OBJECTIVE:To analyze the distribution and drug resistance of pyogenic encephalitis in our hospital,and to pro-vide reference for rational use of antibiotics. METHODS:The cerebrospinal fluid pathogen of 4 255 patients with pyogenic encepha-litis in our hospital during Jan. 1st,2011-Dec. 31st,2014 were cultured and identified,and drug sensitivity test was conducted. RE-SULTS:A total of 834 pathogens were isolated with positive detection rate of 19.6%,including 576 strains of gram-positive bacte-ria,accounting for 69.1%;255 strains of gram-negative bacteria,accounting for 30.6%;3 strains of fungi,accounting for 0.4%. Top 3 gram-positive bacteria were Coagulase-negative staphylococci(436 strains),Staphylococcus aureus(56 strains)and Entero-coccus (29 strains). Top 3 gram-negative bacteria were Klebsiella pneumoniae (46 strains),Acinetobacter baumannii (38 strains) and Escherichia coli(31 strains). Top 3 departments were neurosurgery department(506 strains),ICU(169 strains)and severe re-spiratory disease department (64 strains). Results of drug sensitivity test showed that no drug-resistant Staphylococcus and Entero-coccus strains to vancomycin,teicoplanin and miuocycline was found;nonfermenting Gram-negative bacilli showed low resistant to minocycline. CONCLUSIONS:Gram-positive bacteria dominates the detection rate of cerebrospinal fluid of pyogenic encephalitis patients;drug resistance of various pathogens is serious and clinicians should choose antibiotics based on drug sensitivity.

China ; Humans ; Orphan Drug Production

Objective:To construct the lentiviral-vector encoding human interleukin-10 protein(LV-hIL-10) and to observe the effect of LV-hIL-10 on controlling neuropathic pain via intrathecal administration in CCI rats. Methods:hIL-10 gene fragment was isolated and amplified from pCYIL-10 plasmid by PCR, and was cloned into pWPXL. The recombinant plasmid pWPXL-IL-10,envelope plasmid pMD2.G and packaging plasmid psPAX2 were cotransfected into 293T cells, LV-hIL-10 is prepared by concentrating the collected supernatant .At the same time, the empty plasmid pWPXL-GFP,pMD2.G and psPAX2 were cotransfected into 293T cells, LV-GFP is prepared for contrast.135 sheer breed pathogen-free adult male Sprague-Dawley rats divided into 9 arrays at random: CCI models 4 arrays (C0,C1,C2,C3), sham operatived rats 4 arrays (S0,S1,S2,S3) and a normal contrast array (N), each respectively intrathecal injection LV-hIL-10 (C1,S1)、LV-GFP (C2, S2),isotonic Nachloride (C3,S3) and control (no implanted catheters and no administration, C0,S0), the pain threshold of each array and the expression of mRNA and protein of IL-10 in spinal cord,pallium and hippocampus on different time were observed after intrathecal administration LV-hIL-10 in successful CCI model rats . Results:The hIL-10 gene fragment was obtained from pCYIL-10 plasmid, pWPXL-hIL-10 was recombinated successfully. the cloned gene segment was validated by DNA sequencing .High titer(2?1010)and highly purified LV-hIL-10 particles were obtained by three plasmids were cotransfected into 293T cells. The mechanical allodynia and thermal hyperalgesia were alleviated via intrathecal injection LV-hIL-10 in CCI rats. The overexpression of IL-10 were detected in spinal cord,pallium and hippocampus , especially in the spinal cord .Conclusions:The mechanical allodynia and thermal hyperalgesia can be relieved by intrathecal injection LV-hIL-10 in CCI rats.

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

OBJECTIVETo observe the effect of oxymatrine on the level of serum matrix metalloproteinase-2 (MMP-2) and its inhibitor (TIMP-2) in patients with chronic hepatitis B (CHB) and post-hepatitis B liver cirrhosis (LC), as well as on liver fibrosis indexes as hyaluronic acid (HA), laminin (LN) and IV type collagen (IV C).

