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.

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 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 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 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.

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 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 establish a fast,stable,and sensitive zebrafish model of osteoporosis(OP)using different method.Methods OP models were induced by iron overload or prednisolone(Pred),and bone formation and mortality were observed.The groups were divided into:Control group,model group(include FAC group and Pred group),and positive control group(AC group).Ammonium ferric citrate was used as the model drug in the iron-overload induction method.For the Pred induction models,the modeling time for the Pred-3 days post-fertilization(dpf)method was 3～9 dpf,the modeling time for the Pred-5 dpf method was 5～10 dpf,and Pred was administered from 3 dpf and removed from 7～9 dpf for the Pred withdrawal method.To compare the anti-osteoporosis(OP)effects of commonly used drugs such as Alfacalcidol(AC),Calcitriol(CA),and Alendronate(AL),it's important to select a stable and sensitive positive control drug and to further optimize different staining methods and conditions.Results There was no significant effect of ammonium ferric citrate 500 μg/mL on bone formation.Bone formation and the length of the first vertebra were significantly decreased in the Pred group induced by Pred-3 dpf compared with those in the control group(P＜0.01,P＜0.05),but zebrafish mortality was higher.There was no significant difference between the Pred-5 dpf method,but bone formation was significantly reduced in the Pred withdrawal group(P＜0.01),with no mortality.Alfacalcidol,calcitriol,and alendronate all had anti-OP effects,with CA having the most sensitive and stable anti-OP effect.Alizarin red staining showed that the optimal dye parameters were 0.02%concentration for dyeing 2 h,with washing in 0.5%KOH and glycerol under the conditions of a 3∶1 ratio for 3 h followed by a 1∶1 ratio for 14 h.The result of staining showed that calcein was more sensitive for staining bone nodes and ARS staining was more sensitive for staining the first vertebra.Conclusions The Pred withdrawal method can be used to establish a rapid,stable,and sensitive OP model in zebrafish as a reliable model for studying OP.

Objective:To retrospectively analyze the risk factors for the development of liver cancer in patients with hepatitis B-related liver cirrhosis (LC) treated and fully managed with long-term nucleos(t)ide analogues (NAs).Methods:The study subjects were derived from the follow-up cohort of chronic hepatitis B and liver cirrhosis who received antiviral therapy in the Department of Infectious Diseases of the First Affiliated Hospital of Guangxi Medical University from February 2004 to September 2019. LC patients who met the inclusion criteria were enrolled. The life-table method was used to calculate the incidence of liver cancer. Multivariable Cox regression model was used to analyze the risk factors that may affect the development of liver cancer in patients with LC. A subgroup analysis was conducted in liver cirrhotic patients who developed liver cancer to evaluate the effectiveness of antiviral treatment compliance. The 2 test was used for rate comparison. Results:The median follow-up time of 198 LC cases treated with NAs was 6.0 years (1.0-15.3 years). By the end of the visit: (1) 16.2% (32/198) of LC patients had developed liver cancer, and the cumulative incidence of liver cancer in 1, 3, 5, 7, and 9 years were 0, 8.9%, 14.3%, 18.6%, and 23.4%, respectively, with an average annual incidence of 3.1%. Among the 32 cases with liver cancer, 68.7% had developed small liver cancer (22/32). (2) Univariate Cox model analysis showed that the development of liver cancer was related to four factors, i.e., the presence or absence of LC nodules, whether the baseline was first-line medication, the family history of liver cancer, and patient compliance. The results of multivariate Cox model analysis showed that poor patient compliance and baseline non-first-line medication were risk factors for liver cancer. (3) The results of log-rank test subgroup analysis showed that the 5-year cumulative incidence of liver cancer in patients with hardened nodules was significantly higher than that of patients without hardened nodules (21.7% vs. 11.5%, P = 0.029). The 5-year cumulative incidence of liver cancer in patients with non-first-line drugs was significantly higher than that of patients with first-line drugs (22.0% vs.8.2%, P = 0.003). The 5-year cumulative incidence of liver cancer in patients with poor compliance was significantly higher than that of patients with good compliance (21.3% vs. 12.7%, P = 0.014). The 5-year cumulative incidence of liver cancer in patients with a family history of liver cancer was significantly higher than that of patients without a family history of liver cancer (22.3% vs. 8.1%, P = 0.006). (4) Compared with patients with poor compliance, patients with good compliance had higher HBV DNA negative serconversion rate (98.7% vs. 87.8%, P = 0.005), and a lower virological breakthrough rate (12.1% vs. 29.3%, P = 0.007). Conclusion:The long-term NAs antiviral therapy can reduce the risk of liver cancer, but it cannot completely prevent the development of liver cancer, especially in patients with a family history of liver cancer and baseline hardened nodules (high risk of liver cancer). Furthermore, the complete management can improve patient compliance, ensure the efficacy of antiviral therapy, and reduce the risk of liver cancer development, so to achieve secondary prevention of liver cancer, i.e., early detection, diagnosis and treatment.