The aim of this study was to qualitatively and quantitatively detect coronavirus(CoV)in the feces of bats from Qinghua Cave,Yunnan Province,China.CoV was qualitatively tested with reverse transcription polymerase chain reaction(RT-PCR),and homology and genetic evolution were analyzed with bioinformatics software.The established reverse transcription real-time fluores-cence quantitative PCR(qRT-PCR)method was applied to CoV quantification in bat feces.The positivity rate of CoV in 306 fecal samples collected from the fulvous fruit bat(Rousettus leschenaultia)was 7.8%(24/306)according to RT-PCR.All 24 strains of CoV belonged to β-CoV,and showed a similarity of 86.8%-100.0%at the nucleotide level and 95.2%-100.0%at the amino acid level,with respect to other β-CoV sequences in the NCBI database.The positivity rate of CoV was 18.6%(57/306)according to qRT-PCR,a value higher than that according to RT-PCR(χ2=25.3,P<0.05).The mean β-CoV load was 1.3×103 copies/μL.In conclusion,the bats in Qinghua Cave,Yunnan Province,carried CoV belonging to β-CoV.The established qRT-PCR method achieved good sensitiv-ity,accuracy,reproducibility,and a higher detection rate than that of RT-PCR,and can be used for rapid detection of β-CoV in bats.

Objective:To investigate and explore the serum levels of testosterone and estradiol in correlation with disease severity and prognosis in male patients with liver failure.Methods:Sixty male cases with liver failure who received treatment from April 2022 to December 2023 were selected as the research subjects. Forty healthy subjects who underwent physical examination in the physical examination center during the same period were enrolled as the control group. The levels of sex hormones (serum testosterone and estradiol) were compared between the two groups. Logistic regression was used to analyze the diagnostic value of testosterone and estradiol for the grading of male patients with liver failure. The prognostic factors for predicting disease severity were analyzed using COX regression. The area under the ROC curve (AUC) was used to evaluate the predictive value.Results:The testosterone level was significantly higher in the healthy group than that in the liver failure group [(5.11±3.00) nmol/L vs. (2.22±2.78) nmol/L, t=4.934, P<0.001], while the estradiol level was significantly lower in the liver failure group [37.46±13.21) nmol/L vs. (113.45±67.70) nmol/L, t=-8.457, P<0.001]. Multiple discriminant logistic regression analysis results showed that estradiol and testosterone were independent predictors of the model for end-stage liver disease. Multivariate Cox regression analysis showed that testosterone was an independent prognostic factor for the 1-year mortality rate in male patients with liver failure. The area under the curve predicting the 1-year mortality rate was 0.745 after adjusting for other factors. Conclusion:Testosterone and estradiol levels are significantly altered in male patients with liver failure. Testosterone and estradiol levels in peripheral blood can effectively reflect the degree of liver function impairment and the 1-year mortality rate in male patients with liver failure, which is helpful for accurately assessing the severity of the disease and its prognosis.

The aim of this study was to qualitatively and quantitatively detect coronavirus(CoV)in the feces of bats from Qinghua Cave,Yunnan Province,China.CoV was qualitatively tested with reverse transcription polymerase chain reaction(RT-PCR),and homology and genetic evolution were analyzed with bioinformatics software.The established reverse transcription real-time fluores-cence quantitative PCR(qRT-PCR)method was applied to CoV quantification in bat feces.The positivity rate of CoV in 306 fecal samples collected from the fulvous fruit bat(Rousettus leschenaultia)was 7.8%(24/306)according to RT-PCR.All 24 strains of CoV belonged to β-CoV,and showed a similarity of 86.8%-100.0%at the nucleotide level and 95.2%-100.0%at the amino acid level,with respect to other β-CoV sequences in the NCBI database.The positivity rate of CoV was 18.6%(57/306)according to qRT-PCR,a value higher than that according to RT-PCR(χ2=25.3,P<0.05).The mean β-CoV load was 1.3×103 copies/μL.In conclusion,the bats in Qinghua Cave,Yunnan Province,carried CoV belonging to β-CoV.The established qRT-PCR method achieved good sensitiv-ity,accuracy,reproducibility,and a higher detection rate than that of RT-PCR,and can be used for rapid detection of β-CoV in bats.