METHODSChanges of all the above-mentioned indexes in patients with CHB (n = 36) and LC (n = 36) before and after treatment were determined, and the relationship of MMP-2 and TIMP-2 with liver fibrosis indexes were analyzed.

RESULTSOxymatrine could decrease the levels of MMP-2, HA, LN and IV C in patients with severe or moderate CHB and LC of Child-pugh A, B and C grade, as compared with the data before treatment (P < 0.05). Serum level of MMP-2 and TIMP-2 was well correlated with the levels of liver cirrhosis indexes.

CONCLUSIONMMP-2 and TIMP-2 can be used as a reference for evaluating the degree of LC, and oxymatrine has a certain anti-LC effect.

Adolescent ; Adult ; Aged ; Alkaloids ; therapeutic use ; Collagen Type IV ; blood ; Female ; Hepatitis B, Chronic ; complications ; drug therapy ; enzymology ; Humans ; Hyaluronic Acid ; blood ; Laminin ; blood ; Liver Cirrhosis ; complications ; drug therapy ; enzymology ; Male ; Matrix Metalloproteinase 2 ; blood ; Middle Aged ; Quinolizines ; therapeutic use ; Sophora ; Tissue Inhibitor of Metalloproteinase-2 ; blood

Objective: To explore the false-negative possibility in genetic test of congenital long QT syndrome (LQTS) by next-generation sequencing (NGS). Methods: A total of 28 genomic DNA samples were collected from 4 laboratories including 2 commercial medical laboratories using HiSeq2000 platform as Lab1,n=6 and Lab2,n=8; 1 commercial research service laboratory using Ion-torrent platform as Lab3,n=8 and 1 academic laboratory using HiSeq2000 platform as Lab 4,n=6. Sequencing coverage in the exons of protein-coding region in 3 main LQTS pathogenic genes as KCNQ1, KCNH2, SCN5A and possible pathogenic variants were quantitatively analyzed. Results: In Lab1, Lab 2 and Lab 4 with HiSeq2000 platform, above 98% protein coding regions in 3 pathogenic genes were covered with>10-fold reads and 90%-95% were covered with>30-fold reads. In 2 commercial medical laboratories, 3.63% and 9.84% protein coding regions of KCNQ1 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; lower than 10-fold covering region was focused in the 1st exon including about 2% known or likely pathogenic variants. In 2 commercial medical laboratories, 2.64% and 15.76% protein coding regions of KCNH2 gene in 14 samples were covered with<10-fold reads and with<30-fold reads; low covering region was located in multiple exons. For the data from Lab 1, as high as 28.56% protein coding regions of KCNH2 gene were covered with<30-fold reads including 113 (19.79%) known or likely pathogenic variants. SCN5A gene had the best coverage of protein coding region, with no<10-fold reads in all 4 Labs and no<30-fold reads in 2 commercial medical laboratories. Conclusion: Currently, NGS has low coverage region in both KCNQ1 and KCNH2 genes, pathogenic variants could be missed and false-negative possibility should be highly alert.

AIM:To investigate the characteristics of Ets-1 and VEGF expression and distribution in the experimental diabetic rat retina.METHODS:Diabetes was induced by intraperitoneal injection of streptozotocin (STZ).At 4 weeks after STZ-injection,animals were sacrificed.Total proteins were isolated from retinas of experimental and control eyes and were assessed by Western blot analysis.Frozen cross sections of eyeballs with 14um thickness were used to perform double immunoffuorescence staining with anti-Ets-1 and anti-VEGF antibodies.RESULTS:Both Ets-1 and VEGF expression were up-regulaled in the diabetic retina,the distribution of Ets-1 and VEGF was identical to each other,and the two proteins were almostlocalized in all retinal layers.CONCLUSION:Ets-1 might contribute to the pathologic progress of the diabetic retina induced by VEGF.

