· ATM:To analyze bacterial spectrum and drug sensitivity in conjunctival sac of cataract patients of Kazak.
· METHODS:A total of 538 cases of conjunctival sac secretion in cataract patients of Kazak were collected.The samples were cultured and their sensibilities to antibiotics were tested.
· RESULTS: The bacterial culture was positive in 214 cases.The positive rate was 39.8%. The variety of pathogenic bacteria were mainly made up of gram positive cocci ( 88.3%), and most of them were Staphylococcus epidermidis ( 66.4%), followed by Micrococcus(9.8%).Sex had no effect on conjunctival bacteria rate in the cataract patients of Kazak, while age, place of residence had an effect on camier rate. The camier rate of conjunctival bacteria was significantly higher in people over 60 years old than that in people with age between 40 to 59 years old.And the people from city had a significant lower bacteria positive rate than those from countryside and pastoral. Most of grams were sensitive to Vancomycin, Teicoplanin, Rifampicin, Duly cloth mildew mutual and Amikacin, the tolerance was less than 20%, and they usually had higher tolerance to Penicillin, Erythromycin, Tetracycline and Chloramphenicol (>70%) .
·CONCLUSlON:Gram positivecocci is the most common bacteria in conjunctival sac in cataract patients of Kazak. Staphylococcus epidermidis was most common, followed by Micrococcus.The germ-carrying rate of conjunctival SAC in Kazakh population is associated with the patient’s age and area of residence.The clinical use of antibacterial drugs should be strictly grasp the indications, to reduce the incidence of bacterial resistance.

OBJECTIVETo investigate the effect of ischemic postconditioning on the expression of rat myocardium matris metalloproteinase-2 (MMP-2) induced by ischemia/reperfusion (I/R) and relationship between its expression and interstitium and the effect on left ventricular function.

METHODSTwenty-four rats were randomly divided into 3 groups (n = 8): sham control (SC) group, ischemic/reperfusion (I/R) group and ischemic postconditioning (IPTC) group. The left ventricular peak systolic pressure and its derivate (+/- dp/dt) were calculated; The amount of myocardium collagenous were determined; The vitality of superoxide dismutase (SOD) and content of malondialdehyde (MDA) of plasma were detected; The activity of myocardium MMP-2 was measured by Western blot and RT-PCR.

RESULTSAs compared with I/R group, IPTC could lower the expression of MMP-2, ameliorate left ventricular function and increase the content of myocardium collagenous. In the meantime, the vitality of superoxide dismutase (SOD) of plasma were greatly enhanced and the content of malondialdehyde (MDA) of plasma were reduced in IFC group.

CONCLUSIONProtective effect of IPIC on myocardium may be due to reduce free radical, lower expression of MMP-2 and protect myocardial interstitium. MMPs plays an important role in the myocardial protection provided by IPTC.

Animals ; Collagen ; metabolism ; Ischemic Postconditioning ; Malondialdehyde ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Myocardial Reperfusion Injury ; enzymology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the effect of PC-1 gene knockdown on the biological action of prostate cancer cell line C4-2.

METHODSRecombinant plasmids of expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into C4-2 cells via liposome. The positive cell clones were selected by G418. The expression of PC-1 gene was analyzed by RT-PCR and Western blotting technology. MTT and soft agar cloning formation were applied to observe the changes of the growth rate and independent anchor ability of C4-2 cells.

RESULTSPC-1 RNA interference severely affected the expression of PC-1 gene and reduced the growth and colony formation ability of C4-2 cells.

CONCLUSIONRNA interference-mediated PC-1 gene knockdown can decrease the growth and cloning formation ability of C4-2 cells.

Cell Line, Tumor ; Down-Regulation ; Gene Expression ; Humans ; Male ; Phosphoric Diester Hydrolases ; biosynthesis ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Pyrophosphatases ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo observe the effect of p21-activated kinase-1 (PAK1) gene transfection on the invasiveness of human colorectal carcinoma SW480 cells in vitro.

METHODSSW480 cells in routine culture were transfected with the recombinant plasmid EGFP-C1/PAK1 via Lipofectamine(TM) 2000. The expression of PAK1 protein in SW480 cells was detected using Western blotting, and the changes of the invasiveness of SW480 cells were evaluated using Boyden chamber invasion assay.

