Objective To construct an eukaryotic expression vector of human sTNFR1 and to investigate its inhibitory effects to the bioactivity of TNF-?.Methods The total RNA was extracted from HeLa cells and used as a template to amplify human sTNFR1 gene by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were cloned into T vector and sub-cloned into vector pcDNA3.1(-),an eukaryotic expression vector.The recombinant plasmid pcDNA3.1(-)-sTNFR1 was transfected into QSG7701 cells by using lipofectamine,RT-PCR was performed to detect the expression of sTNFR1,MTT was used to observe sTNFR1 gene 's inhibitory effect on TNF-?.Results QSG7701 has a higer expression level of sTNFR1 mRNA than pcDNA3.1(-) trancfected control.The cytotoxic effect of TNF-? was inhibited to the extent of 64.8% when its concentration was 100 ?g/L.Conclusion We constructed the eukaryotic expression vector containing human sTNFR1 gene and the cytotoxicity of TNF-? is inhibited in pcDNA3.1(-)-sTNFR1/QSG7701 cells.

Objective To construct the recombinant plasmid carrying human sTNFR1 cDNA, and express sTNFR1 in E. Coli JM109. Methods Total RNA was extracted from Hela cells, and used as a template to amplify human sTNFR1 cDNA by RT-PCR. The PCR products were cloned into T vector, and then sTNFR1 cDNA fragment was subcloned into a prokaryotic expression plasmid pMAL-c2x. The recombinant plasmid was transferred into E. Coli JM109, and induced by IPTG to express fusion protein sTNFR1-MBP. sTNFR1-MBP was purified by amylose resin affinity chromatography(ARAC), and analyzed by SDS-PAGE. Results A 558 bp human sTNFR1 cDNA was amplified by RT-PCR, and successfully inserted into plasmid pMAL-c2x. sTNFR1-MBP was produced in E.Coli after IPTG induction, and a 66 KD sTNFR1-MBP was purified by ARAC. [WTHZ]Conclusion Recombinant plasmid carrying human sTNFR1 cDNA was successfully constructed and epxressed in E. Coli JM109.

Objective To analyze the factors which influence the treatment compliance of hypertensive patients in health management. Methods Data of 6 325 hypertensive patients who received physical examination in our department were collected; 4 132 male cases and 2 193 female were included, their ages ranged from 28 to 84 years old;the average age was 61.2 ± 5.8 years. The patients of the health intervention group were randomly divided into 3 groups (group 1, group 2 and group 3). They were provided with regular health management (including weight management, catering management, sports management, medication management and monitoring of blood pressure), respectively, given different frequency of telephone follow-up (1 time per 2 months, 1 time per month, and 1 time per month), evaluating treatment compliance. All the results were analyzed and compared respectively according to the level of education, age and mental status. Data of 1 892 hypertensive patients who received outpatient services were enrolled as the control group. Among them, 4 132 were male and 2 193 were female, aged 28-84 years old, average (61.2 ± 5.8) years old. They received the traditional outpatient follow-up (outpatient service review and health education), their treatment compliance, timely correcting unhealthy lifestyle and medication method and self-testing blood pressure were evaluated. ANOVA and chi square test were used to analyze the treatment compliances and blood pressure control rates of the two groups. Result Compared with the control group, health intervention for hypertension patients could significantly improve the treatment compliance and blood pressure control rate(64.8%vs. 41.3%, 56.7%vs. 29.6%;χ2=2.827,1.382;P=0.032,0.007). Comparing the results of telephone follow-up frequency, the treatment compliance and blood pressure control rate of the 3 intervention groups were higher than those of the two other groups(77.3%vs. 65.4%, 51.7%,χ2=3.414,P=0.041;69.6% vs. 57.3%, 43.2%,χ2=2.763,P=0.028). The treatment compliance of patients with high education level was significantly higher than that of patients with low education level(68.7% vs. 59.1%, 46.4%,χ2=3.257,P=0.037;60.1%vs. 47.2%,32.8%,χ2=1.234,P=0.009). And the treatment compliance of patients with good mental state was significantly higher than that of patients with anxiety(Intervention group1:64.3%vs. 55.1%,41.9%,31.0%,χ2=2.257, P=0.016;59.4%vs.46.1%,20.9%,21.8%,χ2=3.34 5 P=0.021;Intervention group2:75.5%vs. 64.3%,51.8%,41.2%,χ2=2.932, P=0.030;68.3%vs.57.1%,39.2%, 32.1%,χ2=2.382, P=0.032;Intervention group3:86.5% vs.73.2%,62.6%,52.4%,χ2=2.435, P=0.026;75.2% vs. 68.0%,51.7%,43.3%,χ2=3.251, P=0.036). Conclusion More frequently follow-up can improve the treatment compliance and control rate of blood pressure in hypertensive patients;education, age and psychological condition are factors influencing treatment compliance in hypertensive patients.

