Objective:To establish virus culture method and full genome sequencing method for severe acute respiratory syndrome coronavirus (2019-nCoV), and to illuminate the variation of 2019-nCoV.Methods:The pharyngeal swab specimens were inoculated into Vero-E6 cells for isolation, and the cytopathic effect were observed day by day, and the result of virus isolation were confirmed by Real-Time RT-PCR. Some isolated viruses were diluted 10 times to detect the virus titer. Full-genome of 2019-nCoV was sequenced with the whole genome capture technology and next generation sequencing technology. Phylogenetic analysis and molecular characterization for obtained 2019-nCoV sequences were undertaken.Results:Twenty-two 2019-nCoV strains were isolated with Vero-E6 cell line in the Biosafety Level-3 Laboratory (BSL-3). Eighteen full-genome sequences obtained (almost 29 000 nucleotide) were analyzed with 99.94%~100% identity and 28 nucleotides and 20 amino acids variations. Phylogenetic analysis showed that the 2019-nCoV sequences belonged to different clades, including clade S and clade L with Europe lineage L. I and Europe lineage L. II.1. Furthermore, 5 of 18 2019-nCoV harbor the D614G mutation.Conclusions:2019-nCoV were successfully isolated with Vero-E6 cells, and partially illuminated the phylogenetic characteristics, which provided a basis for subsequent drug screening and biological characteristics research. Continuous monitoring and analysis of the sequences of new cases would be vital to understand the genetic evolution and rates of substitution of the 2019-nCoV.

Objective To explore the effect of bee-needle therapy on treatment of patients with rheumatoid arthritis.Methods Seventy five patients with rheumatoid arthritis were randomly divided into bee-needle group,drug group and bee-needle plus drug group.Therapeutic effect was compared.Results After treatment and follow up,there were significant differences in degree of joint pain,the number of swollen joints,tender joint count,morning stiffness time,the number of pain joints,HAQ,ESR,RF,and CRP among three groups (P ＜ 0.05).Conclusion Bee-needle therapy can significantly relieve the clinical symptoms of patients with rheumatoid arthritis.

Objective To explore the effect of bee-needle therapy on treatment of patients with rheumatoid arthritis.Methods Seventy five patients with rheumatoid arthritis were randomly divided into bee-needle group,drug group and bee-needle plus drug group.Therapeutic effect was compared.Results After treatment and follow up,there were significant differences in degree of joint pain,the number of swollen joints,tender joint count,morning stiffness time,the number of pain joints,HAQ,ESR,RF,and CRP among three groups (P ＜ 0.05).Conclusion Bee-needle therapy can significantly relieve the clinical symptoms of patients with rheumatoid arthritis.

Objective To investigat the changes of hemodynamics and arterial blood gas during the establishment of ECMO model and provide an ideal platform for the study of the mechanism of DCD protection using ECMO, a porcine model of controlled donors of cardiac death (CDCD) with extracorporeal support was established in the paper. Methods Eight domestic crossbred pigs were anes-thetized, atracurium (1mg/kg) and heparin (150~200 U/kg) was administered and the ventilator was discontinued while the animal was under deep anesthesia to kill the animal.Meanwhile, blood was drained from the left jugular vein catheter into the ECMO circuit and re-turned into the left femoral artery to establish the porcine model of CDCD with extracorporeal support . The average blood press and arterial blood gas were ministered at normal, cardiac arrest and ECMO supporting 1 h, 3 h and 5 h. Results The model of CDCD with extra-corporeal support was established successfully.Two animals restored the beating of the heart and the high doses of potassium chloride were given untill cardiac arrest during extracorporeal support.Blood gases throughout the apneic time showed a severe hypoxemia and acidosis, pH and PaO2 reduced to 7.09 ±0.24 and 5.3 ±0.93mmHg respective-ly, which can be improved during extracorporeal support.MAP and PaO2 were maintained over 300mmHg and 65mmHg respectively. Conclusion A new porcine model of controlled donors of cardiacdeath with extracorporeal support has been successfully established and showed to be feasible for related pathophysiological research work.