Objective:To investigate and explore the serum levels of testosterone and estradiol in correlation with disease severity and prognosis in male patients with liver failure.Methods:Sixty male cases with liver failure who received treatment from April 2022 to December 2023 were selected as the research subjects. Forty healthy subjects who underwent physical examination in the physical examination center during the same period were enrolled as the control group. The levels of sex hormones (serum testosterone and estradiol) were compared between the two groups. Logistic regression was used to analyze the diagnostic value of testosterone and estradiol for the grading of male patients with liver failure. The prognostic factors for predicting disease severity were analyzed using COX regression. The area under the ROC curve (AUC) was used to evaluate the predictive value.Results:The testosterone level was significantly higher in the healthy group than that in the liver failure group [(5.11±3.00) nmol/L vs. (2.22±2.78) nmol/L, t=4.934, P<0.001], while the estradiol level was significantly lower in the liver failure group [37.46±13.21) nmol/L vs. (113.45±67.70) nmol/L, t=-8.457, P<0.001]. Multiple discriminant logistic regression analysis results showed that estradiol and testosterone were independent predictors of the model for end-stage liver disease. Multivariate Cox regression analysis showed that testosterone was an independent prognostic factor for the 1-year mortality rate in male patients with liver failure. The area under the curve predicting the 1-year mortality rate was 0.745 after adjusting for other factors. Conclusion:Testosterone and estradiol levels are significantly altered in male patients with liver failure. Testosterone and estradiol levels in peripheral blood can effectively reflect the degree of liver function impairment and the 1-year mortality rate in male patients with liver failure, which is helpful for accurately assessing the severity of the disease and its prognosis.

Objective: To research the prevalence and influencing factors of asymptomatic arrhythmia among college students, and provide scientific basis for heart rate control and health management of asymptomatic arrhythmia among college students. Methods: Using cluster sampling method, 3 012 college students who studied in the First Clinical Medical College of Henan University of Traditional Chinese Medicine and the Second Clinical Medical College of Henan University of Traditional Chinese Medicine were selected as the research objects from September to November 2022. After entering school in autumn (September in 2022), all college students were investigated by questionnaire, and the general data and living habits of asymptomatic arrhythmia college students and normal college students were compared. Logistic regression was used to analyze the influencing factors of asymptomatic arrhythmia among college students. Results: A total of 62 patients with asymptomatic arrhythmia were found. The detection rates of the patients with body mass index(BMI)≥24.0 kg/m 2 , systolic blood pressure(SBP)≥120 mmHg, diastolic blood pressure(DBP)≥80 mmHg, family history of cardiovascular disease, smoking, drinking, exercising less than three times a week, heavy study pressure, irregular work and rest, depression tendency and anxiety tendency were 5.58%, 4.49%, 5.63%, 6.18%, 4.26%, 4.50%, 3.72%, 4.29%, 4.28%, 9.15%,9.03%, which were significantly higher than those of patients with BMI<24.0 kg/m 2, SBP＜120 mmHg, DBP<80 mmHg, no family history of cardiovascular disease, no smoking, no drinking, exercise more than three times a week, little study pressure, regular work and rest, no depression tendency and no anxiety tendency (1.20%, 1.37%, 1.35%, 1.53%, 1.55 %, 1.59%, 1.27%, 1.52%, 1.38%, 1.71%,1.71%), and the differences were statistically significant ( χ 2=45.33, 25.20, 37.74, 32.24, 16.69, 17.25, 19.57, 17.83, 22.36, 37.23,39.42, P <0.01). Logistic regression analysis showed that the higher of BMI,SBP and DBP, family history of cardiovascular disease, smoking, drinking, times of exercise, heavy study pressure, irregular work and rest, depression tendency and anxiety tendency were positively correlated with asymptomatic arrhythmia of college students ( P <0.05). Conclusions The higher of BMI and blood pressure, family history of cardiovascular diseases, bad living habits and psychological status are related to asymptomatic arrhythmia in college students. It is of great significance to control weight and blood pressure reasonably and maintain good living habits and mental state for preventing and improving asymptomatic arrhythmia.