Objective To investigate the relationship between infection of Helicobacter pylori (HP) and early renal damage in type 2 diabetes.Methods Materials of 223 customer cases who received physical examination in our department were collected and analyzed.They were divided into two groups:HP positive group and HP negative group according to the HP infection.The basic information,blood lipid,lipoprotein,albuminuria of 24 hours,fibrinogen,C-reactive protein,erythrocyte sedimentation rate,and blood urea nitrogen and creatinine were compared between two groups,respectively.Results There was no significant difference in basic information,blood glucose,glycosylated hemoglobin and blood lipid (P ＞ 0.05).The results of lipoprotein,albuminuria of 24 hours,fibrinogen,C-reactive protein,and erythrocyte sedimentation rate in HP positive group were higher than those in HP negative group (P ＜ 0.05).Conclusions The infection of Helicobacter pylori played a part in the early renal damage of type 2 diabetes.

Objective:To analyze and summarize the clinical features of pyogenic liver abscess (PLA) patients treated in 10 years, so as to provide evidence to improve current diagnosis and corresponding treatment strategies of PLA.Methods:The clinical symptoms, signs, laboratory and imaging findings, etiology and different treatments as well as corresponding efficacy and prognosis of PLA patients treated in Xiangya Hospital of Central South University during January 2010 to December 2019 were retrospectively analyzed.The chi-square was used to evaluate the differences of the categorical variables.Results:A total of 528 PLA cases were included in this study.Diabetes mellitus (46.8%, 247/528), biliary tract diseases (34.7%, 183/528) and previous hepatobiliary surgery history (19.7%, 104/528) were three most common risk factors leading to PLA. Fever (80.9%, 427/528), loss of appetite (53.6%, 283/528) and abdominal pain (51.9%, 274/528) were most common clinical manifestations. The accuracies of ultrasound, computed tomography (CT), and magnetic resonance imaging (MRI) were 95.5%(472/494), 97.0%(384/396), and 96.8%(92/95), respectively. The positive rate of pus culture (70.9%, 168/237) was higher than that of blood culture (20.0%, 42/210), and the difference was statistically significant ( χ2=115.746, P<0.01). The top three most common bacteria were Klebsiella pneumoniae (122 strains), Escherichia coli (29 strains) and Enterococcus faecium (10 strains). The drug resistance rate of Klebsiella pneumoniae to ampicillin was as high as 97.4%(112/115). The treatment programs included 200 cases of sole antibiotic treatments. Based on the antibiotic treatment, imaging guided percutaneous puncture therapy was the most commonly used treatment (45.6%, 241/528), included imaging guided percutaneous puncture and drainage (29.0%, 153/528) and imaging guided percutaneous catheterization (16.7%, 88/528). The surgical treatments included surgical catheter drainage (5.5%, 29/528) and surgical hepatectomy(11.0%, 58/528). The total number of cured cases was 495, and the cure rate was 93.8%. Conclusions:The common clinical manifestations of PLA are fever, loss of appetite, abdominal pain. Ultrasound is a routine and effective method to detect liver abscess. Klebsiella pneumoniae is the most common pathogen. Imaging guided percutaneous puncture is the most common treatment.

Objeetive To explore the relationship between metabolic syndrome and benign prostatic hyperplasia in health management.Methods A total of 5 721 patients of prostate hyperplasia was collected and divided into two groups according to metabolic syndrome.The height, weight, body mass index (BMI), and arterial blood pressure were measured.The serum PSA, cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), fasting blood glucose (FBG), postprandial blood glucose, uric acid (UA) prostate specific antigen (PSA), and free prostate specific antigen (fPSA) were monitored.The volume of prostate was measured with ultrasound.The difference was analyzed between two groups of metabolic syndrome components and the volume of prostate.The correlation between various indicators of metabolic syndrome and prostate hyperplasia and prostate volume were analyzed.Health intervention was made to observe the changes of prostate volume and symptoms in the patients with metabolic syndrome for one year.Results BMI, TG, TC, BG, prostate volume, and serum PSA were higher in the patients with metabolic syndrome than in the control group.There was significant correlation between hyperlipidemia, benign prostatic hyperplasia and diagnosis of hypertension.Prostate volume was positively correlated with obesity, hypertension, high-density lipoprotein and metabolic syndrome.After health intervention, the indicators of metabolic syndrome decreased, and the symptoms of benign prostatic hyperplasia were relieved.Conclusions Reduced obesity, hypertension, low highdensity lipoprotein and the metabolic syndrome and prostate volume were positively related.Metabolic syndrome and prostatic hyperplasia exists certain relationship.The metabolic syndrome health interventions will be necessary to manage the health of prostate hyperplasia, thus delays the occurrence and development of prostate hyperplasia disease.

