Objective To explore a simple and efficient method for isolating and culturing mouse primordial germ cells (mPGCs) in vitro in order to provide the sufficient mPGCs sources of mouse model .Methods The 12 .5 dpc mouse embryonic gen-ital ridge were isolated in vitro and cut into pieces ,then the organization to adherent culture was performed in a-MEM medium con-taining fetal calf serum and insulin-transferrin-selenium (ITS) and follicle stimulating hormone(FSH) and recombinant endothelial growth factor(rEGF) .The inverted microscope was used to observe the cellular morphology .Flow cytometry was used to identify the stage specific embryonic antigens of cultured cells .Reverse transcription-polymerase chain reaction(RT-PCR) and immunofluo-rescence were used to identify the pluripotency gene Oct4 expressions of stem cells and specific genes in the meiosis phaseⅠ .Results The in vitro isolated and cultured mPGCs showed good cell morphology ,extremely strong proliferation capacity and the positive expression of SSEA-1 and negative expression of SSEA-4 ,Oct4 ,meanwhile the expressions of Stra8 ,Vasa ,Scp3 ,Zp3 were detected to be positive .Conclusion Using this culture method can culture the high purity of mPGCs with the excellent stem cell properties and extremely strong proliferative ability ,moreover which makes the cells entering the meiosis stage .

Objective To establish a method for determination of polysaccharides in Weixuening Granule, which can be used for quality control of the preparation. Methods Polysaccharides was determined by improved differential phenol-sulfuric method with glucose as reference, and the color conditions were optimized at the wavelength of 487 nm, with the concentration of phenol and sulfuric acid, and reaction time as investigated factors. Results The best color conditions of polysaccharides were 0.04 g/mL phenol 1 mL, 87.5% sulfuric acid 5 mL, and reaction time of 15 min. Glucose had a good linear relationship with absorbanceΔA at the range of 15.3-306.0μg, r=0.999 7. The average recovery was 103.5%, and RSD was 2.9%. The average content of polysaccharides in Weixuening Granule was 27.04%and RSD was 1.1%. Conclusion The method is simple, accurate, stable and reliable, and can be used to determine the content of polysaccharides in Weixuening Granule.

Aims The purpose of this study is to investigate thediagnostic value of SWEfor fibrosis in patients with CHBand the factorsinfluencing the accuracy. Methods From July 2013 to October 2015, 261 patients with CHB were recruited from the First Affiliated Hospital of Sun Yat-sen University.All patients received SWE, anthropometry measurement, blood cell count, liver function test and virological indicators measurement. Liver fibrosis was staged from F0 to F4 by METAVIR scorebased onliver biopsy results of 133 CHB patients , while 128 patients were diagnosed as decompensated cirrhosis. Diagnostic accuracyof SWE were evaluated by Receiver Operating Characteristic Curve (ROC) using liver hepatic pathology and decompensated cirrhosis as gold standards. Logistic model was used to find out confounding factors that influence the accuracy of SWE. Results The Area Under ROC (AUC) for liver stiffness measurement with SWE were 0.891, 0.932 and 0.910 for the diagnosis of significant fibrosis (≥ F2), advanced fibrosis (≥F3) and cirrhosis (F4), respectively. A multifactor logistic regression combined modelwas built and showed that hepatic steatosis will decrease the accuracy of SWE. Conclusion SWE could be a valuable method for the noninvasive liver fibrosis assessment. The accuracy of SWE may be influenced by hepatic steatosis.