AIM: To determine the involvement of Ets-1 in the pathological progress of the experimental diabetic retina. METHODS: Diabetes was induced by intraperitoneal injection of STZ. Total RNA and Total proteins were isolated from retinas of experimental and control eyes at 4 weeks after STZ-injection and were assessed by Northern blot analysis and Western blot analysis, respectively.RESULTS: Expression of both Ets-1 mRNA and Ets-1 protein was significantly increased in the experimental diabetic rat's retina after STZ-injection compared with the control group (P＜0.001).CONCLUSION: Our results indicated that Ets-1 was involved in the pathological progress of experimental diabetic retina.Further studies should be conducted to focus on the relationship between Ets-1 and VEGF in the diabetic retina.

OBJECTIVETo investigate the expression of zinc ribbon domain-containing1 (ZNRD1) in human renal cell carcinoma and normal kidney tissues.

METHODSThe expression of ZNRD1 protein was examined by immunohistochemical staining in 71 renal cell carcinomas and 24 samples of normal kidney tissue. The correlation between the expressions of ZNRD1 protein and clinicopathologic features was analyzed. The expression of ZNRD1 mRNA and ZNRD1 protein was detected by quantitative reverse transcriptase-polymerase chain reaction (PT-PCR) and Western blot in 20 renal cell carcinomas and corresponding adjacent non-cancerous tissues.

RESULTSZNRD1 protein was detected mostly in the cell nuclei by immunohistochemistry. The positive expression rate of ZNRD1 protein was 91.7% (22/24) in renal cell carcinomas and 20.8% (5/24) in the normal kidney tissues, with a statistically significant difference between cancer and normal kidney tissue (P < 0.01). However, no significant correlation was observed between ZNRD1 protein expression level and clinicopathologic features (P > 0.05). ZNRD1 mRNA expression level was significantly higher in renal cell carcinomas (0.6186) than that in the normal kidney tissues (0.4273) assessed by RT-PCR (P < 0.01). The expression level of ZNRD1 protein by Western blot was 0.5623 in renal cell carcinomas, significantly higher than that in normal kidney tissues (0.3885, P < 0.01).

CONCLUSIONZNRD1 gene and ZNRD1 protein may play an important role in the carcinogenesis of renal cell carcinoma. Further investigation is still needed.

Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; metabolism ; pathology ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the change of proportion of peripheral blood dendritic cells (DCs) in patients with stroke. Methods 56 patients (30 cases of cerebral infarction and 26 cases of cerebral hemorrhage) in Beijing Bo'ai hospital from June to September, 2014 and 14 healthy controls were investigated. The severity of stroke was assessed with the National Institutes of Health Stroke Scale (NIHSS). Flow cy-tometry analysis was employed to detect the proportion of DCs subtypes in the peripheral blood. Results No obvious difference was found in DCs between the stroke patients and the controls. Compared to the control group, the percentages of peripheral blood myeloid dendritic cells (mDCs) decreased in the cerebral hemorrhage and the cerebral infarction subgroups (P<0.001). The percentages of plasmacytoid den-dritic cells (pDCs) reduced significantly in the cerebral hemorrhage and the cerebral infarction subgroups (P<0.05). The stroke patients were divided into NIHSS≤7 subgroup and NIHSS>7 subgroup. The percentages of pDCs in the cerebral hemorrhage and the cerebral infarction patients were significantly lower in the NIHSS>7 subgroup than in the NIHSS≤7 subgroup (P<0.05). While there was no statistical differ-ence between NIHSS≤7 subgroups and NIHSS>7 subgroups in the percentages of mDCs in the cerebral hemorrhage and cerebral infarction patients. Conclusion The proportion of DCs subtypes in the peripheral blood in stroke patients changed significantly, indicating inflamma-tion responds play a role in stroke.