Multidrug resistant genes are highly expressed in hepatocellular carcinoma that seriousty affects the effect of chemotherapy. Screening of resistant genes from HCC cells and studying its mechanism of drug resistance will be helpful to improve the effecacy of chemotherapy for hepatocellular carcinoma. Here we described an alternative method called cyclical packaging rescue (CPR). First we constructed a retrovirus cDNA library of hepatoma cells and used it to infect fibroblasts. Then we added drugs to screen survival cells. The survival cells, stably integrated helper-free retroviral libraries, were recovered rapidly after transfection with plasmids expressing retroviral gag-pol and env genes. Through this method, retroviral RNAs were directly repackaged into new infectious virions. Recovered retroviral supernatant was then used to reinfect fresh target cells. When performed in concert with selection using functional assays, cDNAs regulating functional responses could be identified by enrichment through multiple rounds of retroviral library recovery and retransmission. Using CPR, we obtained several cDNAs. After a preliminary detection, we found Ribosomal protein S11 (RPS11), Ribosomal protein L6 (RPL6), Ribosomal protein L11 (RPL11), Ribosomal protein L24 (RPL24) possibly had drug resistant function.
Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; DNA, Complementary ; Drug Resistance, Neoplasm ; genetics ; Gene Library ; Genetic Vectors ; Humans ; Liver Neoplasms ; genetics ; pathology ; Plasmids ; Retroviridae ; Ribosomal Proteins ; genetics ; metabolism ; Transfection

Objective To study ischemic tolerance induced by acanthopannx senticosus saponins(ASS) on ischemia in PC12 cells and involve expression of hypoxia inducible factor-1?(HIF-1?).Methods An ischemic model was developed in PC12 cell line with treatment of oxygen glucose deprivation(OGD).The protective effects of ASS pretreatment on ischemic tolerance of PC12 cells and whether protective effects of ASS could be inhibited by LY294002(PI3K inhibitor) were analyzed through MTT assay.The induction of phospho-glycogen synthesis kinase-3?(p-GSK-3?) and HIF-1? after pretreatment of ASS and possible blocking effects of LY294002 were detected by Western-blot.Results MTT results showed that after 9 h ischemia,the viability of PC12 cells decreased dramatically(P

Objective To evaluate the distribution and drug resistance of bacteria causing neonatal infectious pneumonia in Jiaxing,and to provide a therapy for clinical doctor to make a correct diagnosis,choose reasonable anti-biotics and avoid abuse of antibiotics.Methods Took expectoration from trachea in condition of asepsis to conduct culture and perform drug-sensitive test from 3025 cases.Results Totally 1 156 strains of aerobic bacteria were iso-lated.875 strains were gram negative bacilli(75.7%),269 strains were gram positive cocci(23.3%),and 12 strains were fungi(1.0%).Klebsiella pneumoiae,Escherichia coil,Acinetobacter baumanni,Enterobacter cloacae were com-mon in gram negative bacilli( respectively 178 cases,151 cases,87 cases,113 cases) .The proportion of the Staphylo-coccus aureus was the largest in gram positive cocci(245 cases) .The results showed that gram-negative bacilli were resistant to cefazolin, ampicillin, piperacillin and sensitive to meropenem, imipenem, piperacillin-tazuobatanna and cefoperazone-sulbactam.The drug resistance was severe of ESBL-positive.Staphylococcus aureus was resistant to penicillin, ampicillin, erythrocin, clindamycin and sensitive to linezolid, vancomycin, nitrofurantoin.Conclusion Gram-negative bacilli are the main bacteria in neonatals with infectious pneumonia.The drug resistance is severe.It is important to make a standard management and isolation.

Objective To explore antibiotics resistance profiles and DNA fingerprints of Pseudomonas aeruginosa isolates resistant to imipenem (IRPA). Methods DNA fingerprints of 56 strains isolated from ICU (intensive care unit) were constructed by ERIC-PCR (enterobacter repetitive intergenic consensus-PCR). MICs (minimal inhibitory concentrations) were determined by agar dilution method.Results 33 genotypes were got from 56 strains by ERIC-PCR. Of 8 frequently used antibiotics, 5 of them showed resistance rate higher than 50%. Conclusion It is high time to pay attention to multi-drug resistance of IRPA. The exist of prevalence of IRPA clone in ICU advise us to control of IRPA in hospital effectively.