Objective To investigate the value of serum concentration of VEGF-C in the prognosis of advanced pancreatic cancer. Methods Thirty-five patients with advanced pancreatic cancer were selected from Aug. 2006 to Feb. 2008, ELISA method was used to detect the serum level of VEGF-C, CA19-9 and KPS score was calculated, and survival was analyzed by Kaplan Meier method. The survival difference was calculated by log rank. Cox regression model was used to perform univariate and multivariate analysis. Results The mean serum concentration of VEGF-C was ( 1309 ± 542 ) pg/ml in patients with advanced pancreatic cancer, which were significantly higher than that those in normal control [ (278 ±115) pg/ml, P <0.01 ]. In Cox regression, KPS score, serum CA19-9 and VEGF-C were independent factors (x2 =7.208, 6.908, 3.867, P = 0.007, 0.009, 0.049). In multivariate analysis, serum VEGF-C and KPS score were independent factors (x2 =4.873, P=0.027, x2 =5.274, P =0.022). Using serum concentration of VEGF-C at 1280 pg/ml as the cut-off point, the mean survival of patients with VEGF-C ≤1280 pg/ml was 10.0 months, and the median survival was 11.3 months, 1 year cumulative survival was 50.0% ; while they were 6.0 months, 6.3 months and 5.9% in patients with VEGF-C > 1280 pg/ml, and the difference was statistically significant (x2 = 9.400, P= 0.002). Using KPS score 70 as the cut-off point, the mean survival of patients with KPS <70was 6.0 months, and the median survival was 6.6 months, 1 year cumulative survival was 21.4% ; while they were 9.0 months, 10.1 months, 33.3% in patients with KPS score ≥70,and the difference was statistically significant (x2 =4.040, P =0.044). The difference of the median survival, 1 year cumulative survival in patients with CA19-9 ≤200 U/ml or >200 U/ml was not statistically significant (10.0 months vs. 7.8 months, 37.5% vs. 21.1% ; x2 =1910, P=0. 167). Conclusions Serum concentration of VEGF-C can used as an independent factor for predication of prognosis of patients with advanced pancreatic cancer.

Objective To investigate the effectiveness of endovascular technique in treatment of acute arterial hemorrhage of abdominal organs.Methods Totally 159 cases (145 patients) acute arterial hemorrhage of abdominal organs were enrolled.All patients underwent emergency transarterial angiography.Endovascular treatment by spring coil,gelatin sponge,or covered stent were in 40 cases.The success rate of immediate and long-term hemostasis was judged by the postoperative radiography,symptoms and the changes of vital signs and laboratory results.Results The positive signs in angiography were observed in 47 among the total 159 cases (47/159,29.56％).Seventy-seven postoperative hemorrhage cases underwent angiography and 29 cases (29/77,37.66 ％) were positive,82 non-operative hemorrhage cases underwent angiography and 18 cases (18/82,21.95％) were positive.The difference was statistical significant (P＜0.05).Endovascular treat ment were performed in 40 cases (33 by spring coils,4 by gelatin sponge and 3 by covered stent).Hemorrhages were con trolled immediately in all the 40 cases after embolization and the success rate were 100％.Conclusion Endovascular tech nique is minimal invasive,and it is an effective method for treatment of acute arterial hemorrhage in abdominal organs.

Objective To evaluate arch replacement in islet fashion combined with intraluminal-including technique.Methods From Aug.2011 to Mar.2012,19 patients with type A aortic dissection needed arch replacement were selected,whose cephalo-brachial artery and left common carotid artery were intact or left subclavicular artery alone was involved,Sixteen patients were male.Mean age was (49.6 ± 5.7) years.Urgent surgery and red surgery were completed in 13 cases,6 cases and 1 case,respectively.The arch replacement was performed in a modified islet fashion,remaining the adventitia and trimming intima,constituting a common opening with the proximate end of stented elephant trunk in side-to-side manner by using intraluminal anastomosis technique.This common opening was anastomosed with the distal end of the ascending aortic Dacron graft,completed the procedure of arch replacement.If left subclavicular artery was involved by dissection or it was displaced pathologically as to expose difficultly,it should be covered by the stented elephant trunk,and be transplanted to left common carotid artery in end-to-side manner.Finally,the entire Dacron graft was wrapped by remained adventitial coat using including technique,with a shunt connecting to right atrium.Results Mean duration of cardiac pulmonary bypass,aortic cross clamp and selective cerebral perfusion was (215 ±54),(93 ± 18) and (30 ±6) minutes,respectively.In one case with delayed sternum closure due to extensive bleeding,the mean chest tube output in first 24 hours was (926 ±322) mL,the mean duration of postoperative ICU was (78 ±21) hours; none of the patients was reopened for bleeding.Postoperative hypoxemia,transient neurologic dysfunction,and hematosepsis occurred in 3 cases,1 case and 1 case,respectively.One patient needed re-intubation attributing to pulmonary infection.There was no complication of permanent neurologic deficit or postoperative visceral mal-perfusion.All patients survived and were discharged from hospital.Severe complication was not observed at follow-up of 1 to 5 months.Conclusions The modified aortic arch replacement procedure,which adopt islet fashion combined with intraluminal inclusion technique,characterized by its feasibility,safety,reproduction and easy to control bleeding,may be considered as an alternative approach for managing arch disease.Its short-term outcomes were competitive and prospective was promising.