RESULTSForty-eight hours after transfection with pEGFP-C1/ PAK1, the PAK1 protein expression increased significantly in comparison with those in negative and vector control groups. The invasiveness of the SW480 cells was significantly enhanced after the transfection.

CONCLUSIONThe PAK1 gene transfection can increase the expression of PAK1 in SW480 cells and enhance the invasiveness of the cells. PAK1 can be associated with the invasiveness and metastasis of colorectal carcinoma cells.

Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; pathology ; Gene Expression ; Genetic Vectors ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Plasmids ; Transfection ; p21-Activated Kinases ; genetics

OBJECTIVETo investigate the effect of short hairpin RNA (shRNA) targeting survivin on adhesion and invasion of human colon carcinoma cell line SW480 in vitro.

METHODSAccording to the sequence of the coding region of survivin gene, two strings of 19 nucleotides of inverted sequence flanking the loop sequence of two complementary 9-base oligonucleotides were designed and synthesized to prepare the hairpin construct as the DNA templates for the target shRNA. The shRNA templates were cloned into shRNA expression vector pRNAT-U6.1/Neo, and the resulted vector pRNAT-U6.1/Neo-survivin was transfected into SW480 cells using Lipofectamine 2000. Western blotting was performed to evaluate survivin gene silencing induced by shRNA transfection at the protein level, and the biological behaviors of the SW480 cells were investigated by cell-matrix adhesion, invasion and gelatin-zymography assays.

RESULTSWestern blotting revealed significantly lowered survivin protein expression in transfected SW480 cells, and survivin gene silencing induced by shRNA significantly suppressed the metastatic potential of SW480 cells in association with suppressed MMPs activity.

CONCLUSIONSSurvivin may play an important role in modulating human colorectal carcinoma cell invasion and metastasis, and survivin gene silencing can inhibit human colorectal cancer cell invasion and the production of MMP-2 and MMP-9. Survivin may affect invasion and metastasis of human colorectal carcinoma cells via regulating the production of MMPs.

Animals ; Blotting, Western ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; pathology ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; Inverted Repeat Sequences ; Matrix Metalloproteinases ; secretion ; Microtubule-Associated Proteins ; deficiency ; genetics ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics ; RNA, Small Interfering ; genetics

Objective To study the HIV-1 drug resistance (DR) situation among newly infected persons in Dehong.Methods 1048 HIV-1 positive blood samples from July to December in 2006 from Dehong prefecture of Yunnan,were collected.HIV drug resistance were tested using TruGene in newly infected people that were distinguished with BED-CEIA,while the subtype were determined with phylogenetic analysis using a set of reference sequences available on the Los Alamos Database.Results Of sixty-four successfully analyzed samples,drug resistance mutations were detected in 4 samples with the resistance rate as 6.25%.Minor mutation in PR region such as M36I/V,L63P and H69K appeared frequently and the rates were 81.25%,70.31%and 65.63%respectively.The predominantly prevalent strains were seen as C/CRF07_BC/08_BC(65.63%,42/64) in this study.Conclusion The prevalence of genotypic drug resistances in HIV-1 recent infections in Dehong prefecture appeared to be at moderate level.Drug-resistance surveillance program among HIV-1 infections should be continued and strengthened.

OBJECTIVETo investigate the relationship between the dendritic cell (DC) subsets and transcriptive factors, T-bet, GATA-3, and immune imbalance in acquired severe aplastic anemia (SAA).

METHODSThe DC1 (HLA-DR+Lin-CD11c+) and DC2 (HLA-DR+Lin-CD123+) in peripheral blood mononuclear cells (PBMNC) were measured with flow cytometry (FCM), the expressions of T-bet mRNA and GATA-3 mRNA in PBMNC with semiquantitative RT-PCR and the plasma level of IFN gamma and IL-4 with ELISA in 29 SAA patients and 16 healthy controls.