Objective:To investigate expression profiles of the plasma exosomal miRNAs of the chronic hepatitis B (CHB) patients with persistently normal alamine aminotransferase (PNALT) for the first time and try to find exosomal miRNAs which could reflect liver inflammation better.Methods:Five CHB patients with liver tissue inflammation grade ≥A2 of PNALT and 5 CHB patients with liver tissue inflammation grade ＜A2 of PNALT were enrolled and their blood samples were collected.The exosomes were extracted from these blood samples and measured by electron microscope to determine the extraction effect.The exosomal miRNAs were extracted and sent for high throughput sequencing,and the expression of exosomal miRNAs in the 2 groups of patients was analyzed.Results:Under the electron microscope,exosomes were small membranous vesicles with 30-100 nm in diameter.The peak value of particle size ranged from 10 to 100 nm.High throughput sequencing showed that there were 591 differentially expressed exosomal miRNAs between the 2 groups.Compared with the control group,18 exosomal miRNAs were up-regulated and 6 exosomal miRNAs were down-regulated in PNALT patients with the liver tissue inflammation grade ≥ A2.Conclusion:Exosomal miRNAs in the CHB patients with PNALT who have the different grades of liver inflammation are differently expressed.Some of the differently expressed exosomal miRNAs are expected to be sensitive biomarkers for timely assessment of liver inflammation in the CHB patients with PNALT.

OBJECTIVES: A variety of causes can lead to cholestasis, however, cholestasis caused by Graves' disease is usually overlooked clinically. Here we analyze the clinical characteristics of Graves' disease associated cholestasis so as to have a better understanding for the disease. METHODS: We retrospectively collected 13 inpatients' data who suffered from the Graves' disease associated cholestasis in the Department of Infectious Disease of Xiangya Hospital from January 2000 to December 2018. The characteristics of the patients' age, gender, liver function, thyroid function, coagulation function, the special cardiac examination, treatment, and follow-up data were analyzed. RESULTS: Thirteen patients, including 10 males and 3 females with the age range from 33 to 55 (median 43) years old presented cholestasis, pruritus, and hypermetabolic symptoms. The levels of total bilirubin (TBIL), direct bilirubin (DBIL), glutamic-pyruvic transferase, glutamic-oxaloacetic transferase, alkaline phosphosphatase, and gamma glutamyl transpeptidase were 170.4-976.7 (median 388.8) µmol/L, 93.2-418.1 (median 199.2) µmol/L, 25.1-182.1 (median 106.4) U/L, 38.2-265.7 (median 59.7) U/L, 105.3-332.0 (median 184.5) U/L, and 20.7-345.1 (median 47.6) U/L, respectively. The levels of free triiodothyronine (FT CONCLUSIONS Graves' disease can cause cholestasis, with the low incidence. The symptoms of cholestasis can be improved or even eradicated with the cure of the Graves' disease. The cholestasis may be idiopathic. For patients with cholestasis and hyperthyroidism, Graves' disease should be considered for differential diagnosis.
Adult ; Cholestasis/etiology* ; Female ; Graves Disease/complications* ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thyroid Function Tests ; Thyroxine ; Triiodothyronine