Objective To evaluate the long-term efficacy of antiretroviral therapy and drug resistance among human immunodeficiency virus-1(HIV-1)-infected children in Yunnan.Methods In this retrospective study,CD4+T cell counts,HIV viral loads and genetic drug resistance results were obtained from HIV-1-infected children who were treated with antiretroviral treatment between January 2004 to July 2015.Results A total of 1 078 HIV/acquired immune deficiency syndrome(AIDS)children were treated with antiviral therapy.Before treatment,the average CD4+cell number was(466.8 ± 397.2)cells/μL. The percentages of children with CD4+cell count >750 cells/μL after 1-year,3-year,5-year and 8-year treatment were 54.31%,62.87%,68.46% and 74.19%,respectively.Virological failure occurred in 150 HIV/AIDS children(13.9%),and the virological failure rate was 4.3/100 child-years.Among those 150 patients with virological failure,104 cases(69.33%)exhibited genetic resistance to antiretroviral drugs.The prevalent mutations associated with drug resistance were M 184V/I(75.0% [78/104]), K103N(43.3%[45/104]),G190A(29.8%[31/104]),Y181C(22.1%[23/104]),T215Y/F(20.2%[21/104]).Conclusions After long-term antiretroviral treatment,most of the HIV-infected children have restored the immunity and suppressed HIV viral replication successfully.HIV resistance is the main cause of virological failure.Drug resistance mutations mainly occur in nucleoside reverse transcriptase inhibitor and non-nucleoside reverse transcriptase inhibitor,and the resistance rate of proteinase inhibitor is low.Early genetic resistance testing and switch to second-line therapy will improve the treatment outcome.

Objective To investigate the distribution of chronic disease and geriatric syndrome in hospitalized elderly patients with multimorbidity by chronic disease investigation and comprehensive geriatric assessment.Methods A total of 176 patients aged ≥60 years admitted into the geriatric department in our hospital were selected,and demographic factors were collected.Their chronic diseases and geriatric syndrome were recorded.The comprehensive geriatric assessment was consummated,and the biochemical indicators for chronic disease and geriatric syndrome were detected.Results All patients suffered from two or more kinds of chronic diseases.The top three most common diseases were hypertension (112 cases),diabetes (94 cases),and respiratory diseases (73cases).The sum of chronic diseases kinds were ≥3 in 137 patients,≥4 in 78 patients,≥5 in 40 patients.The amount of chronic diseases showed an increased tendency along with ageing.Totally 175 patients had more than one geriatric syndrome,the top three most common types were frailty and prefrailty(133 cases),visual impairment (117 cases),impaired daily activities (107 cases).There were significant differences in the number of geriatric syndrome among different age groups(x2 =16.989,P ＜ 0.001).Spearman analysis showed that the number of patients with geriatric syndrome were positively correlated with age(r =0.307,P＜0.001).Conclusions The prevalence of chronic disease and geriatric syndrome is high in hospitalized elderly patients with multimorbidity,which is increased with aging.We should pay attention to the screening of chronic disease and geriatric syndrome in multimorbid elderly patients