RESULTSThe percentages of DC1 in PBMNC were (0.44 +/- 0.24)% and (0.73 +/- 0.30)% in untreated and recovered SAA patients respectively, both were higher than that in controls (0.29 +/- 0.10)% (P < 0.05). The percentage of DC2 in the untreated cases was lower than that of recovered ones or controls [(0.18 +/- 0.14)% vs (0.28 +/- 0.20)% and (0.29 +/- 0.13)%] (P < 0.05). DC1/DC2 ratios were 3.45 +/- 2.71 and 2.90 +/- 0.95 in untreated and recovered groups respectively, both were higher than that in controls (1.15 +/- 0.56) (P < 0.05). No statistic difference in DC1/DC2 ratio was found between untreated and recovered patients (P < 0.05). The relative mRNA expression levels of transcriptive factor T-bet were 0.37 +/- 0.07, 0.20 +/- 0.07 and 0.17 +/- 0.05 in the above 3 groups, respectively, untreated group being higher than that of recovered group or healthy controls (P < 0.05). There was no statistic difference of GATA-3 expression among the 3 groups (P > 0.05). T-bet/GATA-3 ratio was 0.72 +/- 0.13 in untreated group, being higher than that of recovered group (0.33 +/- 0.08) or controls (0.35 +/- 0.11). The plasma level of IFN gamma in the untreated group was (50.9 +/- 1.1) ng/L, which was higher than that of recovered group [(49.7 +/- 0.9) ng/L] or controls [(49.7 +/- 0.7) ng/L]. There was significant positive correlations between T-bet and DC1/DC2 ratio (r = 0.445, P < 0.01), as well as between T-bet and IFN gamma (r = 0.402, P < 0.01).

CONCLUSIONEither DC1/DC2 or T-bet/GATA-3 ratio might become an index to estimate immune imbalance. High-expressed T-bet was related to the progress of SAA. In patients with SAA, DC1/DC2 ratio returns to normal range later than that of routine blood test does, indicating that immunosuppressive therapy should not be withdrawn too earlier.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; immunology ; Child ; Dendritic Cells ; immunology ; Female ; GATA3 Transcription Factor ; blood ; genetics ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; RNA, Messenger ; genetics ; T-Box Domain Proteins ; blood ; genetics ; Young Adult

This study was aimed to investigate the quantity and subtypes of dendritic cells (DC) in patients with immune related pancytopenia (IRP) and to explore the role of DC in pathogenesis of IRP. The quantity of plasmacytoid dendritic cells (pDC, Lin(-)HLA-DR(+) CD123(+) cells) and myeloid dendritic cells (mDC, Lin(-)HLA-DR(+) CD11c(+)cells) in peripheral blood of 65 patients with IRP (37 new diagnosed and 28 remitted) and 17 healthy controls were analyzed by flow cytometry. The results indicated that the ratio of pDC in peripheral blood mononuclear cells (PBMNC) was (0.91 ± 064)% in new diagnosed group, which was significantly higher than that in remission group (0.39 ± 0.11)% and control group (0.29 ± 0.13)% (P < 0.01), while this ratio of pDC in remission group was higher than that in control group (P < 0.05). The ratio of mDC in PBMNC was (0.21 ± 0.20)% in new diagnosed group and (0.34 ± 0.21)% in remission group respectively, there was no statistical difference as compared with control group (0.29 ± 0.09)% (P > 0.05). The ratio of pDC to mDC in new diagnosed group was 6.75 ± 7.11, which was significantly higher than that in remission group (1.55 ± 0.93) and control group (1.07 ± 0.43, P < 0.01), there was no statistical difference between the ratio of remission group and control group (P > 0.05). The ratio of pDC in PBMNC of IRP group negatively correlated to ratio of Th1/Th2 (r = -0.347, P < 0.05), and positively correlated to the ratio of auto-antibody on membrane of BMMNC (r = 0.606, P < 0.05) and to the quantity of CD5(+)B cells (r = 0.709, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.381, P < 0.01) and platelets (r = -0.343, P < 0.01). The ratio of mDC in PBMNC positively correlated to the ratio of Th1/Th2 (r = 0.595, P < 0.05) and the level of hemoglobin (r = 0.292, P < 0.05). The ratio of pDC/mDC negatively correlated to ratio of Th1/Th2 (r = -0.395, P < 0.05), it positively correlated to the level of antibody on membrane of BMMNC (r = 0.421, P < 0.05) and the quantity of CD5(+)B cells (r = 0.423, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.304, P < 0.05) and platelets (r = -0.287, P < 0.05). It is concluded that the quantity of pDC in peripheral blood of IRP patients increases, which may be related to the immunopathogenesis of IRP.
Adolescent ; Adult ; Blood Cell Count ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; cytology ; immunology ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; immunology ; Young Adult