Objective:To investigate the effect and role of the hepatitis B virus (HBV) on the expression of inhibin (PHB) in the proliferation and survival of hepatocellular carcinoma (HCC) cells.Methods:The expression of PHB in 13 pairs of HBV-infected livers, normal livers and HepG2.2.15 and HepG2 cells was detected by real-time fluorescent quantitative PCR and Western blot. Liver tissues were collected from seven patients with chronic hepatitis B before and after antiviral (tenofovir) treatment, and the expression of PHB was detected by RT-PCR and Western blot. HepG2.2.15 cells were transfected with Pcmv6-AC-GFP-PHB, and control vectors were collected. DNA content was analyzed by flow cytometry. The proliferation level of each cell group was detected using the EdU cell proliferation assay. HepG2.2.15 cells transfected with Pcmv6-AC-GFP-PHB and the control vector were cultured in serum-free medium for 6 days. Apoptosis was measured at the indicated time points using fluorescence-activated cell sorting (FACS)-based Annexin-V/PI double staining.Results:Compared with normal liver tissue, the expression of PHB in HBV-infected liver tissue was down-regulated ( P < 0.01). Compared with HepG2 cells, the expression of PHB in HepG2.2.15 cells was significantly decreased ( P < 0.01). The expression level of PHB in liver tissue after antiviral treatment (tenofovir) was significantly higher than that before treatment ( P < 0.01). Compared with the control vector, the proliferation rate of HepG2.2.15 cells transfected with Pcmv6-AC-GFP-PHB was significantly lower than that of the control vector, and the apoptosis rate of HepG2.2.15 cells transfected with the Pcmv6-AC-GFP-PHB vector was significantly higher than the control vector ( P < 0.01). Conclusion:HBV down-regulates the expression of inhibin to promote the proliferation and survival of hepatocellular carcinoma cells.

OBJECTIVE To qualitatively analyze the chemical constituents from Tianzhi granules and their constituents absorbed into blood， and to provide reference for elucidating pharmacodynamic material basis of Tianzhi granules. METHODS UPLC-ESI-Q-TOF-MS/MS was adopted. The analysis was performed on an Agilent Eclipse Plus C18 column with mobile phase consisted of 0.5% formic acid solution-acetonitrile （gradient elution） at the flow rate of 0.3 mL/min； the column temperature was 40 ℃ ；the injection volume was 10 μL. Mass spectrometry was applied for the qualitative analysis under positive ionization mode and ESI ion source. Data were collected with MS-DIAL4.60， and then chemical constituents of the extract from Tianzhi granule （by 0.5% methanol） were analyzed by comparing with relevant literature， SciFinder， PubChem， MassBank， TCMSP， TCM-ID and other databases. The blank serum， administered serum and Tianzhi granule extract were compared to analyze the constituents absorbed into the blood. RESULTS One hundred compounds were preliminarily identified from Tianzhi granules， including 46 flavonoids， 8 organic acids， 8 alkaloids， 6 terpenes， 6 coumarins， 2 quinones， 1 steroids， 7 glycosides and 16 others. Based on it， 10 prototype constituents absorbed into blood were identified preliminarily， including genistein， melatonin A， chrysin-7-O- β-glucuronide， apigenin-7-O-glucuronide， wogonin， 6-O-methylbaicalin are flavonoids， 2-hydroxyquinoline and isonacolline are alkaloids， 7-hydroxycoumarin is coumarins，1-indanol is others. CONCLUSIONS 2-hydroxyquinoline， 7-hydroxycoumarin， genistein， melatonin A， isonocolline， chrysin-7-O-β-glucuronide， apigenin-7-O-glucuronide， wogonin and 6-O-methylbaicalin may be the pharmacodynamic material basis of Tianzhi granules.

