Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

Objective:To investigate the safety and clinical effect of testis-sparing microsurgery(TSMS)in the treatment of benign testis tumor(BTT).Methods:We retrospectively analyzed the clinical data on 16 cases of BTT treated in the Department of Andrology of the Affiliated Hospital of Qingdao University from October 2020 to February 2023.The median age of the patients was 23 years.All the tumors were unilateral,7 in the left and 9 in the right side,with a median diameter of 1.85 cm(1.0-3.5 cm).The patients all underwent color Doppler flow imaging(CDFI),MRI,semen analysis and examination of serum T,alpha-fetoprotein(AFP),human chorionic gonadotropin(HCG)and lactate dehydrogenase(LDH),followed by TSMS.The boundaries between the tumors and normal testis tissue were accurately identified under the microscope,and the tumors and the adjacent normal testis tissue 2 mm from their margins were excised completely.Bipolar coagulation forceps were used for wound hemostasis to maximally preserve the normal testis tissue.The resected specimens were subjected to fast frozen pathology intraoperatively,and the patients were followed up for 14-40 months by regular scrotal CDFI,MRI and examinations of serum T and semen parameters.Results:The levels of serum T,AFP,HCG and LDH and semen parameters were all within the normal range preoperatively.TSMS were successfully completed in all the cases,and all were pathologically confirmed as BTT according to the latest edition of WHO Classification of Tumors:Urinary and Male Genital Tumors.CDFI showed normal blood supply within the testis tissue at 1 month after surgery.No signs of intra-testicu-lar tumor residue,recurrence or metastasis,nor significant changes in the levels of serum T,AFP,HCG or LDH or semen parameters were observed during the follow-up as compared with the baseline.Natural conception was achieved in 2 cases at 16 and 18 months re-spectively after surgery.Conclusion:BTT can be differentially diagnosed by CDFI and MRI before surgery and confirmed by histo-pathology.TSMS can achieve complete excision of the tumor,maximal sparing of the normal testis tissue and thereby effective preserva-tion of male fertility.

Objective: This article investigated the clinical characteristics and distribution of drug resistance mutation sites in HBV RT region of hepatitis B infected patients. Methods: Retrospective analysis was made on 1 948 patients with HBV infection, who had been tested for NAs resistance mutation and had a medical history of NAs in the Laboratory Department of the Fifth Medical Center of the PLA General Hospital from January 2020 to December 2021. Basic clinical information and drug resistance related mutation information were recorded. Meanwhile, the serological index data of hepatitis B were collected. Drug resistance gene mutant group and non-mutated group were grouped according to whether the drug resistance genes had a mutation in HBV RT region, and the clinical characteristics and genotype distribution of the two groups were statistically analyzed. The pattern of drug resistance gene mutation, number of mutation sites, drug resistance type and mutation of NAs resistance-related sites were analyzed in 917 patients with drug resistance gene mutation in HBV RT region. χ² Inspection was used for counting data. Meanwhile, two independent samples t-test and Wilcoxon rank sum test were used for measurement data. Results: Among the 1 948 patients with chronic HBV infection, 917 patients had drug resistance gene mutation in RT region (47.07%). The proportion of patients with acute hepatitis B and CHB in HBV RT resistance gene mutant group was lower than that in the non-mutated group, while the proportion of patients with HBV-related cirrhosis was higher than that in the non-mutated group, these differences were statistically significant. Compared with the non-mutated group in HBV RT region, the age, the positive rates of HBeAg and HBV DNA, and HBV DNA load of these patients were increased in drug resistance gene mutant group, these differences were statistically significant. Genotypes of patients in both groups were dominated by C, followed by B and D. The proportion of patients with genotype C in HBV RT drug resistance gene mutant group was higher than that of non-mutated group, the difference was statistically significant. There were 53 gene mutation patterns in 917 patients with drug resistance gene mutation in HBV RT region, and the main pattern was rtL180M+rtM204V+rtS202G (9.70%). The mutation sites were dominated by 3 (20.74%). There were 5 types of drug resistance, LAM+Ldt (21.25%) was the most. Among the 18 sites that were clearly associated with LAM, ADV, ETV and Ldt resistance in the HBV RT region, 14 sites were mutated, and the most common mutation sites were rtL180M, rtM204V, rtM204 and rtS202G. what's more, the proportion of patients with NAs drug resistance was LAM>Ldt>ETV>ADV. Conclusion: In order to prevent adverse consequences of this study such as disease recurrence or disease progression caused by HBV drug resistance, HBV infected patients, who have long-term use of NAs antiviral therapy, should monitor the level of HBV DNA and drug resistance genes in HBV RT region in order to optimize the treatment plan in time or guide individualized treatment.
Humans ; Hepatitis B virus/genetics* ; Hepatitis B, Chronic ; Antiviral Agents/therapeutic use* ; DNA, Viral/therapeutic use* ; Retrospective Studies ; Mutation ; Drug Resistance, Viral/genetics* ; Lamivudine/therapeutic use*

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.

