% revealed by sequence align-ment,homology modeling was performed using automa-ted mode.L-type Ca2+ channel blockers Verapamil, Nifedipine and Diltiazem could bind to the 3 D structure model of Cav1.2 channel,while sodium channel bloc-ker TTX could not.Furthermore,active ingredient of traditional Chinese drug Praeruptorin A and Berberine could also bind to the 3D structure model of Cav1.2 channel.Conclusion The 3 D structure model of Cav1.2 channel was constructed successfully,which provides reliable materials for further studies and estab-lishes the foundation for the application of homology modeling in the study of 3 D structure prediction of ion channels.

Objective To assess the variation of sexual hormone and mechanisms of low testosterone in young male obesity with acanthosis nigricans. Methods Retrospective analysis was performed in 125 male obesity patients [ body mass index( BMI)≥28 kg/m2 ] . According to their clinical characteristics, they were divided into two groups including obesity without acanthosis nigricans(OB group, n=62) and obesity with acanthosis nigricans(AN group, n=63). 60 normal weight men were also recruited as a control group. Body fat and body weight were measured. Blood insulin, lipid profile, sex hormones levels, and inflammation factors were measured. Parameters of each group were compared and the correlations between total testosterone level and other index were analyzed. Results All the male obesities have the significant lower total testosterone levels than those of control group(P>0. 05), and those in AN group were lower than those in OB group(P>0. 05). The BMI and body fat in OB group and AN group were both significantly higher than those in control group(P>0. 05). The fasting insulin levels in all obese men were significantly higher than those in control group(P>0. 05), highest in AN group. Triglycerides(TG) in both OB and AN group were higher than those in controls, and not significant between later 2 groups. But high-density lipoprotein-cholesterol ( HDL-C) in the two groups were significantly lower than control, which in AN group were significantly lower than OB group. Total testosterone levels in AN group were negatively correlated with weight, waist circumference, hip circumference, fasting insulin, and homeostasis model assessment for insulin resistance ( HOMA-IR ) , and also negatively correlated with inflammation factors including C-reactive protein ( CRP ) , erythrocyte sedimentation rate ( ESR) , tumor necrosis factor-α( TNF-α) , and uric acid. However, total testosterone levels in AN group were not correlated with lipid metabolism index. Conclusion Young male obesity with acanthosis are associated with secondary hypogonadism. Hyperinsulinemia, insulin resistance, and inflammatory factors are risk factors for the occurrence of this secondary male hypogonadism.

Objective:To compare the effect of hybrid open-endovascular repair (HOER) and Viabahn open revascularization technique (VORTEC)+HOER in the treatment of thoracoabdominal aortic aneurysms (TAAA).Methods:From Apr 2005 to Jul 2019, 33 TAAA patients underwent HOER including 21 cases of standard HOER, and 12 of VORTEC+HOER. The intraoperative renal ischemia time (RIT), incidence of postoperative acute kidney injury (AKI), rate of renal artery patency (RAP) and another short-term outcome were observed.Results:RIT was significantly shorter in the VORTEC+HOER group than in the standard treatment group [(9±3) minutes vs. (15±6) minutes, P<0.05]. The increase in serum creatinine (SCr) levels on the 1st postoperative day in the standard treatment group was significantly higher than that in the VORTEC+HOER group [(1.68±0.79) μmol/L vs. (1.05±0.06) μmol/L, P<0.05]. AKI occurred in 5 patients in the standard HOER treatment group (5/21, 24%), while no patient experienced AKI in the VORTEC+HOER group (0/12, 0). Conclusion:VORTEC significantly reduces RIT and postoperative SCr increasing, thereby potentially decreasing the incidence of postoperative AKI.