This article summarizes the origins, development obstacles and job practices of International Board certified lactation consultants. This article aims at providing references for the work and practice of lactation consultants in the Chinese environment, and for managers to formulate relevant policies based on China's national conditions.

Objective: To investigate the differential expression of serum-and-glucocorticoid-inducible-kinase-2 (SGK2) in hepatocellular carcinoma (HCC) and normal liver tissues and the related mechanism mediating signal transduction of GSK-3 β / β catenin in HCC cells. Methods: Twenty pairs of matched HCC and normal tissues were collected and the situation of expression of SGK2 mRNA was detected by real-time fluorescence quantitative PCR. Western blot was used to detect the levels of SGK2 protein in human HCC cell lines (Huh-7, SMMC-7721) and normal human liver cell line (L02). SGK2 siRNA was used to transfect human HCC cell lines (SMMC-7721 and Huh-7), and then the protein expression levels of GSK-3 β/ β - catenin was successfully detected with the above-mentioned transfected cell line by western blot. Measurement data were expressed as mean ± standard deviation (±s), and the Student t -test was used as the statistical method. Results: SGK2 mRNA expression was up-regulated in all 20 HCC samples than that of the expression of matched normal liver tissues. SGK2 protein levels were significantly higher in Huh-7 and SMMC-7721 than normal human liver cell lines (P < 0.01). The downregulation of SGK2 expression in human HCC cell lines (SMMC-7721 and Huh-7) had inhibited the expression of unphosphorylated GSK-3 β. In addition, the downregulation of SGK2 expression in HCC cell lines had decreased the dephosphorylation of β - catenin to prevent degradation of the β - catenin proteasome. Conclusion SGK2 is overexpressed in HCC and mediates GSK-3β/β- catenin signaling in HCC cells.

Objective: To investigate the effect and mechanism of XTP4 gene in apoptotic hepatoblastoma HepG2 cell line. Methods: HepG2 cells were transiently transfected with small interfering RNA of XTP4 genes, plasmid pcDNA3.1/myc-His(-) A-XTP4, and hepatitis B virus X protein transactivated x gene 4 (HBX protein trans-activate gene4, XTP4) and their respective negative controls. After 48h, the overexpression and interference expression condition of XTP4 in HepG2 cells were detected by Western blot. HepG2 cells apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins P53, Bcl-2, Bax and Caspase-3 in HepG2 cells were detected by Western blot, and Bcl-2/Bax ratio was calculated. The chemiluminescence assay was used to detect activity of caspase-3 in HepG2 cells. The measured data were presented as ( ± s), and independent sample t-test was used for comparison between the two groups. Results: HepG2 cells had successfully achieved the overexpression and interference expression of XTP4 protein. Compared with the control group, the overexpression of XTP4 in HepG2 cells had significantly decreased the number of apoptotic cells (P < 0.05), and increased Bcl-2/Bax (P < 0.05) ratio, but decreased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was decreased (P < 0.05). However, interference with XTP4 expression in HepG2 cells had significantly increased the number of apoptotic cells (P < 0.05) and decreased Bcl-2/Bax (P < 0.05) ratio, but increased the expression of P53 protein (P < 0.05). The protein expression of Caspase-3 and activity of caspase-3 was increased (P<0.05). Conclusion In HepG2 apoptosis XTP4 has inhibitory effect, and its effect on inhibiting HepG2 apoptosis may be achieved by regulating the Bcl-2/Bax ratio, and the P53 protein may be involved.