Aim To investigate the effects of Dexmedetomidine(DEX)on HT22 cells with hypoxia/reoxygenation based on ferroptosis and the underlying mechanism.Methods HT22 cells were used to prepare H/R injury model.In order to investigate the optimal concentration of DEX, cells were divided into five groups: control group, H/R group, low concentration(H/R+DEX2.5, 2.5 μmol·L-1), medium concentration(H/R+DEX5, 5 μmol·L-1), high concentration(H/R+DEX10, 10 μmol·L-1)DEX intervention H/R group, and the survival rates of cells were detected by MTT assay.To investigate the mechanism of the protective effects on HT22 cells with H/R injury, HT22 cells were divided into four groups: control group, H/R group, H/R+DEX5 group, and H/R+DEX5 +ML385 group.The survival rates of cells were detected by MTT assay; the levels of Fe2+ were detected by FerroOrange fluorescent probe; the C11BODIPY581/591 was used to detect the change of lipid ROS on the cells; MDA and reduced glutathione kits were used to detect the content of MDA and GSH of cells respectively.The expressions of Nrf2, GPX4, TFR1 and SLC7A11 were detected by Western blot.Results Compared with control group, the survival rate of cells, the content of GSH, and the expression of Nrf2, GPX4 and SLC7A11 all significantly decreased(all P<0.05), the level of lipid ROS, the content of MDA, the level of Fe2+, and the expression of TFR1 all significantly increased in H/R group(all P<0.01).Compared with H/R group, the survival rate of cells, the content of GSH, and the expression of Nrf2, GPX4 and SLC7A11 significantly increased(all P<0.05), the level of lipid ROS, the content of MDA, the level of Fe2+, and the expression of TFR1 significantly decreased with the treatment of DEX(all P<0.05).Compared with H/R+DEX group, the survival rate of cells, the content of GSH, and the expression of Nrf2, GPX4 and SLC7A11 markedly decreased(all P<0.05), the lipid ROS, MDA and Fe2+, and the expression of TFR1 significantly increased in H/R+DEX+ML385 group(all P<0.05).Conclusions DEX can reduce H/R injury on HT22 cells by inhibiting ferroptosis, and the mechanism might be related to the promotive expression of Nrf2.

Objective To investigate the value for the level mensuration of von Willebrand factor antigen (vWF:Ag) in stroke risk assessment in the patients with non-valvular atrial fibrillation (AF).Methods 180 non-valvular AF patients were selected from the Tianjin medical university general hospital from the 2009 to 2011 for retrospective cohort study,112 males and 68 females in the group,age 61-87 years.Using the IL ACL-9000 blood coagulation instrument assay the level of vWF:Ag.Using ROC curve to analyze the diagnosis performance of vWF:Ag,using Cox regression analysis model to evaluate the of vWF:Ag effect on prognosis,using x2 test to analyze the relevance between vWF:Ag and clinical pathological factors.Compared the patients group with CHADS2 score with the patients group with CHA2DS2VASc score date using t test.Results vWF:Ag levels were control group (112 ± 34)％,paroxysmal AF group (119 ±31)％,the persistent AF group (179 ± 47)％,permanent AF group (217 ± 56)％,atrial fibrillation associated with stroke group (235 ± 104)％ respectively.There was no difference between the paroxysmal AF group and control group (q =1.75,P ＞ 0.05) ; vWF:Ag level was higher in persistent atrial fibrillation group than in paroxysmal AF group (q =10.10,P ＜ 0.01); permanent atrial fibrillation group was higher than that of the persistent AF group (q =5.21,P ＜ 0.01).The optimum cut-off point with vWF:Ag for stroke diagnosis was 188.5％,the area under ROC curve =O.843 (95％ confidence interval:0.785-0.901).In Cox regression multianalysis,the vWF:Ag (HR =0.405; 95％ CI =0.268-0.716; P =0.026),the congestive heart failure(HR =2.901 ; 95％ CI =1.837-3.951 ; P =0.001),stroke/transienl ischemic attack (HR =4.665 ; 95 ％ CI =2.837-7.291 ; P =0.000),age (HR =0.474 ; 95 ％ CI =0.211-0.765; P =0.039),the Cox analysis showed that vWF:Ag was the independent prognosis factor for stroke in AF patients.Inx2 analysis,there was the relationship between the level of the vWF:Ag and the congestive heart failure/LVdysfunction (x2 =8.227,P ＜ 0.01),hypertension (x2 =3.305,P ＜ 0.05),age (x2 =7.581,P ＜ 0.01),diabetes mellitus (x2 =6.730,P ＜ 0.01),stroke/ transient ischemic attack/thromboembolism (x2 =4.825,P ＜ 0.05),vascular disease (x2 =4.126,P ＜ 0.05).Compared the subjects with CHADS2 (score =1) with the CHA2DS2VASc(score =1),the level of the vWF:Ag was higher in patients with CHADS2 score =1 (t =4.283,P ＜ 0.01).Conclusion There was relationship between the level of vWF:Ag and main pathologic factors in patients with AF,and changed with the condition,high vWF:Ag level was an independent predictor of stroke risk,and had superior reference value for in assessment of stroke in patients with atrial fibrillation.