Objective To construct a CaME141G fusion protein-expressing plasmid,and to express,purify,and identify the activity of the recombinant protein. Methods The 141st site of the wild type CaM,E (GAG),was mutated to G (GGG),using site-specific mutagenesis technology. Escherichia coli BL-21 was transformed with the mutant plasmid. The GST-CaME141G fusion protein was mass-cultured and induced for expression. Subsequently,the GST-CaME141G fusion protein was purified using GS-4B beads. PreScission protease was applied to remove the GST,the Bradford method used to determine the concentration of purified protein,and SDS-PAGE used to detect its relative molecular weight and purity. The GST pull-down assay was used to study the protein's biological activity. Results The CaME141G protein was successfully purified at a high concentration and purity. The protein could interact with PreIQ protein fragments from the myocardial CaV1. 2 calcium channel C terminal,in a CaME141G concentration-dependent manner. Therefore,CaME141G has the ability to bind with the CaV1. 2 calcium channel. Conclusion This study successfully constructed a CaME141G fusion protein-expressing plasmid and purified the CaME141G protein. This lays a foundation for regulating the function of CaM mutations in the myocardial CaV1. 2 calcium channel,and for the study of its relationship with diseases of the cardiovascular system.

Objective:To explore the high-risk factors for parenteral nutrition associated cholestasis（PNAC）in extremely/ultra-low birth weight infants，and establish a risk Alignment Diagram prediction model.Methods:We retrospectivly analyzed the clinical data of hospitalized extremely/ultra-low birth weight infants admitted to Neonatology Department at Quanzhou Children's Hospital from January 2019 to December 2020，using multivariate Logistic regression analysis to screen for independent risk factors for the occurrence of PNAC.An Alignment Diagram model prediction model for PNAC was constructed by using R software，and the performance of the model was evaluated through receiver operating characteristic curves.Results:A total of 203 extremely/ultra-low birth weight infants were included，with a median gestational age of 29.14（28.00，30.86）weeks and a median birth weight of 1　170（1　000,1　300）g.Among them，26（12.81%）cases developed PNAC.Multivariate Logistic regression analysis showed that the duration of parenteral nutrition（ OR=1.015 ，95% CI 1.003-1.034），the cumulative amount of glucose（ OR=1.014 ，95% CI 1.001-1.028），small for gestational age（ OR=3.455 ，95% CI 1.127-10.589），and neonatal sepsis（ OR=3.142 ，95% CI 1.039-9.503）were independent risk factors for PNAC（ P＜0.05）；The four independent risk factors mentioned above were introduced into R software to construct an Alignment Diagram model，the area under the receiver operating characteristic curve was 0.835（95% CI 0.842-0.731），and the results of the Hosmer Limeshow goodness of fit test show that：χ 2=5.34，degree of freedom=8， P=0.72.A calibration curve indicated good consistency between the predicted probability of the model and the actual occurrence rate，with good accuracy. Conclusion:The Alignment Diagram model constructed based on four independent risk factors of the duration of parenteral nutrition，glucose accumulation，small for gestational age infants，and neonatal sepsis exhibits high predictive ability，and is expected to provide an intuitive and convenient visualization tool for preventing or reducing the occurrence of PNAC in extremely/ultra-low birth weight infants

Purpose: Patients with human epidermal growth factor receptor 2 (HER2)–low advanced breast cancer can benefit from trastuzumab deruxtecan. Given the unclear prognostic characteristics of HER2-low breast cancer, we investigated the prognostic characteristics of HER2-low expression from primary tumor to residual disease after neoadjuvant chemotherapy (NACT). Materials and Methods: The data of HER2-negative patients receiving NACT at our center were collected. Pathological complete response (pCR) rate were compared between HER2-0 and HER2-low patients. The evolution of HER2 expression from primary tumor to residual disease and its impact on disease-free survival (DFS) were examined. Results: Of the 690 patients, 494 patients had HER2-low status, of which 72.3% were hormone receptor (HR)–positive (p < 0.001). The pCR rates of HER2-low and HER2-0 patients (14.2% vs. 23.0%) showed no difference in multivariate analysis regardless of HR status. No association was observed between DFS and HER2 status. Of the 564 non-pCR patients, 57 (10.1%) changed to HER2-positive, and 64 of the 150 patients (42.7%) with HER2-0 tumors changed to HER2-low. HER2-low (p=0.004) and HR-positive (p=0.010) tumors before NACT were prone to HER2 gain. HER2 gain patients had a better DFS compared with HER2-negative maintained patients (87.9% vs. 79.5%, p=0.048), and the DFS of targeted therapy group was better than that of no targeted therapy group (92.4% vs. 66.7%, p=0.016). Conclusion Although HER2-low did not affect the pCR rate and DFS, significant evolution of HER2-low expression after NACT creates opportunities for targeted therapy including trastuzumab.

