Objective To investigate the characteristics of strains of AmpC β-lactamase(AmpC enzyme)production in Dathogenic bacteria in blood stream and clinical presentations of the cases, and study the related ampC and ampD genes.Methods One hundred and eighty-one strains of gram negative bacillus in blood stream were collected,Cefoxitin screening test and three-dimensional test were performed for screening of strains of AmpC enzyme,production and those of AmpC enzyme hyperproduction retrospective analysis was condected in the strains with positive results.ampC and ampD gene PCR ampliftcation, sequencing and sequence analysis of positive strains were performed, and gene homology of ampC positive strains was analysed bv Rep-PCR. Results Among 181 strains in blood stream,strains of AmpC enzyme production were detected in 39 isolates by Cefoxitin screening test,with the detection rate of 21.5%(39/181).The detection rate of strains of AmpC enzyme hyperproduction by three-dimensional test was 43.6%(17/39).PCR revealed that the positive rates for ampC and ampD genes were 41%(16/39)and 56.4%(22/39),respectively.The ampC gene sequencing of 16 positive strains indicated that the homology was 98%to 100%by comparison with the GenBank,while the ampD gene sequencing of 2 strains of Enterobacter cloacae demonstrated that the suspected gene mutations existed in the carboxy-terminal of ampD gene. Conclusion The prevalence of drug-resistant pathogenic bacteria in blood stream in this study is due to nosocomial infection.The mutation of ampC gene is rare in the pathogenic bacteria in blood stream with production of AmpC enzyme,while the rate of gene mutation in Enterobacter cloacae is higher, and the deletion and amino acid substitutions in the carboxy-terminal of ampD is highly relevant to the depressed expression of AmpC enzyme.

ObjectiveTo investigate the inhibitory effect of dacarbazine and an oncolytic adenovirus carrying interleukin-24 (IL-24) on transplanted melanoma in nude mice.MethodsNude mice were inoculated with human A375 melanoma cells to establish a model of malignant melanoma.Then,the mice were divided into 4 groups to be treated with an oncolytic adenovirus carrying interleukin-24 (ZD55-IL-24),dacarbazine,the combination of ZD55-IL-24 and dacarbazine,and phosphate buffer(PBS),respectively,for 3 days.Seven days after the end of the treatment,some mice were sacrificed followed by the determination of IL-24 and E1A protein levels in tumor tissue by Western blot.The tumor volume was measured on a daily basis for 30 days.ResultsIL-24 and E1A were highly expressed in melanoma cell-bearing nude mice treated with ZD55-IL-24 and dacarbazine.At 30 days after the inoculation,the average volume of transplanted melanoma was (2346.5 ± 576.0) mm3 in the combination group,significantly different from that in the ZD55-IL-24 group((4141.6 ± 1348.2) mm3,P ＜ 0.05),dacarbazine group((5230.1 ± 922.8) mm3,P ＜ 0.05),and the control group ((7135.1 ± 1002.3) mm3,P ＜ 0.05).ConclusionThe ZD55-IL-24 in combination with dacarbazine exhibits a remarkably inhibitory effect on the proliferation of melanoma transplanted into nude mice.

Objective: To observe the clinical effect of tuina plus medications on infantile diarrhea induced by rotavirus infection. Methods: After 55 cases of confirmed sick infants were divided into two groups by the order of their visits, 30 cases in the medication group were treated by intravenous infusion of Ribavirin and oral administration of Smecta; 25 cases in the tuina plus medication group were treated by the manual techniques of tonifying Pijing(脾经) and clarifying Dachangjing (大肠经), rubbing the abdomen and kneading the navel clockwise, pushing Shangqijiegu (上七节骨), kneading Guiwei (龟尾), and pinching the skin of the spine, in combination with same medications used as in the medication group. Results: The total effective rate was 96% in the tuina plus medication group, P < 0.01, in comparison with the medication group. Conclusion: tuina has a good therapeutic effect in the treatment of infantile diarrhea induced by rotavirus infection and is importantly significant for shortening the course, enhancing the therapeutic effect and lowering down the medical cost.