0.05). On the day4, day5, the expression of E-selectin protein were greatly higher than that of normal endometrium (57.52 + 7.03, 62.91 + 7.31; P

Objective To investigate the imaging characters of abdominal cocoon.Methods Six cases of abdominal cocoon proved by surgery and pathologic findings were retrospectively analyzed. Abdominal plain X-ray and CT were performed in 6 cases.The gastrointestinal barium meal series were undergone in 4 cases.The imaging findings were analyzed.Results Abdominal plain X-ray suggested intestinal obstruction in 3 of 6 cases.The gastrointestinal barium meal showed"cauliflower sign"or "concertina pattern"in all of the 4 cases;CT images revealed a conglomeration of multiple small bowel loops in all 6 cases and the intestinal loops seemed to be encapsulated in a membranelike sac.Conclusion The imaging features of gastrointestinal barium meal and CT scan could suggest the diagnose of abdominal cocoon.

OBJECTIVETo investigate the clinical effectiveness of sertraline hydrochloride combined with four-spot caress in the treatment of primary premature ejaculation (PE).

METHODSWe randomly assigned 90 primary PE patients to three groups of equal number. The patients in group A (aged [28.1 ± 5.2] yr and with a disease course of [3.1 ± 1.9] yr) were treated with oral sertraline hydrochloride at 50 mg qd, those in B (aged [27.8 ± 4.1] yr and with a disease course of [3.2 ± 2.0] yr) by four-spot caressing (caressing the tongue, breasts, and vulva prior to intercourse), and those in C (aged [27.1 ± 4.7] yr and with a disease course of [3.1 ± 2.0] yr) by the combination of oral sertraline hydrochloride and four-spot caressing, all for 12 weeks. Before and after 4, 8, and 12 weeks of treatment, we obtained the intravaginal ejaculatory latency time (IELT) and Chinese Index of Sexual Function for Premature Ejaculation-5 (CIPE-5) scores and compared them among the three groups of patients.

RESULTSThe IELT was dramatically prolonged in groups A, B, and C after 4 weeks ([1.08 ± 0.29], [0.93 ± 0.28] and [1.21 ± 0.27] min), 8 weeks ([1.43 ± 0.30], [1.20 ± 0.33] and [1.72 ± 0.42] min) and 12 weeks of treatment ([2.12 ± 0.63], [1.90 ± 0.65] and [2.67 ± 0.82] min) as compared with the baseline ([0.63 ?0.14] , [0.60 ?0.14] and [0.62 ?0.11] min) (P < 0.05), even longer in group C than in A and B (P < 0.05). The CIPE-5 scores were markedly improved in groups A, B and C after 4 weeks ([15.17 ± 1.74], [14.57 ± 1.94] and [15.60 ± 1.63] min), 8 weeks ([17.13 ± 1.63], [16.37 ± 1.97] and [18.00 ± 1.05] min) and 12 weeks of intervention ([18.93 ± 1.57], [18.53 ± 1.67] and [20.00 ± 1.46] min ) as compared with the baseline ([12.57 ± 2.05], [13.20 ± 2.51] and [13.07 ± 2.01] min) (P < 0.05), even higher in group C than in A and B (P < 0.05).

CONCLUSIONSertraline hydrochloride combined with four-spot caressing, with its definite efficacy and rare adverse reactions, deserves wide clinical application in the treatment of primary PE.

Adult ; Coitus ; Ejaculation ; Female ; Humans ; Male ; Premature Ejaculation ; drug therapy ; Sertraline ; therapeutic use ; Young Adult

To explore the effect of tripterygium glycosides on the level of peripheral blood cell factors of Graves ophthalmopathy (GO). In the study, 64 patients of GO in moderate-severe acute stage were selected, and randomly divided into the treatment group (32 cases) and the control group (32 cases). Both of the two groups were provided with basic treatment. The control group was added with prednisone(0. 75 mg kg-1 d-1 ), which gradually reduced (by 5-10 mg week-1 )to the minimum dose of 5 mg d-1. The treatment group was treated with 20 mg tripterygium glycosides, three times a day. One therapy course is three months. The levels of peripheral blood cells(TNF-alpha , IL-2, IL-10, IFN-gamma)of the two groups before and after the treatment and the clinical efficacy were observed. The study indicated that, before the treatment, TNF-alpha, IL-2, IFN-gamma in both groups were significantly higher than that in the health group, but with IL-10 notably lower than the healthy group. After the treatment, TNF-a, IL-2, IFN-gamma in the treatment group significantly decreased, but with IL-10 significantly increasing (P <0. 01). After the treatment, the two groups showed significant difference (P <0. 01). The total clinical efficacy in the treatment group was 88. 10% , and that in the control group was 57. 14% (P <0. 01). After the treatment, the two groups showed significant changes in the exophthalmos degree (P < 0. 01). The results showed that the level of peripheral blood cells (TNF-alpha,IL-2, IL-10, IFN-gamma)of GO patients was positively correlated with the severity of ocular disease. The combined therapy of tripterygium glycosides and methimazole show such advantages as low side effect and high clin-
Adult ; Cytokines ; blood ; Female ; Glycosides ; pharmacology ; therapeutic use ; Graves Ophthalmopathy ; blood ; drug therapy ; Humans ; Male ; Tripterygium ; chemistry