Objective:To study whether curcumin inhibits the proliferation and promotes the apoptosis of nephroblastoma through activating the miR-192-5p/PI3K/Akt signaling pathway.Methods:CCK-8 assay was used to investigate the effects of curcumin on the proliferation of nephroblastoma SK-NEP-1 cells and the appropriate concentration. The apoptosis rate of SK-NEP-1 cells was detected by V-FITC/PI. Luciferase reporter assay was used to verify the binding activity between miR-192-5p and PI3K. RT-PCR was performed to detect the expression of miR-192-5p at mRNA level. Western blot was used to detect the expression of PI3K and Akt at protein level.Results:Curcumin could significantly inhibit the proliferation of SK-NEP-1 cells and induce cell apoptosis in a dose-dependent manner. RT-PCR results showed that curcumin could significantly increase the expression of miR-192-5p. In addition, miR-192-5p significantly inhibited cell proliferation, induced cell apoptosis, and enhanced the effects of curcumin on the proliferation and apoptosis of SK-NEP-1 cells. Luciferase reporter assay suggested that miR-192-5p could bind to PI3K. Western blot results showed that curcumin down-regulated the expression of PI3K and Akt at protein level by mediating the expression of miR-192-5p.Conclusions:Curcumin could inhibit the proliferation and induce the apoptosis of nephroblastoma cells through mediating the expression of miR-192-5p and further inhibiting the downstream PI3K/Akt signaling pathway.

Gaining a better understanding of autoprotection against drug-induced liver injury(DILI)may provide new strategies for its prevention and therapy.However,little is known about the underlying mechanisms of this phenomenon.We used single-cell RNA sequencing to characterize the dynamics and functions of hepatic non-parenchymal cells(NPCs)in autoprotection against DILI,using acetaminophen(APAP)as a model drug.Autoprotection was modeled through pretreatment with a mildly hepatotoxic dose of APAP in mice,followed by a higher dose in a secondary challenge.NPC subsets and dynamic changes were identified in the APAP(hepatotoxicity-sensitive)and APAP-resistant(hepatotoxicity-resistant)groups.A chemokine(C-C motif)ligand 2+endothelial cell subset almost disappeared in the APAP-resistant group,and an R-spondin 3+endothelial cell subset promoted hepatocyte proliferation and played an important role in APAP autoprotection.Moreover,the dendritic cell subset DC-3 may protect the liver from APAP hepatotoxicity by inducing low reactivity and suppressing the autoimmune response and occurrence of inflammation.DC-3 cells also promoted angiogenesis through crosstalk with endothelial cells via vascular endothelial growth factor-associated ligand-receptor pairs and facilitated liver tissue repair in the APAP-resistant group.In addition,the natural killer cell subsets NK-3 and NK-4 and the Sca-1-CD62L+natural killer T cell subset may promote autoprotection through interferon-y-dependent pathways.Furthermore,macrophage and neutrophil subpopulations with anti-inflammatory phenotypes promoted tolerance to APAP hepatotoxicity.Overall,this study reveals the dynamics of NPCs in the resistance to APAP hepatotoxicity and provides novel insights into the mechanism of autoprotection against DILI at a high resolution.

OBJECTIVETo assess the effects of laparoscopic sleeve gastrectomy (LSG) on insulin secretion mode and metabolism of glucose and lipid in morbidly obese patients.

