ObjectiveTo investigate the regulatory effect of icariin （ICA） on transforming growth factor-β₁ （TGF-β₁）/Smad pathway in bone microvascular endothelial cells （BMECs） and the effect on autophagy in BMECs. MethodsBMECs were isolated and cultured， and the cell types were identified by immunofluorescence. Cells were divided into the control group， model group （0.1 g·L^-1 methyl prednisolone）， ICA group （0.1 g·L^-1 methyl prednisolone +1×10^-5 mol·L^-1 ICA）， and TGF-β inhibitor group （0.1 g·L^-1 methyl prednisolone +1×10^-5 mol·L^-1 ICA +1×10^-5 mol·L^-1 LY2157299）. Transmission electron microscopy was used to observe the ultrastructure and autophagosome number of BMECs. Autophagy double-standard adenovirus was used to monitor the confocal autophagy flow generation of each cell. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the gene and protein expression of autophagy in the TGF-β₁/ Smad pathway. ResultsAfter cell separation culture， platelet endothelial cell adhesion molecule （CD31） and von willebrand factor （vWF） immunofluorescence identified BMECs. Transmission electron microscopy showed that the cell membrane was damaged， and the nucleus was pyknotic and broken in the model group. Compared with the model group， the ICA group had complete cell membranes， clear structures， with autophagy-lysosome sparsely distributed. The confocal photo showed that BMECs had autophagosomes and autophagy-lysosomes， and the autophagy expression of the ICA group was similar to that of the blank group. Compared with the blank group， in the model group and the LY2157299 group， autophagosomes and autophagy-lysosomes were barely seen in the autophagy flow. Compared with the blank group， the mRNA and protein expressions of autophagy effector protein 1 （Beclin1） and microtubule-associated protein 1 light chain 3B （LC3B） in the model group were significantly decreased （P<0.01）， and those of ubiquitin-binding protein （p62） were significantly increased （P<0.01）. The mRNA expression of TGF-β₁， Smad homolog 2 （Smad2）， and Smad homolog 3 （Smad3） decreased （P<0.05， P<0.01）. The protein expressions of TGF-β₁， p-Smad2， and p-Smad3 were significantly decreased （P<0.01）. Compared with those of the model group， the mRNA and protein expression of Beclin1 and LC3B in BMECs of the ICA group increased （P<0.01）， and those of p62 significantly reduced （P<0.01）. The mRNA expression of TGF-β₁， Smad2， and Smad3 increased significantly （P<0.01）. The protein expression of TGF-β₁， p-Smad2， and p-Smad3 increased significantly （P<0.01）. Compared with those in the model group， the mRNA and protein expressions of Beclin1， LC3B， and p62 in the inhibitor group were not statistically significant. The expression of key genes and proteins of the TGF-β₁ pathway in the inhibitor group was not statistically significant. ConclusionICA can promote glucocorticoid-induced autophagy expression of BMECs， and its mechanism may be related to activating the TGF-β₁/Smad signaling pathway.

A convolutional neural network-based algorithm is proposed for calculating lower limb joint angle in X-ray films.After identifying the region of interest of a specific category in X-ray films through Yolov5 object detection model,U-Net model is used to perform heat map regression for identifying the key feature points,and then the lower limb joint angle is calculated.The results show that the proposed algorithm has higher accuracy than the previous algorithms and can obtain accurate and reliable results,providing references for clinical research and practice.