OBJECTIVETo study the tolerance to ischemia induced by Acanthopanax Senticosus Saponins (ASE) in PC12 cells and the involved mechanism.

METHODSAn ischemic model was developed in PC12 cell line by treatment with oxygen-glucose deprivation. The effects of ASE pretreatment on tolerance of PC12 cells to ischemia were evaluated by MTT assay and analysis of cellular morphology. The expression of hypoxia-inducing factor (HIF)-1alpha, erythropoietin (EPO) after the pretreatment with ASE was detected by Western blotting. The DNA binding activities of HIF-1 in PC12 cells with the pretreatment of ASE were demonstrated by using electrophoretic mobility shift assay (EMSA).

RESULTSIn ischemia model, the viability of PC12 cells was decreased to (49.12 +/- 3.22)% after oxygen-glucose deprivation for 9 hours. However, ASE (50 microg/ml) pretreatment could remarkably increase the viability of PC12 cells by (67.97 +/- 2.92)%. There were significant differences between the experimental group and control group (F = 473.67, P < 0.01). The cellular morphology showed that PC12 cells exposed for 7 days to nerve growth factor (NGF) exhibited round, smooth cell bodies with normal processes and that processes formed extensive network. At 9 hour after ischemia, cell bodies of many PC12 cells were found shrinken, the processes were disrupted and network disappeared. However, pretreatment with ASE (50 microg/ml) could largely prevent the morphological damage to PC12 cells that would have caused by subsequent exposure to 9 h ischemic insult, many cellular bodies were intact and many processes and network of PC12 cells still existed. The expression of HIF-1alpha increased after pretreatment with ASE shown by Western blot. There were significant differences between the experimental group and control group (F = 167.18, P < 0.01). The DNA binding activities of HIF-1 in PC12 cells after pretreatment with ASE was significantly increased, and it could activate the expression of EPO (F = 128.37, P < 0.01).

CONCLUSIONSThe pretreatment with ASE could induce tolerance against ischemia in PC12 cells. The elevated expression and increased DNA binding activity of HIF-1alpha, the overexpression of its downstream target EPO may be the molecular mechanism in tolerance of PC12 cells to ischemia induced by ASE pretreatment.

Animals ; Cell Hypoxia ; Cell Survival ; Electrophoretic Mobility Shift Assay ; Eleutherococcus ; chemistry ; Erythropoietin ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Ischemia ; prevention & control ; PC12 Cells ; Rats ; Saponins ; pharmacology

Objective To screen differentially expressed serum proteins by label-free quantitative proteomics in patients with atopic dermatitis,in order to discover serum protein markers for atopic dermatitis.Methods Serum samples were collected from 6 patients with atopic dermatitis and 6 healthy human controls.Then,the samples from the patients were pooled together,so with those from the controls.Proteins were extracted from the two joint serum samples followed by separation with one-dimensional sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE).Peptides were obtained by enzymolysis in protein gels,which were subsequently identified by high performance liquid chromatography tandem mass spectrometry.The Mascot software was used for database searching,and the Scaffold software for relatively quantitative analysis of proteins.Results Totally,76 proteins with a difference of greater than 1.5 folds were identified between the serum samples of patients with atopic dermatitis and controls.There were 50 up-regulated and 26 down-regulated serum proteins in the patients compared with the healthy controls.Among these differentially expressed proteins,26 were only detected in the sera of the patients,and 14 proteins in those of the controls.Conlusion Differences exist in the expression of serum proteins between atopic dermatitis patients and healthy people.