Objective To investigate the resistant status and to type the phenotype and plasmid profiles of prevalent gonococcal strains. Methods A total of 98 gonococcal strains isolated from 1998 to 1999 in Zhanjiang was identified systemically. The susceptibilities of 98 strains of gonococci to penicillin,tetracycline and ciprofloxacin were detected by agar dilution method. The β － lactamase production was tested by iodometric method. The plasmids were extracted by alkaline lysis technique. Results The prevalence of gonococcal strains resistant to penicillin, tetracycline and ciprofloxacin were 22.44％ ,45.94％ and 55.12％ ,respectively. There were 6 positive strains for β － lactamase, accounting for 6.12％ (6/98). The most common phenotypes were susceptiblePT、 CMRNGT、 CMRNGPT and TRNG, accounting for 43.86％ ,23.46％ ,10.20％ and 10.20％ ,respectively. The detection rate of plasmids were 91.84％ .Among them, the percentages of 42.5 kb,39.5 kb,7.4 kb and 4.2 kb plasmids accounted for 11.22％ ,41.82％ ,59.16％ and 67.32％ ,respectively.There were 12 types of plasmid profiles(PP), the predominant types were 7.4kb＋ 4.2kb and 39.5kb＋ 7.4kb＋ 4.2kb, accounting for 21.42％ and 17.34％ ,respectively. Conclusion The gonococcal strains resistant to ciprofloxacin are very high in Zhanjiang. The phenotype and plasmid profiles of the strains have characteristic distribution.

Objective To prepared inclusion complex of β-cyclodextrin (β-CD) and its derivatives (methyl -β-cyclode-xtrin, 2,6-dimethyl -β-cyclodextrin, hydroxypropyl -β-cyclodextrin ) with entrcavir, and explore effect of different inclusion complex on the solubility and dissolution of entecavir.Methods Regression equation was established,prepared the inclusion complexes by a magnetic stiring method,grinding method and ultrasonic method.While inclusion rate was an index,the effects of reaction process was inquired by mole ratio, inclusion temperature, inclusion time and stirring rate and the most suitable method and technology of inclusion was optimized.Determinated the solubility with HPLC method and investigated in vitro dissolution with a small cup method.Results The regression equation was A=1.026 ×106C+4248.8, R2 =0.9999 (n=6), in the range of 0.497~4.97 g/mL, linear relationship was good.The best inclusion method was magnetic stirring, the best process conditions were: mole ratio was 1 ︰1, stirring speed was 300 r/min, inclusion time was 4 hours, inclusion temperature was 50 ℃.The four inclusion complexes all were white loose powder materials.The solubility of ETV,β-CD-ETV,HP-β-CD-ETV,RM-β-CD-ETV,2,6-DM-β-CD-ETV were 2.51, 13.4, 18.9, 37.6 and 89.5 g/L.The dissolution rates was ETV<β-CD-ETV

In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.

Objective To investigate the association between the apolipoprotein E (apoE) gene polymorphism and the dose for warfarin individual maintenance. Methods The genotypes of 249 patients with warfarin treatment in maintenance doses were determined by PCR/DHPLC assay. The doses for warfarin maintenance were compared among patients with different genotypes. Results In the total of 249 patients, the frequencies of 2/ε2, ε2/ε3, ε2/ε4, ε3/ε3, ε3/ε4, ε4/ε4 genotype were 1.20%, 15.66%, 1.80%, 72.29%, 9.24%, 0.80%, respectively; the allele frequencies of ε2, ε3, ε4 were 9.44%, 84.74%, 5.82%, respectively. The warfarin dose of group ε2 (ε2/ε2, ε2/ε3) was (3.24±1.36) mg/d, slightly higher than that of group ε3 (ε3/ε3, 2.91±1.14 mg/d) or group ε4 [ε4/ε4, ε3/ε4, (2.98±1.05) mg/d], but the difference of the warfarin doses among the 3 groups did not reach statistical significance (F=1.848,P＞0.05). Conclusion ApoE polymorphism may be not a major genetic factor that influences the individual dose for warfarin maintenance.