Objective To explore the mechanism of Yinchenhao Decoction in preventing and treating MAFLD based on"Intestinal TPH1-hepatic HTR2A axis".Methods Twenty-four C57BL/6J mice were arbitrarily splited up into control group,model group and Yinchenhao decoction group,eight in each group.Mice in the Yinchenhao decoction group and model group were fed with high-fat diet.After 12 weeks,the Yinchenhao decoction group was given Yinchenhao decoction by gavage,once a day for 4 consecutive weeks.Histopathological changes were observed by HE staining and oil red O staining.Serum HDL-C,LDL-C,TC,TG,AST,ALT and 5-HT contents,liver TC,TG,DAG,PLC contents were detected.Intestinal TPH1,SERT and liver HTR2A,SREBP-1c,GPAT1,FASN mRNA levels;Intestinal TPH1,SERT and liver HTR2A,SREBP-1c,GPAT1,FASN,P-PI3K,PKC-ε,P-AKT,P-mTOR protein expression level were detected.Results In the control group,the hepatocytes were arranged neatly without significant steatosis;In the model group,the hepatocytes were swollen in volume with significant steatosis;Compared with the control group,hepatocyte steatosis was significantly reduced in the Yinchenhao decoction group.Compared with the control group,liver lipid deposition was significantly higher in the model group,and the Yinchenhao decoction group significantly improved liver lipid deposition.Compared with the control group,the liver TG,TC levels in the model group were significantly increased(P<0.05);the serum AST,ALT,HDL-C,LDL-C,TG,TC levels were significantly increased(P<0.05);the serum 5-HT and liver DAG,PLC was significantly increased(P<0.05);the mRNA expression levels of HTR2A,SREBP-1c,GPAT1,FASN in the liver and TPH1 in the Intestinal were significantly increased,SERT in the Intestinal were significantly decreased(P<0.05);the protein expression levels of HTR2A,SREBP-1c,GPAT1,FASN,P-PI3K,P-AKT,P-mTOR,PKC-ε in the liver were significantly increased,and TPH1 in the Intestinal were significantly increased,SERT in the Intestinal were significantly decreased(P<0.05).Compared with the model group,the liver TG,TC levels in Yinchenhao decoction group were significantly decreased(P<0.05);the serum AST,ALT,LDL-C and TG levels were significantly decreased(P<0.05);the serum 5-HT and liver DAG,PLC level was significantly decreased;The mRNA expression levels of HTR2A,SREBP-1c,GPAT1,FASN in the liver and TPH1 in the Intestinal were significantly decreased,SERT in the Intestinal were significantly increased(P<0.05);the protein expression levels of HTR2A,SREBP-1c,GPAT1,FASN,P-PI3K,P-AKT,P-mTOR,PKC-ε in the liver were significantly decreased and TPH1 in the Intestinal were significantly increased,SERT in the Intestinal were significantly increased(P<0.05).Conclusion Yinchenhao decoction may regulate liver TG synthesis through Intestinal TPH1-hepatic HTR2A axis,thereby inhibiting the occurrence and development of MAFLD.

Objective To investigate the characteristics of the gut microbiota of patients with non-alcoholic fatty liver(NAFLD)damp-heat accumulation syndrome and its correlation with serum metabolites.Methods 40 NAFLD patients with damp-heat accumulation,19 NAFLD patients with depressed liver and deficient spleen and 32 healthy people were selected,using 16 SrRNA amplicon sequencing technology and LC-MS/MS technology to test gut microbiota and serum metabolites.The correlation between gut microbiota and serum metabolites was analyzed using Spearman rank correlation.Results Compared with the healthy control group,the relative abundance of Shigella and Collinsella in the NAFLD with damp-heat accumulation group was higher,and the relative abundance of Bifidobacterium was lower,there was no difference between NAFLD with damp-heat accumulation group and depressed liver and deficient spleen group.Compared with the healthy group and NAFLD with depressed liver and deficient spleen group,the level of L-Tryptophan in NAFLD with damp-heat accumulation group was significantly higher;compared with healthy people,the level of Xanthurenic acid in NAFLD with damp-heat accumulation group increased.L-Tryptophan is negatively correlated with Agrobacterium,and Xanthurenic acid is positively correlated with Acinetobacter,Leuconostoc,and Collinsella.Compared with the healthy group and NAFLD with depressed liver and deficient spleen group,the level of L-Thyroxine in NAFLD with damp-heat accumulation group was significantly lower;compared with healthy people,the level of L-phenylalanine in NAFLD with damp-heat accumulation group was increased,and compared with NAFLD with depressed liver and deficient spleen group,its level was significant decline.L-Thyroxine is negatively correlated with Megamonas,Acinetobacter,and Subdoligranulum.Compared with the healthy control group,the levels of Glycochenodeoxycholate,Deoxycholic Acid,and Glycocholate in the NAFLD with damp-heat accumulation group were significantly higher.Compared with the NAFLD depressed liver and deficient spleen group,the above metabolites were not significantly different.Glycochenodeoxycholate is positively correlated with Collinsella and Agrobacterium,and Glycocholate is positively correlated with Acinetobacter,Leuconostoc,and Shigella.Compared with the healthy control group and NAFLD with depressed liver and deficient spleen group,the levels of Inosine 5'-Monophosphate and guanine nucleoside in NAFLD with damp-heat accumulation group were significantly increased;compared with the healthy control group,the level of uric acid was significantly increased,and there was no significant difference compared with the NAFLD with damp-heat accumulation group.Inosine 5'-Monophosphate was positively correlated with Leuconostoc,negatively correlated with Bifidobacterium,and guanosine was positively correlated with Leuconostoc.Conclusion NAFLD patients with damp-heat accumulation syndrome have gut microbiota imbalance and metabolic disorders.The gut microbiota imbalance of NAFLD with damp-heat accumulation syndrome is closely related to the host tryptophan,phenylalanine,and purine metabolism disorder.