This study was purposed to applicate the bone marrow indirect Coombs test and investigate its clinical significancies in diagnosis of immuno-related pancytopenia (IRP). 30 patients with pancytopenia including 22 cases of IRP and 8 cases of idiopathic cytopenia of undetermined significance (ICUS), and 15 patients with iron-deficiency anemia as controls were enrolled in this study. After incubation of the bone marrow supernatant of IRP patients and bone marrow nucleated cell (BMNC) of controls was used as experiment group, while the incubation of BMNC and bone marrow supernatant of controls was used as control group. After incubation for 45 min, the positive rate of membrane antibodies in bone marrow hematopoietic cells (CD15(+), GlyCoA(+) and CD34(+)cells) was detected by flow cytometry, and correlation analysis of positive rate with clinical data of patients were analyzed. The results showed that among 30 patients with pancytopenia (16 positive and 14 negative for bone marrow direct Coombs test) 16 cases showed positive for bone marrow indirect Coombs test, with positive rate 53.33. In the experiment group, the median positive rate of CD15(+)IgM was 0.34, which was significantly higher than that in control group (0.20, P < 0.05); the median positive rates of CD34(+) IgG and IgM were 0.64 and 0.21 respectively, which were significantly higher than those in control group (0.00, P < 0.05) and (0.00, P < 0.05); the positive rates of GlyCoA(+)IgG and IgM were (0.83 ± 0.75) and (2.12 ± 1.98) respectively, which were significantly higher than those in control group [(0.47 ± 0.43), P < 0.05, (0.68 ± 0.64), P < 0.01]; the positive rates of CD15(+) IgG and IgM were positively correlated with the ratio of CD5(+)B cells. The positive rates of GlyCoA(+) IgG and IgM negatively correlated with the Hb level, percentage of reticulocytes, the ratio of bone marrow erythroid lineage and DC1/DC2 positively correlated with the ratio of CD5(+)B cells and indirect bilirubin level. It is concluded that antibodies (IgG or IgM) aiming at the bone marrow hematopoietic cells exist in the supernatant of some IRP and ICUS patients, and may act on the membrane protein of the normal BMNC. These antibodies correlate with the prognosis of IRP.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; immunology ; Child ; Coombs Test ; Female ; Humans ; Male ; Middle Aged ; Pancytopenia ; diagnosis ; immunology ; Young Adult

The goal of treatment of metabolic syndrome is the prevention of diabetes and cardiovascular events. A series of novel tetrahydrocoptisine quaternary ammonium compounds were prepared to evaluate their action of hypoglycemia and hypolipidemia for finding the therapeutic agents of metabolic syndrome. Starting from the coptisine hydrochloride (2), fifteen target compounds were synthesized by reduction and substitution of the 7-N position. All of the target compounds were characterized by 1H NMR and HR-MS. Their hypoglycemic activities were evaluated in HepG2 cell and hypolipidemic activities of compounds with better hypoglycemic activity were tested further in vivo. Results indicated that compounds 5, 7, 8 and 9 exhibited better hypoglycemic activities in vitro and compounds 5 and 8 exhibited good hypolipidemic activities in high-fat-diet (HFD) induced hyperlipidemia mice and (or) hamsters. However, the activity is not as good as simvastatin.
Animals ; Berberine Alkaloids ; chemical synthesis ; chemistry ; pharmacology ; Cholesterol ; blood ; Glucose ; metabolism ; Hep G2 Cells ; Humans ; Hyperlipidemias ; blood ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Hypolipidemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Mesocricetus ; Mice ; Molecular Structure ; Quaternary Ammonium Compounds ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Triglycerides ; blood