The current treatments of metastatic malignant melanoma include chemotherapy,targeted therapy,immune therapy and radiation therapy,but the treatment outcome is far from optimism.In order to im-prove the treatment efficiency,it is urgent to improve early diagnosis,and develop more effective treatment drugs and delivery systems.The application of nanotechnology in the diagnosis and therapy of melanoma can re-duce the resistance to the drugs,increase efficacy and reduce side effects.

Objective To investigate the relationship between anti-HCV antibody level and hepatitis C virus genotype in the patients.Methods Total of 603 anti-HCV positive serum samples were collected during 2013 to 2014 by retrospective research method.HCV RNA were detected in anti-HCV positive samples by repeat test and the genotype were detected in HCV RNA positive samples.The distribution of anti-HCV level in different hepatitis C genotype patients was analyzed and the body's response to viral antibodies and viral genotype correlation with anti-HCV concentration interquartile range was explored.Rates among genotype groups were compared using chi-square test.Results Totally 412 of 603 (68.33％) samples were anti-HCV positive by double reagent screening.174(42.3％) samples were detected as HCV RNA positive.The distributions of different anti-HCV level in different genotype patients were 1a(n =8) 1/8,1/8,4/8,2/8;1b(n =112)25.9％ (29/112),17.0％ (19/112),25.9％ (29/112),31.3％ (35/112);2a(n =14)3/14,4/14,5/14,2/14;3a(n =11)3/11,6/11,2/11,0/11;3b(n =16)4/16,11/16,1/16,0/16;6a(n =8)2/8,2/8,1/8,3/8 with anti-HCV concentration interquartile range respectively.The anti-HCV concentration distribution was different in patients with different HCV genotypes.The anti-HCV concentration distribution in patients of 1 b,2a and 6a genotypes were evently,while anti-HCV level was relatively high in 1a (13.65) and relatively low in 3b (8.77).There were differences in different genotypes of antibody concentrations (x2 =35.2,P ＜ 0.05).Conclusions There was correlation between anti-HCV level and HCV genotype.Because there were fewer cases in some genotypes,it was necessary to investigate more samples to corfirm the above results.

Objective To understand the drug-resistance of MRSA patients in neurosurgery intensive care unit,raise the prevention of MRSA and provide doctors the basis for controlling it. Methods The 5 year(20012005) MRSA patients were tested by Kirby-Bauer in neurosurgery intensive care unit of a third-tier general hospital in Shanghai. Statistic and analysis the drug-resistance of the patients. Results The rates of 121 MRSA drug-resistance to penicillin G,erythromycin, ciprofloxacin, amikacin and the cephalosporins are 92.3 % to 100 %, totally senaitire to teicoplanin and vancomycin and lower drug-resistance to rifampin,netilmicin and fosfomycin, but it rapidly raised from 10.0 % (2001 ) to 95.2 % (2005) to sulfamethoxazole. Conclusion It is time to take care of the drug-resistance of MRSA. Prevention and use antibiotics properly are the important ways to decrease the hospital infection and to improve the quality of recovered.