Objective:To investigate the epidemiology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in patients with maintenance hemodialysis (MHD) in Jiangsu province during SARS-CoV-2 pandemic in China from December 7, 2022 to January 27, 2023, and to analyze the influencing factors of all-cause death.Methods:It was a multi-center cross-sectional investigation. Structured questionnaire was used to collect patient information by medical staff of each hemodialysis center (room) as investigators. Part of the demography data and laboratory examination data came from the Jiangsu Province Hemodialysis Data Information System. MHD patients from hemodialysis centers (rooms) at all levels of medical institutions and independent hemodialysis institutions in Jiangsu province during the outbreak of SARS-CoV-2 infection were included, and the clinical characteristics and all-cause mortality of confirmed and suspected cases of SARS-CoV-2 infection were analyzed.Results:Questionnaire surveys and data analysis on 57 278 patients in 407 hemodialysis centers (rooms) were completed, accounting for 90.41% of the total number of MHD patients (63 357 cases) in Jiangsu province during the same period. There were 24 038 cases (41.97%) of SARS-CoV-2 infection and 14 805 cases (25.85%) of suspected infection, which were widely distributed in all dialysis centers in Jiangsu province. After clinical classification of 38 843 confirmed and suspected SARS-CoV-2 infection cases, 3 662 cases were severe and critical cases, accounting for 9.43% of the infected and suspected cases. Among the patients who had completed the questionnaires, there were 1 812 all-cause deaths, with an all-cause mortality rate of 3.16%. Multivariate logistic regression analysis showed that elderly (taking ≤50 years as a reference, 51-59 years: OR=1.583, 95% CI 1.279-1.933, P=0.001; 60-69 years: OR=3.972, 95% CI 3.271-4.858, P<0.001; 70-79 years: OR=7.236, 95% CI 5.917-8.698, P<0.001; ≥80 years: OR=11.738, 95% CI 9.459-14.663, P<0.001), male ( OR=1.371, 95% CI 1.229-1.529, P<0.001), and co-infection with hepatitis B virus (HBV) (positive serum HBV surface antigen, OR=0.629, 95% CI 0.484-0.817, P<0.001) were independent influencing factors for all cause mortality. Receiver-operating characteristic curve analysis showed that the area under the curve for male, age and current HBV infection prediction of all-cause death was 0.529 ( P<0.001), 0.724 ( P<0.001) and 0.514 ( P=0.042), respectively, and the cut-off value for age prediction of all-cause death was 65.5 years old. Compared with patients without HBV infection, MHD patients with HBV infection significantly reduced the proportion of severe and critically ill patients, all-cause hospitalizations and all cause deaths when infected with SARS-CoV-2 (4.99% vs. 6.41%, χ2=6.136, P=0.013; 8.90% vs. 11.44%, χ2=11.662, P<0.001; 2.01% vs. 3.37%, χ2=10.713, P=0.001, respectively). Conclusion:The MHD patients in Jiangsu province are susceptible to SARS-CoV-2. Elderly age and male gender are independent risk factors for death in MHD patients during the epidemic, while the HBV infection may be a protective factor for death of MHD patients infected with SARS-CoV-2.

Gitelman syndrome is a rare disease.It is easy to be misdiagnosed and missed diagnosis due to the diverse clinical symptoms.A girl with long-term hypokalemia,who presented with intermittent pain of lower limb muscle and physical retardation,was treated in Xiangya Hospital,Central South University.Laboratory examination confirmed the severe hypokalemia and metabolic alkalosis.Gene sequencing indicated SLC12A3 gene mutation and the patient was finally diagnosed as Gitelman syndrome.Patients with chronic hypokalemia and metabolic alkalosis need to conduct gene sequencing to confirm the diagnosis.Gene therapy is expected to be the most effective treatment for this disease.

Objective To evaluate the therapeutic effect of small bone flap craniotomy decompression of posterior cranial fossa and duraplasty in the treatment of Chiari malformation type Ⅰ.Methods The clinical data of 45 Chiari malformation type Ⅰ patients who were treated with small bone flap craniotomy decompression of posterior cranial fossa and duraplasty were retrospectively analyzed,31 cases among them with syringomyelia.Results According to Tator etc.standard,1 month after surgery,the excellent in 30 cases,good in 15 cases.Follow up from 6 months to 6 years,the excellent in 37 cases,good in 8 cases.Among 31 patients with syringomyelia,26 cases were syringomyelia subsided,5 cases were not obvious change.Conclusion The small bone flap craniotomy decompression of posterior cranial fossa and duraplasty can make the craniocervical decompression,and has obvious effect of treating syringomyelia,is safe and effective in treatment of Chiari malformation type Ⅰ.

