Objective To investigate the protective effect of lipsomal clodronate against hepatic injury in rats with acute necrotizing pancreatitis (ANP). Methods 48 SD rats were randomly divided into control group, ANP group and lipsomal clodronate group, respectively. The models of ANP were established by injection of sodium taurocholate into the pancreatic capsule. Lipsomal clodronate was prepared by means of thin film. Blank liposomes and clodronate-containing liposomes was injected via caudal vein in ANP group and lipsomal clodronate group, respectively. The rats were sacrificed at 2, 6 h after ANP induction, the serum levels of ALT, AST and AMS, IL-6,IL-12 were measured, and pathologic changes of liver and pancreas were observed. Results At 6 h, serum level of ALT was (73 ± 11) U/L, (257 ± 33) U/L and (184 ± 29) U/L in control group, ANP group and lipsomal clodronate group, respectively;serum levels of AST were (190 ± 32)U/L, (590 ± 70)U/L and (430±52)U/L, respectively;serum levels of AMS were (814±80)U/L, (5031 ± 471) U/L and (2843 ± 236) U/L, respectively, serum levels of IL-6 were (26.7 ± 5.7) pmol/L, (218.0 ±4.7)pmol/L and (112.3 ± 8. O) pmol/L, respectively;serum levels of IL-12 were (4. 2 ± 1.0) pmol/L,(309.5 ± 8.5) pmol/L and (153.7 ± 6.3) pmol/L. The values in ANP group and lipsomal clodronate group were significantly higher than those in control group, while the values in lipsomal clodronate group were significantly lower than those in ANP group (P < 0. 01). Pathologic changes of liver and pancreas were significantly attenuated in lipsomal clodronate group. Conclusions Intravenous liposomal clodronate could exert protective effects on the hepatic injury in rats with ANP.

Objective To investigate the protective effect of clodronate SPIO liposomes on liver injury of rats with severe acute pancreatitis(SAP)and the role of MRI in evaluating the extent of liver injury.Methods Superparamagnetic Fe3O4 nanoparticles were prepared by chemical coprecipitation.Clodronate-SPIO-containing liposomes was prepared by the thin-film method.SAP models were prepared by a uniform injection of sodium taurocholate(2 ml/kg body weight)into the subcapsular space of the pancreas.SD rats were randomly divided into control group,SAP plus SPIO group, and clodronate-SPIO-containing liposome group.Six hours after SAP models were available,T2-weighted MRI scanning(in the same plane)of the liver of rats in each group were performed.At the end of the scanning,blood samples were taken from the supcrior mesenteric vein to measure the contents of serum ALT and AST.Meanwhile, The pathological changes in the liver and pancreas were observed.Results Transmission electron microscopic examination showed that liposomes had a uniform size.No changes in the pancreas of rats in control group were noted.The pathological changes in the pancreas and liver of rats in SAP plus clodronate-SPIO-containing liposome group were significantly milder than those in SAP plus SPIO liposome group.The contents of serum ALT and AST in rats in SAP plus SPIO liposome group were significantly higher than those in control group(P＜0.01), while the contents of serum ALT and AST in rats in SAP plus clodronate-SPIO-containing group were significantly lower than those in SAP plus SPIO liposome group(P＜0.01).The MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus clodronate-SPIO-containing liposome group was significantly lower than that in control group.The significant changes in the MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus Clodronate-SPIO liposome group were noted(P＜0.01).Conclusion Clodronate-containing liposomes have protective effects against liver injury in SAP rats and SPIO can be used as a tracer for MRI examination.

In this article,the connotation of the knowledge management and services as well as the relationship between them were introduced.The specific measures about how to carry out these management and services were described.

Objective To investigate the apoptosis of Kupffer cell (KC) induced by lipsomal clodronate in rat with acute necrotizing pancreatitis (ANP). Methods Lipsomal clodronate was prepared by means of thin film, the model of ANP was established by injection of 5% sodium taurocholate of 4 ml/kg into the pancreatic capsule. The Kupffer cells were obtained from ANP rat. After exposure to different doses of lipsomal clodronate (0, 50, 100, 150 μl) , then the proliferation and apoptosis of KC was measured by MTT, flow cytometry and agarose gel electrophoresis of DNA. Results The prepared lipsomal clodronate had an average size of 100～200 nm, the spherical shape of liposome was uniform and confirmed by transmission electron microscope. When exposed to different concentration of lipsomal clodronate for 24 h, the growth suppression rate was 17. 4% , 24. 2% and 31. 1% , respectively, while the apoptosis rate of the KC was (14. 12 ±0.37)% , (18.74±0.43)% and (27.51 ±0.39)%, respectively; the difference was statistically significantly (P<0. 01) , the DNA of KC began degradation and gradually showed clear and characteristic ladder. Conclusions Lipsomal clodronate could induce apoptosis and suppress the growth of Kupffer cells in ANP rats.