Objective To evaluate the effect of extracorporeal membranous oxygenation (ECMO) on the small bowel preservation in a porcine model of controlled donors after cardiac death (CDCD).Method The domestic crossbred donor pigs were anesthetized and ventilated with 100％ oxyger.An intravenous cannula was placed through iliac arteries and jugular vein,and connected to ECMO system.The ECMO was performed to infuse abdominal organs when cardiac death was declared.Pathology,electron microscope,energy metabolism and cell apoptosis level of intestinal mucosa were evaluated before operation,in cardiac death and at the end of extracorporeal support,respectively.Result The normothermic extracorporeal support could quickly restore graft blood supply and oxygenation.One-h extracorporeal support could improve the energy status in intestine of donation after cardiac death (DCD).The histologic damage and apoptosis of 1-h extracorporeal support had no significant difference with those before operation and in cardiac death (P＞0.05).With the extension of extracorporeal support,the intestinal mucosa damage degree was gradually increased,and the content of adenosine triphosphate in intestinal mucosa reduced gradually.Conclusion The normothermic extracorporeal support for 1 h in DCD is beneficial for improving the viability of small bowel.However,the integrity of intestinal mucosa is destroyed gradually as extracorporeal supporting time over,which may be initiated by the activation of intestinal epithelial apoptosis.

OBJECTIVETo investigate the correlation between Shigella flexneri multi-drug resistance and drug resistance gene cassette of integrons.

METHODAll 79 strains of Shigella flexneri were isolated from the feces of children ranged in age from 6 months to 14 years in some hospitals of Jinan, between May 2009 and April 2012.The resistance was detected by Kirby Bauer agar diffusion method, 1, 2 and 3 integron gene was amplified by PCR, the variable region of positive strains treated with enzyme digestion and determined by Series Analysis.

RESULTAmong 79 Shigella flexneri strains, the resistance rate was 91% (72/79) to ampicillin, chloramphenicol, tetracycline, streptomycin, 70% (55/79) to sulfamethoxazole/trimethoprim, 30% (24/79), 23% (18/79), 33% (26/79) and 32% (25/79) to cefotaxime, ceftazidime, ciprofloxacin and levofloxacin.All 79 strains were susceptible to cefoxitin, imipenem, cefoperazone/sulbactam. The common drug resistance pattern is ampicillin tetracycline-chloramphenicol-streptomycin, accounted for 91% (72/79); 91% (72/79) strains carried integrons of class 1, 86% (68/79) strains carried integrons of class 2, No intI3 was detected. The resistance to ampicillin, streptomycin, tetracycline, chloramphenicol of atypical class 1 integron positive strains was significantly higher than the negative strains (χ² = 35.96, P<0.01). The sequencing results:dfrV was detected in class 1 integron variable regions of 9 strains, dfrA17-aadA5 in 2 strains, blaOXA-30-aadA1 in 70 strains, 2 strains were not detected resistance gene cassette, all resistance gene cassettes were dfrA1-sat1-aadA1 in class 2 integron variable regions.

CONCLUSIONThe muti-drug resistance of Shigella flexneri in Jinan was closely associated with integrons.

Adolescent ; Anti-Bacterial Agents ; pharmacology ; Child ; Child, Preschool ; Drug Resistance, Multiple, Bacterial ; genetics ; Dysentery, Bacillary ; microbiology ; Feces ; microbiology ; Humans ; Infant ; Integrons ; Polymerase Chain Reaction ; Shigella flexneri ; drug effects ; genetics

OBJECTIVETo analyze the impact of atorvastatin on blood lipids and arterial media thickness (IMT) in new-onset type 2 diabetes patients.

METHODS333 patients, 30-70 years old and diagnosed within one year as type 2 diabetes, were selected from the Chinese Diabetes Complication Prevention Study (CDCPS) to take part in this study. Changes of blood lipids and IMT of carotid, femoral and iliac artery pre and post the administration of atorvastatin were tested and followed for 24 months.

RESULTSTotal cholesterol, triglycerides and low-density lipoprotein decreased significantly (P = 0.000) and maintained at a low level. The carotid artery IMT decreased significantly (P = 0.022) at the end of this study, but the femoral and iliac artery IMT did not show any obvious change. There were no serious adverse events noticed, during the study period.

CONCLUSIONLong-term use of atorvastatin seemed to be safe and effective in reducing blood lipids in patients with type 2 diabetes thus could delay the development of atherosclerosis.