OBJECTIVES: Long-term hepatitis B virus (HBV) infection can cause recurrent inflammation in the liver, and then develop into liver fibrosis, cirrhosis, and liver cancer. The hepatic pathological change is one of the important criteria for guiding antiviral therapy in patients with chronic hepatitis B (CHB). Due to the limitations of liver biopsy, it is necessary to find valuable non-invasive indicators to evaluate the hepatic pathological changes in CHB patients and guide the antiviral therapy. This study aims to analyze the clinical characteristics of different pathological changes in CHB patients, and to explore the factors influnencing the degree of liver inflammation and fibrosis in CHB patients with normal alanine aminotransferase (ALT). METHODS: This retrospective study was conducted on 310 CHB patients. Liver biopsy was performed in all these patients. The clinical data of the patients were collected. The liver biopsy pathological results were used as the gold standard to analyze the relationship between clinical indicators and liver pathological changes. Then CHB patients with normal ALT were screened, and the independent factors influencing the degree of liver inflammation and fibrosis were explored. RESULTS: Among the 310 patients with CHB, there were 249 (80.3%) patients with significant liver inflammation [liver inflammation grade (G) ≥2] and 119 (38.4%) patients with significant liver fibrosis [liver fibrosis stage (S) ≥2]. The results of univariate analysis of total samples showed that the ALT, γ-glutamyl transferase, alkaline phosphatase, and HBV DNA were related to the significant liver pathological changes. Among the 132 CHB patients with normal ALT, the patients with liver pathology G/S≥2, G≥2, and S≥2 were 80.3% (106/132), 68.2% (90/132), and 43.2% (57/132), respectively. The results showed that the independent influencing factor of significant liver inflammation was HBV DNA>2 000 U/mL (OR=3.592, 95% CI 1.534 to 8.409), and the independent influencing factors of significant liver fibrosis were elevated alkaline phosphatase level (OR=1.022, 95% CI 1.002 to 1.043), decreased platelet count (OR=0.990, 95% CI 0.982 to 0.998), and positive in hepatitis B e antigen (HBeAg) (OR=14.845, 95% CI 4.898 to 44.995). According to the multivariate analysis, a diagnostic model for significant liver fibrosis in CHB patients with normal ALT was established, and the area under the receiver operating characteristic curve was 0.844 (95% CI 0.779 to 0.910). CONCLUSIONS The liver pathological changes should be evaluated in combination with different clinical indicators. A considerable number of CHB patients with normal ALT still have significant liver pathological changes, which need to be identified and treated with antiviral therapy in time. Among them, HBV DNA>2 000 U/mL suggests the significant liver inflammation, and the diagnostic model for significant liver fibrosis based on alkaline phosphatase, platelet count, and HBeAg can help to evaluate the degree of liver fibrosis.
Humans ; Hepatitis B, Chronic/complications* ; Hepatitis B e Antigens/therapeutic use* ; Alkaline Phosphatase ; DNA, Viral ; Retrospective Studies ; Fibrosis ; Hepatitis B virus/genetics* ; Liver Cirrhosis/etiology* ; Inflammation/drug therapy* ; Antiviral Agents/therapeutic use* ; Alanine Transaminase

Objective: To investigate the expression of microRNA-30a (miR-30a) in hepatocellular carcinoma (HCC) and related molecular mechanisms in regulating HCC cell proliferation. Methods: A total of 30 pairs of HCC and adjacent tissue samples were collected, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of forkhead-box protein A1 (FOXA1). Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the proliferation of HCC cells, luciferase reporter gene assay was performed to verify the target relationship between miR-30a and FOXA1, and MTT assay and Western blot were used to measure the proliferation of HepG2 cells and the protein expression of FOXA1 after miR-30a transfection. The t-test was used for comparison of data between two groups, and a one-way analysis of variance was used for comparison of data between multiple groups. P < 0.05 was considered statistically significant. Results: HCC tissue had significantly lower relative expression of miR-30a than adjacent tissue (1.049 ± 0.380 vs 1.982 ± 1.013, t = 3.985, P < 0.001). At 72 hours after miR-30a overexpression, there was a significant difference in the proliferative capacity of HepG2 cells between the blank control group, the miR-30a-NC group, and the miR-30a group (0.821 ± 0.006 vs 0.816 ± 0.013 vs 0.546 ± 0.020, F = 3.396, P < 0.05), suggesting that miR-30a overexpression significantly inhibited the proliferation of HepG2 cells. FOXA1 was a target gene of miR-30a and its protein expression was negatively regulated by miR-30a, and there was a significant difference in luciferase activity between wild-type and mutant FOXA1-3’UTR vectors (1.221 ± 0.024 vs 2.658 ± 0.031, F = 6.737, P < 0.05). In HepG2 cells, miR-30a overexpression significantly inhibited the protein expression of FOXA1, and there was a significant difference in the relative expression of FOXA1 between the blank control group, the miR-30a-NC group, and the miR-30a group (1.019 ± 0.016 vs 1.022 ± 0.017 vs 0.227 ± 0.021, F = 45.43, P < 0.05). Upregulating the protein expression of FOXA1 reversed the inhibitory effect of miR-30a on the proliferation of HepG2 cells, and there was a significant difference in the proliferative capacity of HepG2 cells between the miR-30a group and the miR-30a+FOXA1 group (0.524 ± 0.023 vs 0.843 ± 0.019, t = 2.507, P < 0.05). Conclusion MiR-30a exerts its inhibitory effect on the proliferation of HCC cells by negatively regulating the expression of FOXA1.