Objective To investigate the protective effects of lipsomal clodronate on renal injury in rats with severe acute pancreatifis and the assessment of renal injury. Method Totally 48 rats were randomly divided into three group:normal control group (C);SAP group, in which rats were treated with pure liposomal (P);treatment group, in which SAP rats were treated with liposomal clodronate disodium(T). The SAP model of rat was induced by injection of 5 % sodium taurochohte beneath the pancreatic membrane. Rats of normal control group received isovolumetric injections of 0.9% physiological saline solution instead of sodium taurocholate. Blood samples were collected to measure AMS,BUN,Cr,IL-6 and IL-12 at 2 hors, 6 hours after SAP. At the same time, the samples of pancreatic and renal tissues were taken for observing the pathological changes. Results Compared with controlgroup, serious renal and pancreatic damages were found in group P, and the AMS, BUN, Cr levels elevated signifi-candy (P < 0.01). Compared with group P,the renal and pancreatic damages were attenuated in group T, and the levels of Cr and AMS decreased significantly (P < 0.01), and the IL-6, IL-12 were decreased at 2 hours and 6 hours (P < 0.01). The BUN decreased significantly at6 hours (P < 0.05). Conciusions Excessive release of inflammatory mediator play an important role in renal injury in SAP. Lipsomal clodronate disodium can alleviate the damage of pancreas and kidney.

Objective To develope a new Real-time quantitative PCR assay using SYBR green as fluorescence reporter, which is rapid, specific, sensitive, cheap and accurate for the detection of mycoplasma pneumoniae(MP), and evaluated its clinical application value.Methods The sequence of the 23S rRNA gene in MP type strain FH was selected as amplified regions, and specific primers were designed.Then the related plasmids were extracted as standards,and the absolute quantitative standard curve was established.The sensitivity ,specificity of the fluorescence quantitative PCR assay was compared with the nest-PCR and kit;To calculate correlation coefficient, coincidence rate and kappa coefficient, clinical samples were detected using above-mentioned methods and cultivation,respectively.Results The detection sensitivity of the new real-time PCR and nest-PCR was 10 copies of FH DNA,while the kit 100 copies.In the specificity tests,the MP sample was positive,while mycoplasma hominis and other four bacteria were all negative.We applied this real-time PCR assay ,nest-PCR, kit and cultivation to 182 clinical specimens, and the detection rates were 55.49％, 52.75％, 47.25％ and 39.01％ ,respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and nest-PCR were 89.6％ ,0.790, respectively;while those of the new method and cultivation were 83.5 ％ ,0.678, respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and the kit were 89.6％ ,0.792,respectively;and the correlation coefficient of these two methods was 0.923,P ＜ 0.001.Conclusion Compared with other methods, the new real-time PCR assay could be used to detect mycoplasma pneumoniae quickly and economically, with high sensitivity and specificity ,revealing great utility value on varied instrumentation platforms.

Objective To investigate the antimicrobial susceptibility of spectinomycin?minocycline?azithromycin and sparfloxacin to mycoplasma(Uu and Mh)and therapeutic effect of spectinomycin to my-coplasma infection in genitourinary tract.Methods①The susceptibility test:each of the4drugs was divided into two concentrations.One was at1?g/mL(sensitive concentration)and the other was at4?g/mL(resistant concentration).If mycoplasma does not grow in both concentrations,it means the drug tested is sensitive.If it grows in both concentrations,the drug tested is resistant.If mycoplasma grows in lower concentration and does not in higher concentration,it means moderate sensitive.②Treatment regimen:Spectinomycin was injected,2g/d IM,for7-10days as a course of treatmeant.Patients were followed-up7days later and2~4weeks after treatment.Results①Among1658specimens,519were found Uu positive,and61Mh positive.The resis-tance rates of Uu to4different drugs were:7.7%for minocycline,21.4%for sparfloxacin,13.9%for azithromycin and7.3%for spectinomycin.Whereas,those of Mh were:18.0%,45.9%,54.1%,and29.5%re-spectively.②The clinical effect of spectinomycin was:out of43treated patients,37(86.0%)cured,4(9.3%)markedly improved,2(4.7%)failed.Total effective rate was95.3%and so was the elimination rate of my-coplasma.Conclusion The resistant rate of mycoplasma to spectinomycin is lower than that to minocycline?azithromycin and sparfloxacin,and the former is widely used in the treatment of mycoplasma(especially Uu)infection,with a satisfactory clinical effect.

Objective:To explore the effect of formative evaluation based on Rain Classroom on clinical skills teaching.Method:s A total of 70 students in clinical medicine from Grade 2016 were enrolled and divided into experimental group ( n=35) and control group ( n=35). The experimental group adopted Rain Classroom combined with formative evaluation, while the control group used conventional teaching method and evaluation. Test scores of the two groups were compared and students' satisfaction was collected via questionnaire. T-test was performed using SPSS 17.0. Result:The total score of clinical skills test in the experimental group was (84.11±7.76), which was significantly higher than that in the control group (74.37±12.58), and the difference was statistically significant ( P<0.05). According to the questionnaire survey, the experimental group was significantly better than the control group in terms of enhancing learning interest, improving knowledge comprehension, clinical skills, clinical thinking and analytical ability, as well as better satisfaction towards teaching ( P<0.05). Conclusion:Formative evaluation based on Rain Classroom is helpful to improve the teaching effect on clinical skills and comprehensive abilities of students, so as to promote their overall development.