Adult ; Aged ; Atorvastatin Calcium ; Diabetes Mellitus, Type 2 ; drug therapy ; Female ; Follow-Up Studies ; Heptanoic Acids ; therapeutic use ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Pyrroles ; therapeutic use ; Tunica Media ; pathology

Objective To investigate the prevalence and characteristics of thyroid nodule and its relationship with thyroid function in a group of middle-aged to elderly individuals.Methods A cross-sectional,stratified,and cluster random sampling study was performed in 1 721 subjects (average age 71.4 years,from 40 to 108 years).All subjects underwent ultrasound thyroid scan,and the prevalence of thyroid nodule and its size,quantity,property,and distribution were age-stratified and analyzed.The relationship between nodule existence and thyroid function was assessed in subjects who randomly accepted TSH,FT4,and FT3 tests.Results The prevalence of thyroid nodule reached to 76.6％.The incidence of nodule increased with age (x2 =92.08,P＜0.01),and was more common in female (87.4％ vs 75.5％,P =0.001).In patients with thyroid nodule,the prevalence of solitary nodule was 25％,compared with 75％ of multiple nodules,which increased with age (x2 =78.55,P＜0.01).The prevalence of small nodule reached to 68.2％,compared with 31.8％ of large nodules,which also increased with age (x2 =36.82,P＜0.01).As to the property of nodule,solid nodule was the most common form (72.3 ％),cyst-solid nodule accounted for 26.6％,and detection rate of cyst nodule was only 2.1％,and there was no significant difference among the different age groups (x2 =14.22,P=0.076).884 among the total subjects underwent thyroid function test,multiple logistic regression analysis demonstrated that the reduction of TSH was the only independent predicting factor of thyroid nodule (P =0.005).Conclusions The prevalence of thyroid nodules was considerably high according to the examing of ultrasonographic image,especially in the elderly.The incidence of thyroid nodule,especially multiple nodules or larger nodule,increased with age and solid nodule was the most common finding.

Objective To analyze the impact of atorvastatin on blood lipids and arterial media thickness(IMT)in new-onset type 2 diabetes patients. Methods 333 patients,30-70 years old and diagnosed within one year as type 2 diabetes,were selected from the Chinese Diabetes Complication Prevention Study(CDCPS)to take part in this study. Changes of blood lipids and IMT of carotid, femoral and iliac artery pre and post the administration of atorvastatin were tested and followed for 24 months. Results Total cholesterol,triglycerides and low-density lipoprotein decreased significantly (P=0.000)and maintained at a low level. The carotid artery IMT decreased significantly(P=0.022) at the end of this study,but the femoral and iliac artery IMT did not show any obvious change. There were no serious adverse events noticed,during the study period. Conclusion Long-term use of atorvastatin seemed to be safe and effective in reducing blood lipids in patients with type 2 diabetes thus could delay the development of atherosclerosis.

Objective To investigate the effects of metformin on blood glucose control and body weight management in over-weight patients with newly diagnosed type 2 diabetes mellitus (T2DM).Methods A retrospective study was performed in T2DM patients who were treated with metaformin (male 38,female 23,mean age (50.8 ± 7.9) years old).Based on body mass index (BMI),all the participants were assigned to obese,over-weight and normal weight group.Analysis of variance and multivariate analysis were used to confirm the effects of metformin on blood glucose and body weight.Results Fasting blood glucose (FBG),HbA1 c and BMI were significantly reduced over 2 years.In comparison with baseline,BMI of the obesity and overweight group was significantly decreased during follow up (P ＜ 0.05),while BMI of the normal weight group increased by 0.7 kg/m2.At baseline,FBS and HbAlc of the obese group were lower than those of the normal weight group (FBS (7.3 ± 0.8) vs.(11.8 ± 4.8) mmol/L,P ＜ 0.05 ; HbA1 c (6.8 ± 1.2) ％ vs.(8.5 ±3.1)％,P ＜0.05),whereas insulin level was higher than that of the normal weight group ((2.20 ± 0.36) vs.(1.87 ± 0.29) U,P ＜ 0.05).At 2 years,HbA1 c was reduced on average by 2.5％,1.4％ and 1.9％ in the obesity,over-weight and normal weight group,respectively.Conclusions The present study demonstrates the beneficial effect of metformin on newly diagnosed diabetics,especially obese patients.