Objective To explore the responses of antigen-specific T cells stimulated by hepatitis B virus(HBV)-specific proteins in chronic hepatitis B patients accepting antiviral therapy. Methods Seventeen patients with chronic hepatitis B (CHB) accepting antiviral therapy were included in this study. The peripheral blood monocular cell ( PBMC) were separated from the whole blood collected at the three different time of before and one and three months after accepting antiviral therapy. ELISPOT assay was used to detect the frequency and strength of secreting IFN-γ cells of PBMC stimulated by HBsAg, HBcAg and HBeAg. HBV virus loading, HBsAg, HBeAg, ALT and AST in serum were detected at the same time. Results After three months therapy, ALT, TBiL were improved in all patients, and HBV DNA level were dropped and undetectable in 11 cases. The rates of T cell response in patients to HBV specific proteins were 64. 7% , 76. 5% and 82. 4% at the time of before and one and three months after accepting antiviral therapy, respectively. The frequency of responses of antigen-specific T cells stimulated by HBcAg was higher than that stimulated by HBsAg or HBeAg, and the frequency was enhanced after antiviral therapy. The average response magnitude was expressed as spot forming cells (SFC) per million input cells. SFC of T cell responses to HBcAg was also higher than to HBsAg or HBeAg. There was no significant difference in SFC of T cell responses to HBsAg or HBeAg at the time of before and after antiviral therapy, but there were significant difference in SFC of T cell responses to HBcAg at the time of before and after antiviral therapy. SFC of T cell responses to HBcAg was negatively associated with HBV DNA, and no associated with level of ALT in serum. Conclusion The responses of antigen-specific T cells were improved in CHB patients accepting antiviral therapy which associated with the decrease of HBV DNA. It suggested to investigate HBV specific T cell responses was important.

Objective: To investigate the changes of myo cardial contractile function during myocardial stunning in calcium overload rats and the protective effects of tetrandrine. Methods: Forty-six rats were randomized into control, myocardial ischemia, myocardial stunning, low and high dose of tetrandrine groups. Another 10 rats were used to identify the calcium overload. vitamin D3 (0.3 million Unit/kg) and nicotinic acid were adm inistered. After 16 d when calcium overload occured, left anterior descending ar tery was ligated. Twenty minutes of myocardial ischemia followed by 60 min of re perfusion was induced. The contractile function parameters were determined dynam ically. At the end of experiment, myocardial cytosolic ［Ca2+］i was deter mined in various groups. In tetrandrine groups, tetrandrine (62.2 or 93.6 μmol/ kg ) was administered by gastrogavage daily.After 16 d, the rats undergone the e xperiments mentioned above. Results: Sixteen days after vitamin D3 , nicotinic acid were given, ［Ca2+］i increased by 2.6 folds (146.8±10.8 ) vs (368.5±22.6) nmol/L, (P<0.01). Whereas, ［Ca2+］i in tetrand rine groups were (210.8±16.4) and (198.6±15.3) nmol/L, which were significantl y lower than that of calcium overload group. Twenty minutes of myocardial ische mia resulted in the decrease of dp/dtmax and Vmax in all groups with the most si gnificant in stunning and calcium overload groups. The contractile function rest ored gradually after reperfusion. At all time points, dp/dtmax and Vmax in both tetrandrine groups were higher than those in both stunning and calcium overload groups. And effect with higher dose of tetrandrine were more significant than in low dose of tetrandrine. After 60 min of reperfusion, dp/dtmax in stunning, cal cium overload, low and high dose of tetrandrine groups were 49.7%, 51.5%, 71.0% and 83.4% of that in control, respectively， and Vmax were 55.0%, 49.8%, 73.9% and 77.5% of that in control, respectively. Conclusion: T he myocardial contractile function in vitamin D3-induced calcium overload gro up is impaired. On basis of myocardiocyte calcium overload, transient ischemia l eads to myocardial stunning. At the stage of ischemia, the impaired degree of my ocardial contractile function is similar to that in stunning group, suggesting a t this stage the effect of ischemia on myocardial function is greater than that of calcium overload. Tetrandrine chronically improves the myocardial function in Vitamin D3-induced calcium overload rats.

Objective:To observe the effect of percutaneous tr ansluminal coronary angioplasty (PTCA) on coronary circulating tumor necrosis fa ctor-α (TNF-α) activity. Methods: Plasma TNF-α levls were measured with radioimmunoassay and bioactive assay respectively. Result s: Plasma TNF-α activity in femoral artery (AO) was significantly incr eased immediately after PTCA ［(15.86±3.75) U/ml vs （41.32±4.36） U/ml， P<0.01］, and plasma TNF-α activity in coronary sinus was remarkably incre ased immediately after PTCA　［(16.72±4.14） U/ml vs （65.61±6.25） U/ml, P<0.01］. There was no change in plasma TNF-α activity in AO 24 h after PT CA ［(18.32±5.12） U/ml vs （15.86±3.75） U/ml, P>0.05］. Conclu sion: The increase in plasma TNF-α activity after PTCA may be associat ed with the injury of coronary artery caused by PTCA, suggesting that TNF-α ma y be involved in the coronary occlusion and the development of coronary restenos is after PTCA.