Objective To investigate the efficacy of entecavir (ETV) versus tenofovir disoproxil fumarate (TDF) and the treatment measures for poor response in previously untreated chronic hepatitis B (CHB) patients with high viral load. Methods A total of 165 CHB patients who received antiviral therapy and met the inclusion criteria in Department of Infectious Diseases, The First Affiliated Hospital of Guangxi Medical University, from June 2016 to July 2021 were enrolled. The patients enrolled had a baseline HBV DNA level of > 6lg copies/ml and were previously untreated CHB patients who had used ETV or TDF for 48 weeks, and quantitative real-time PCR was used to measure HBV DNA. Virologic response rate was calculated after 48 weeks of treatment; a logistic regression analysis was used to investigate the influencing factors for the response of HBV DNA < 500 copies/mL and HBV DNA < 100 copies /mL at 48 weeks; a stratified analysis was performed to compare the virologic response rate of HBV DNA < 500 copies /ml and HBV DNA < 100 copies/ml after 48 weeks between the patients with different ages, sexes, baseline HBV DNA levels, baseline alanine aminotransferase (ALT) levels, types of first-line medication, and HBeAg statuses. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups, and the binary logistic regression model was used for multivariate analysis. Results After 48 weeks of treatment, 85.5% (141/165) of the patients achieved an HBV DNA load of < 500 copies/mL, and 66.1% (109/165) of the patients achieved an HBV DNA load of < 100 copies /mL, with no significant difference in treatment outcome between the ETV group and the TDF group. The multivariate logistic regression analysis showed that sex( OR =2.793, 95% CI : 1.197-6.517), baseline HBV DNA( OR =0.369, 95% CI : 0.142-0.959), baseline ALT( OR =4.556, 95% CI : 1.770-11.732), and baseline HBeAg( OR =0.120, 95% CI : 0.033-0.429) were influencing factors for complete virologic response(all P < 0.05). For the patients with normal ALT (≤40 U/L) at baseline, 75.6% (34/45) achieved an HBV DNA load of < 500 copies/mL after 48 weeks of treatment, and 53.3% (24/45) achieved an HBV DNA load of < 100 copies/mL, with no significant difference in treatment outcome between the ETV group and the TDF group. For the patients with abnormal ALT (> 40 U/L) at baseline, 89.2% (107/120) achieved an HBV DNA load of < 500 copies/mL after 48 weeks of treatment, and the proportion of such patients in the TDF group was significantly higher than that in the ETV group (96.1% vs 84.1%, χ 2 =4.386, P =0.036); 70.8% (85/120) achieved an HBV DNA load of < 100 copies/mL, the proportion of such patients was no significant difference between the TDF group and the ETV group (78.4% vs 65.2%). The response of HBV DNA < 100 copies/ml of the normal baseline ALT group and the abnormal baseline ALT group, there were no significant differences between the patients aged≤30 years and aged > 30 years (77.8% vs 47.2%, 85.2% vs 66.7%). For the patients who did not achieve complete virologic response (HBV DNA ≥100 copies/mL) after 48 weeks of treatment, 87.9% (29/33) achieved complete virologic response after the original treatment regimen was prolonged for 48 weeks, and 100% (9/9) of the patients achieved complete virologic response after switching to or adding the first-line nucleos(t)ide analogues (NUCs) without cross-resistance sites with the original regimen for another 48 weeks. Conclusion The patients aged > 30 years should receive antiviral therapy as early as possible, regardless of viral load and ALT level, especially those with a family history of liver cirrhosis or hepatocellular carcinoma; the patients aged ≤30 years who have a normal ALT level and a high viral load should consider initiating antiviral therapy after providing informed consent. For the patients with poor response after 48 weeks of treatment, first-line NUCs without cross-resistance sites with the original regimen should be switched to or added in time.

Lipid metabolism disorders contribute to hyperlipidemia and hepatic steatosis. It is ideal to develop drugs simultaneous improving both hyperlipidemia and hepatic steatosis. Nitazoxanide is an FDA-approved oral antiprotozoal drug with excellent pharmacokinetic and safety profile. We found that nitazoxanide and its metabolite tizoxanide induced mild mitochondrial uncoupling and subsequently activated AMPK in HepG2 cells. Gavage administration of nitazoxanide inhibited high-fat diet (HFD)-induced increases of liver weight, blood and liver lipids, and ameliorated HFD-induced renal lipid accumulation in hamsters. Nitazoxanide significantly improved HFD-induced histopathologic changes of hamster livers. In the hamsters with pre-existing hyperlipidemia and hepatic steatosis, nitazoxanide also showed therapeutic effect. Gavage administration of nitazoxanide improved HFD-induced hepatic steatosis in C57BL/6J mice and western diet (WD)-induced hepatic steatosis in Apoe ^-/- mice. The present study suggests that repurposing nitazoxanide as a drug for hyperlipidemia and hepatic steatosis treatment is promising.