Objective:To explore the relationship between drug resistance occurrence and the distribution pattern of polymorphic loci in individuals with human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) treated with highly active anti-retroviral therapy (HAART).Methods:HAART-failed HIV/AIDS patients who successfully amplified the gene sequences of the pol region between June 2015 and December 2021 from 16 prefecture-level administrative regions in Yunnan Province were included.The resistant sequences were classified using the human immunodeficiency virus (HIV) basic local alignment search tool (BLAST) and validated through MEGA 6.0, and the obtained sequences were submitted to the Stanford University HIV Drug Resistance Database to identify drug resistance loci. The distribution of polymorphic loci was analyzed across patients exhibiting varying degrees of drug resistance, different treatment regimens and distinct HIV-1 subtypes.Changes of the frequencies of polymorphic loci in patients with different degrees of drug resistance were analyzed using trend chi-square test. Statistical comparisons and further paired comparisons were performed using chi-square test.Results:Gene sequences were amplified from 1 453 patients, and the resistance testing results showed 954 sensitive, 224 potentially or low resistant, 189 moderately resistant, and 86 highly resistant patients. The frequencies of mutations I15V, L19I, D60E in the HIV-1 protease region (PR region) and E36A, T39D, S48T mutations in the HIV-1 reverse transcriptase region (RT region) showed a decreasing trend as the degree of HIV-1 resistance escalated ( χ2trend=19.86, 9.16, 13.66, 37.64, 18.44 and 40.86, respectively, all P<0.01). Conversely, the mutations V77I in the PR region and K122E in the RT region showed an ascending trend ( χ2trend=12.19 and 10.03, respectively, both P<0.01). Distinct treatment groups, namely zidovudine (AZT)+ lamivudine (3TC)+ lopinavir/ritonavir (LPV/r), AZT+ 3TC+ efavirenz (EFV), AZT+ 3TC+ nevirapine (NVP), and tenofovir (TDF)+ 3TC+ EFV, were examined. Statistically significant differences in the frequencies of mutations E35D, M36I, and D60E in the PR region, as well as S48T, K122E, and R211K in the RT region, were observed among these treatment groups ( χ2=22.46, 9.32, 14.46, 26.85, 18.92 and 24.26, respectively, all P<0.05). In paired comparisons, AZT+ 3TC+ LPV/r group displayed higher frequencies of E35D, M36I, and D60E mutations, the AZT+ 3TC+ EFV group showed a higher frequency of S48T mutation, the AZT+ 3TC+ NVP group showed a higher frequency of K122E mutation, and the TDF+ 3TC+ EFV group exhibited a higher frequency of R211K mutation, all with statistically significant differences (all P<0.008). The differences in the frequencies of T12S, I15V, L19I, M36I, V77I, L89M in the PR region and E53D, I135V, S162C, R211K, K277R in the RT region among circulating recombinant form (CRF)08_BC, CRF07_BC and CRF01_AE subtype group were statistically significant ( χ2=693.60, 712.51, 798.11, 434.85, 386.91, 657.78, 932.58, 409.21, 344.39, 469.44 and 260.48, respectively, all P<0.001). In paired comparisons, the frequencies of T12S, I15V, L19I, E53D, I135V, S162C and R211K in CRF08_BC subtype, the frequencies of V77I and K277R in CRF07_BC subtype, and the frequencies of M36I and L89M in CRF01_AE subtype were higher than those in the other two groups, and the differences were all statistically significant (all P<0.017). Conclusions:The polymorphic loci resulting from HIV-1 HAART failure show different distribution patterns across various degrees of drug resistance, treatment regimens and HIV-1 subtypes.These loci demonstrate both specific and shared characteristics. It is necessary to enhance the surveillance of select polymorphic loci.