Objective To investigate the effect and mechanism of valproic acid in preventing astrocyte proliferation around the central canal of rats following spinal cord injury.Methods Forty-five Wister rats were divided into normal control group (n =5),injury group (n =20) and treatment group (n =20) according to random number table.Animal models of acute spinal cord injury were produced at T10 using Allen' s method by dropping a 10 g weight from a 15 mm height.Rats in treatment group received intraperitoneal injection of valproic acid (300 mg · kg-1 · d-1 in two divided doses) at 30 minutes postinjury.Instead,rats in injury group were injected with an equal volume of saline in the same way.Hindlimb function was evaluated using BBB scoring system at 1,3,7,and 14 days postinjury.Astrocytes proliferation around central canal and expression of glial fibrous acid protein (GFAP) were examined.Results In normal control group,few astrocytes around spinal central canal and a low expression of GFAP were detected.In injury group,astrocytes began to increase at 24 hours postinjury; fluorescence intensity for GFAP was 24.6 ± 3.6 at 24 hours,reached a peak of 69.2 ± 6.4 at 3 days,maintained a high level of 56.7 ± 5.6 at 7 days,and reduced to 35.4 ± 4.3 at 14 days,a level that remained higher than that in normal control group (11.2 ± 1.6).Whereas in treatment group at 3 and 7 days,astrocyte proliferation around spinal central canal was lower than that in injury group; GFAP expressions (47.8 ± 5.3 and 42.2 ± 6.7) were lower than those in injury group (F =177.6,P ＜ 0.05).At 3,7,and 14 days,BBB scores in treatment group (7.80 ± 0.83,12.00 ± 1.58,and 16.60 ± 1.12 respectively) were significantly higher than those in injury group (4.60 ± 0.54,6.65 ± 0.67,and 9.40 ± 1.14 respectively) (F =1 113.6,P ＜ 0.05).Conclusion After spinal cord injury,valproic acid reduces astrocyte proliferation around central canal via inhibiting GFAP expression to promote functional recovery.

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.

Objective To investigate the effect of Lactobacillus plantarum(LP)on intestinal flora and bacterial translocation in mice with spontaneous inflammatory bowel disease(IBD). Methods Interleukin 10 knockout mice(IL-10~(-/-))were used as models of IBD.Eight-week old female mice were randomized to control group, IL-10~(-/-)group and IL-10~(-/-)+LP group.IL-10~(-/-)+LP group received 0.5 mL LP(1.0×10~9CFU/mL)per day for 4 weeks,and the other groups received 0.5 mL Ringer buffer.Intestinal flora including Bifidobacteria,Lactobacilli,Enterobacteriaceae and Clostridium perfringens in the feces and bacterial translocation in mesenteric lymph nodes and spleens were detected. Results The contents of Bifidobacteria and Lactobacilli significantly decreased in the intestine of IL-10~(-/-)mice,while those of Enterobacteriaceae and Clostridium perfringens significantly increased,and the bacterial translocation significantly increased.Four weeks after LP treatment, the disturbed intestinal flora was restored, and the bacterial translocation decreased. Conclusion LP administration can modulate the imbalance of intestinal flora and decrease the bacterial translocation,thus enhance intestinal barrier function in mice with IBD.

Objective To investigate the production of carbapenemase and 16S rRNA methylase in five isolates of pan-drugs resistant E.cloacae recovered in Ruijin hospital.Methods MICs of the five isolates to 10 antibiotics were determined by E test.Six kinds of 16S rRNA methylase genes and a series of β- lactamase genes were amplified by PCR.Shotgun cloning was performed to detect carbapenem resistance determinant.The conjugal transfer of carbapenemase gene and 16S rRNA methylase gene was performed in broth culture with E.coli J53 as the recipient.Pulsed-field gel electrophoresis (PFGE) was carried out to analyse the genotyping.IEF was performed to detect β-1actamases.Southern blot was performed to determine the location of carbapenem resistance determinant.Results The MICs of 10 antibiotics were ＞32 mg/L.Four β-1actamases with pIs of 5.4 ( TEM-1 ),6.7 ( KPC-2 ),8.2 ( SHV-12 ),8.4 (CTX-M-14) were determined.The insertion sequence in the recombinant plasmid was blaKPC-2 flanked by a transposon.blaKPC-2 was located on a large non-conjugative plasmid whereas armA was located on an other conjugative plasmid.PFGE patterns of 5 isolates were identical.Conclusion KPC-2 was responsible for carbapenem resistance in pandrugs resistant Enterobacter cloacae.There was no relationship between blaKPC-2 and armA.Although pandrug resistant Enterobacteriaceae remain rare,the emergence of this group of organism merits monitoring.