Objective To investigate the value of femoral neck fracture(FNF) typing by digital subtraction angiography(DSA) in prognostic prediction of the fracture healing. Methods From June 2005 to June 2008,45 patients were admitted to our hospital for treatment of single femoral neck fracture.They were 26 males and 19 females,aged from 26 to 60 years(average,45.4 years).Their fractures were all typed according to DSA results.Type Ⅰ ( 15 patients) were DSA pictures which showed 3 to 6 supporting band vascular images crossing the fracture line.Type Ⅱ(14 patients) were DSA pictures which showed one to 2 supporting band vascular images crossing the fracture line.Type Ⅲ(16 patients) were DSA pictures which showed no supporting band vascular images crossing the fracture line.All cases underwent close reduction and internal fixation with cannulated screws under C-arm fluoroscopy.The prognoses of all the patients were evaluated according to their symptoms and imaging examinations in terms of avascular necrosis of femoral head (ANFH) and fracture healing. Results All the patients obtained an average follow-up of 36.7 months (from 24 to 60 months).In patients of DSA type Ⅰ,the ANFH rate and nonunion rate were respectively 0 and 13.3％ (2/15).In patients of DSA type Ⅱ,the ANFH rate and nonunion rate were respectively 7.1％ (1/14) and7.1％(1/14).In patients of DSA type Ⅲ,the ANFH rate and nonunion rate were respectively 100％ (16/16) and 0.The ANFH rate following FNF treatment was negatively related to age and the vascular number showed by DSA.The nonunion ratewas positively related to age but not significantly associated with the vascular number showed by DSA. Conclusion DSA has a significant value in predicting the prognosis for patients with femoral neck fracture.

Objective To study the characteristics of neutralization antibody in mice induced by DNA vaccines of hemagglutinin(HA) of novel H1N1 influenza A virus(2009H1N1).Methods HA encoding plasmids of 2009H1N1 or 1918H1N1(2009HA or 1918HA)were constructed.25 μg or 200 μg dosage of 2009HA plasmids were used to immunize the mice,the total antibody of anti-20O9HA or cross-reactive antibody were assayed by ELISA using 2009HA or 1918HA protein as capture antigen,and the neutralizing antibody were assayed by two kinds of virus pseudo - particles(pp) of 2009H1N1 and 1918H1N1 .Results During of 4 to 16 weeks after boost immunization,in two groups of mice immunized with 25 μg or 200 μg dosage 2009HA plasmids,both total antibody of anti-2009HA and neutralizing antibody to 2009H1Nlpp reached the similar level(P ＞0.05),and there were cross-reactive antibody to 1918HA protein in two groups of mice serum,with similar titers of cross-neutralizing activity to 1918H1N1 pp(P ＞0.05),Conclusion A low dosage DNA vaccine encoding HA of 2009 H1 N1 virus is able to induce persistent and high level of neutralizing antibody,and may be potential valuable vaccine against the new emerging influenza virus.

Objective: To create and validate a scoring system for predicting clinical prognosis in patients with cardiac resynchronization therapy (CRT). Methods: A cohort of 367 consecutive patients received CRT in our hospital from 2010-01 to 2015-12 were enrolled. The endpoint follow-up events were all-cause death including heart transplantation and heart failure re-admission. The patients were randomly categorized into 2 groups: Modeling group, to develop HEAL scoring system,n=300 and Veriifcation group, to validate HEAL model,n=67. HEAL system was established by Cox proportional hazards regression model, discrimination between HEAL and EARRN scoring systems was evaluated by AUC of ROC, HEAL calibration was assessed by Hosmer-Lemeshow test and clinical endpoint evaluation by 2 scoring systems were compared by Kaplan-Meier method. Results: Modeling group analysis indicated that hs-CRP (HR=1.137, 95% CI 1.072-1.205,P<0.001), big endothelin-1 (HR=1.934, 95% CI 1.066-3.507,P=0.03), left atrial diameter (HR=1.045, 95% CI 1.007-1.084,P=0.02) and NYHA IV (HR=2.583, 95% CI 1.331-5.013,P=0.005) were the independent risk factors of adverse prognosis in CRT patients. Based on β partial regression coefifcient, HEAL scoring system was established to classify the patient's risk levels: low risk<4, moderate risk 4-10 and high risk>10. AUC for risk classification in Modeling group and Verification group were 0.719(95% CI 0.629-0.809) and 0.708 (95% CI 0.539-0.878), HEAL can well distinguish clinical prognosis in patients at different risk levels (log-rank test showed in Modeling groupP<0.001 and in Veriifcation groupP=0.002); Hosmer-Lemeshow test presented good calibration,P=0.952. All 367 patients were respectively evaluated by HEAL and EARRN scoring systems, HEAL had the better discrimination than EARRN as AUC 0.763 (95% CI 0.692-0.833) vs AUC 0.602 (95% CI 0.517-0.687). Conclusion: HEAL scoring system can effectively predict adverse prognosis in CRT patients, it had the better discrimination than EARRN system and was valuable to distinguish high risk patients in clinical practice.