METHODSClinical data of 65 morbidly obese patients [body mass index (BMI) ≥30 kg/m] undergoing LSG at Shanghai 10th People's Hospital from August 2012 to December 2016 were retrospectively analyzed. According to the result of OGTT, these obese patients were divided into three groups: normal glucose tolerance (NGT, 23 cases), impaired glucose tolerance (IGT, 22 cases) and type 2 diabetes mellitus (DM, 20 cases) groups. Twenty-two healthy people [BMI (23.1±1.4) kg/m] were used as control group. The anthropometries parameters [weight, BMI, waist circumference, body fat percentage, excess weight loss(%EWL)], glucose metabolic indices [fasting plasma glucose (FPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index (HOMA-IR)], lipid profile (TC, TG, HDL-C, LDL-C) and inflammatory factor (UA, TNF-α) of 3 groups were detected before operation and at postoperative 1-, 3-, 6-month. These variables were analyzed among morbidly obese groups before and after surgery and compared to control group. Clinical registration number of this study was ChiCTROCSl2002381.

RESULTSBody weight, waist circumference and BMI of morbidly obese patients all decreased at postoperative 1-, 3-, 6-month. Postoperative %EWL increased obviously to (71.5±24.7)% with the highest range in DM group. Percentage of successful weight loss (%EWL>50%) in NGT, IGT and DM groups was 63.6%, 83.9% and 90.0% at postoperative 6-month respectively, and DM group was also the highest. At postoperative 6-month, HbA1c of 3 morbidly obese groups became normal; FPG and postprandial 2-hour glucose of IGT and DM group decreased to normal level; insulin level of 3 morbidly obese groups decreased obviously compared to pre-operation (all P<0.05), especially FINS and postprandial 2-hour insulin became normal without significant difference of control group (P>0.05), while postprandial 30-minute and 60-minute insulin levels in 3 groups were still higher as compared to control group. The insulin secretion curves of morbidly obese groups showed hyperinsulinemia before surgery. The peak of insulin secretion curve in IGT and DM group moved back to postprandial 120-minute before operation, and returned to 60-minute after operation, with basic normal rhythm of secretion curve. Preoperative HOMA-IR in all 3 morbidly obese groups was higher than that in control group (all P<0.05) and remarkably lower at postoperative 6-month compared to pre-operation(P<0.05). In 3 morbidly obese groups after operation, TG decreased, HDL-C increased, UA and TNF-α decreased significantly compared to before operation (all P<0.05). At postoperative 6-month, the HOMA-IR of DM group was positively correlated with BMI (r=0.236, P=0.004) and TNF-α (r=0.228, P=0.033), and was not correlated with HDL-C(P>0.05).

CONCLUSIONSLSG can effectively ameliorate hyperinsulinemia and insulin secretion curve, and improve metabolic disorder and insulin resistance of different stage in obesity patients with glucose metabolic disorder. Insulin resistance is correlated with body weight and inflammatory factors.

Objective: To evaluate the safety and efficacy of foam sclerotherapy for bleeding internal hemorrhoids. Methods: Consecutive adult patients (aged 18-60 years) with bleeding hemorrhoids were prospectively enrolled between March 2017 and March 2018. All patients were randomly assigned to the foam sclerotherapy (FS) group and the liquid sclerotherapy (LS) group. The foam and liquid sclerotherapy of hemorrhoids nucleus was performed by cap-assisted colonic endoscopy. Results: A total of 108 patients were enrolled, including 62 males (57.4%) with mean age of 44.0±7.6 years. There was no significant difference in gender, age, or body mass index between the FS (n = 55) group and the LS (n =53) group. The total amount of lauromacrogol injection in the FS group was 3.7±0.9 mL, which was significantly lower than that in the LS group (8.0±2.2 mL, P<0.001). In the LS group, 8 patients (15.1%) developed bleeding after needle removal, which was significantly higher than that in the FS group [5.5%(3/53), P=0.022]. Significant pain was found in 2 patients (3.8%) in the LS group and 0 in the FS group (P= 0.025). The short-term cure rate and effective rate of the FS group were 87.3% (48/55) and 9.1% (5/55), while those were 69.8% (37/53) and 22.6% (12/53) in the LS group, respectively (P=0.046). Conclusion Foam sclerotherapy is safe and effective in the treatment of bleeding internal hemorrhoids.