AIM:To investigate the effect of perifosine, an inhibitor of protein kinase B ( PKB/Akt) , on the cell cycle, apoptosis and autophagy in human brain glioma U251 cells, and to determine the relationship between perifos-ine-induced autophagy and apoptosis of glioma.METHODS:The cell growth inhibition was determined by MTT assay. The cell cycle distribution of U251 cells was examined by flow cytometry.The cell apoptosis was analyzed by Annexin V-FITC apoptosis detection kit.The protein expression of P21, P27, cyclin B1, caspase-9 and PARP was examined by Wes-tern blot analysis.The distribution and expression of LC3-Ⅱ, an autophagy marker, was observed to determine the effect of perifosine-induced autophagy.RESULTS:Perifosine inhibited the cell viability in a dose-dependent manner.In perifos-ine-treated U251 cells, the cell cycle was arrested in G2 phase and the expression of cyclin B1 was inhibited.Perifosine in-duced apoptosis of U251 cells through activation of caspase-9 cleavage, PARP cleavage and survivin inhibition.In addi-tion, suppression of autophagy by chloroquin, an inhibitor of autophagy, increased the number of apoptotic cells.CON-CLUSION:Perifosine inhibits cell proliferation and triggers apoptosis and autophagy in human U251 cells.Blocking auto-phagy magnifies perifosine-induced glioma cell apoptosis.

Objective:To illustrate a relatively complete knowledge system (e.g., research outputs, current hotspots, and future trends) in the sepsis field and to help scholars grasp the scientific research direction or clinical focus of treatment.Methods:The relevant literatures of sepsis during the time from 1985 to 2019 in Web of Science database were collected. Sepsis-related research contents were generated using softwares (CiteSpace 5.6.R2 and VOSviewer 1.6.13), which using data mining, information processing and knowledge map methods, to analyze the historical evolution and predict the development trend.Results:A total of 8 189 papers on sepsis were published. The volume of publications were increasing yearly from 1985 to 2019, and reached the top list of 1 276 in 2019. For research contents of sepsis, it has formed the basic characteristics of sepsis which focusing on epidemiological studies and animal experiments. Through cluster analysis, the researches mainly focused on six aspects: septic rat, necrotizingenterocolitis, sepsis-associated encephalopathy, acute kidney injury (AKI), gut-derived sepsis, and inflammatory mediator. And it presented the literature characteristics that related to the injury or dysfunction of intestines, brain, liver, kidney or other organs, but the heart and lung researches were more marginal. Additionally, based on the top key words with the strongest citation bursts, it reflected that the development trend of the continuous attention hotspots with "endotoxin" or "endotoxin shock", the significant attention hotspots with "inflammation", "immunity" and "multiple organ dysfunction syndrome" (MODS), and the novel burst attention hotspots with sepsis management including "diagnosis" and "chemotherapy".Conclusions:Through the hotspots and trends visualization of sepsis, the current researches are prefer to animal experiments, epidemiology, or other basic scientific aspects. Meanwhile, the researches are mostly focusing on inflammatory reaction, immune function or organ dysfunctions. Integrating the knowledge maps of hotspots and trends, based on researches of epidemiology, diagnosis, risk factors, pathogenesis, or treatment, we predict that the future scientific topics will concentrating on childhood sepsis, organ injury mechanism or intervention relating to MODS, and integrated management of sepsis by combining traditional Chinese medicine and Western medicine.

Objective To establish an external quality assessment ( EQA) system of genitourinary tract secretions routine testing in Guizhou Province and improve the overall testing level.Methods From 2009 to 2011, more than 50 clinical laboratories in different grade hospitals from Guizhou Province were enrolled as participating units every year.EQA was carried out twice a year.Each time, five slides of high quality Wright′s or Gram stain smear of the genitourinary tract secretions or photographs obtained from these smears were selected to send to the participating laboratories for testing, and the feedback results from each laboratory were analyzed.The qualification was judged by the coincidence rate equal to or more than 80%. The average coincidence rates of each time and each year were statistically analyzed by Chi-squared test. Results From 2009 to 2011, the number of EQA participating units increased from 55 to 96, with an average return rate of >80%.Coincidence rates <80%of the 6 EQA results in the 3 years were as follow:four times for coccobacteria (73.7%,77.8%,61.1%,77.1%), twice for bacillus (75.6%,79.3%) and coccobacillus (64.3%,52.1%), once for infusorian (79.7%), epithelial cells (76.1%), neutropenia (75.7%) and cleanliness (71.3%).There were six batches of 30 quality assessment controls (accounting for 20.0%) in the six EQAs had the coincidence rate of <80%.Eleven items of 30 quality assessment controls with 1 to 15 batches were unqualified ( average coincidence rate of<80%) respectively.The item with the highest total average coincidence rate was suspected gonococcus (94.2%), and two items with the lowest total average coincidence rates were coccus and coccobacillus ( 77.0%, 75.2%, respectively ) . Conclusions This EQA program carried out within a certain range of clinical laboratories achieved good results:participating units increased significantly;the total score of all the items showed an obviously upward trend;the quality awareness of clinical lab technicians has enhanced to a certain extent.In this study, EQA system of genitourinary tract secretion routine testing were preliminarily established in Guizhou province, which provided a reference model of internal quality control ( IQC ) and EQA for clinical laboratories and higher authorities, and will be bound to have a positive impact on improvement of the overall level of genitourinary tract secretion routine testing.