We used MTT assay to test the cellular cytotoxicity ( NK, LAK, CTL, Macrophage), cytokine activities ( 1L-1, 1L-2, 1L-6, TNF), proliferation of lymphocytes and chemosensitivity of tumor cells, and compared it with radioactive isotope assay. The results showed that the MTT assay may be used to test the cellular cytotoxicity, cytokine ac-tivity, proliferation of lymphocytes and chemosensitivity of tumor cells. We think it is a simple, rapid, economic and safety method.

As a disease difficult to treat,the refractory ascites due to post hepatitis often results in a bad prognosis because the general medical treatment is ineffective.This paper introduces the current interventional treatment of the refractory ascites.

Objective To explore the diameter change of the extrahepatic bile duct before and after laparoscopic cholecystectomy (LC). Methods From Jan. 2006 to Dec. 2007, 113 patients including chronic gallstone cholecystitis (n=55), inactive cholecystolithiasis (n=46) and gallbladder polyps (n=12) were collected and treated by LC. The diameters of their extrahepatic bile ducts were measured by B ultrasonography before operation, 3 months and 6 months after operation. These data were collected and analyzed retrospectively. Results The diameters of the extrahepatic bile ducts of all patients before LC, 3 months and 6 months after LC were (5?2) mm, (8?2) mm and (6?2) mm respectively. And in chronic gallstone cholecystitis patients they were (5?2) mm, (9?2) mm and (6?2) mm respectively, in inactive gallstone cholelithiasis patients they were (5?2) mm, (8?2) mm and (6?2) mm respectively, and in gallbladder polyps ones they were (5?2) mm, (7?2) mm and (5?2) mm respectively. Conclusion The change of the extrahepatic bile duct diameter after LC is a dynamic process. It is enlarged on the third month after operation than before operation. In the sixth month after operation marked retraction occurs, and compared with before operation, it shows no obvious statistic significance.

Objective To investigate the alternations in gene/amino acid sequence of penicillin-binding protein (PBP) 2b from clinical isolates of penicillin-nonsusceptible Streptococcus pneumonia(PNSP) in this region.Methods 24 strains of Streptococcus pneumonia were collected from January to December 2006.The antibiotics susceptibility of these strains was detected.PCR amplification and direct sequencing of pbp2b genes were performed.The sequence variations of PBP genes of the PNSP in this region were studied with sequence BLAST analysis.Results Three prominent substitutions were common tO 13 PNSP isolates with minimal inhibitory concentration(MIC) at least 0.1 mg/L.These included the replacement of Thr445→Ala following the conservative motif SSN,Glu475→Gly and Thr488→Ala/Ser.The exchange of Glu332→Gly was identified in 12 PNSP isolates of which the MIC was at least 0.25 mg/L.Seven penicillin resistant Streptococcus pneumonia (PRSP) isolates (MIC≥3 mg/L)shared the amino acid substitution Ala618→Gly adiacent to third conserved (KTG) motif and the PBP2b sequences of seven PRSP isolates were classified within Back's group Ⅱ and were very similar to those of the Korean J77 isolate.Novel gene and amino acid sequence variants in isolate 14,15,8,11 and 24 was identified in this study and these gene sequences have been deposited in the GenBank database and assigned accession no.EU035970,EU056919,EU056920,EU056921 and EU106886.Conclusion Analysis of pbp2b genes revealed highly similar patterns of nucleotide and amino acid sequence variation among most resistant isolates.while penicillin intermediate Streptococcus pneumonia might be associated with novel gene sequence variants.

Objective To investigate the association of T help (Th) lymphocyte and heart rate variability (HRV) in congestive heart failure (CHF) patients. Methods Ninety-six patients with CHF and thirty healthy persons were enrolled in the study. Time-domain HRV analysis was performed based on 24 hour Holter Electrocardiogram (ECG) monitoring. Interferon-γ (IFN-γ) was used as markers for the differentiation of Th1 subsets and interleukin-10 (IL-10) for the Th2 subsets. IFN-γand IL-10 in CD4 + T lymphocytes were quantified by 3-color flow cytometry. Results The frequency of IL-10-Producing T Cells in the CHF group was significantly lower than those in the healthy control ( ( 16.4 ± 5.8 ) % vs. ( 26.8 ± 3.7 ) %, t = 9. 243, P ＜ 0. 001 ). The frequency of IFN-γ in the CHF group ( ( 18.4 ± 7.3 )% ) was significantly lower than that in the healthy controls ( (7.3 ±4.6) % ,t =7. 917, P ＜ 0. 001 ). The following index of HRV in the CHF groups were all significantly lower than those in the healthy controls: (98. 6 ± 21.3) ms vs. ( 145. 1 ± 42. 6) ms for SDNN, (83. 9 ± 22.4) ms vs.(136.5 ±39.6)ms for SDANN, (40.6 ± 14.5) ms vs. (55.8 ± 17.9) ms for SDNNI, (20. 7 ± 12.9) ms vs.(29.1 ± 12.6) ms for RMSSD, (5.6 ± 3.7 ) % vs. ( 11.8 ± 4.4) % for PNN50 ( Ps ＜ 0.05 ). In CHF patients, the frequency of IL-10 were positively associated with SDNN, SDANN, SDNNI, RMSSD and PNN50 ( r = 0. 49,0. 57,0. 58,0.47 and 0. 52 ,respectively,Ps ＜ 0.01 ). In the CHF patients, the frequency of IFN-γand IFN-γ/IL-10 ratio were negatively associated with SDNN ,SDANN ,SDNNI, RMSSD and PNN50 ( r = - 0. 49, - 0. 54, - 0. 57, - 0.52,- 0.53, - 0. 52, - 0.64, - 0.57, - 0. 58, - 0. 67, Ps ＜ 0. 01 ). Conclusions Autonomic nervous system is involved in the regulation of the balance of Th1/Th2 in patients with CHF. Sympathetic nerve system enhances the effect of Th1 ,whereas parasympathetic nervous system enhances the effect of Th2.

AIM: To study rhIL-1? effects on fetal islet function and IL-6 production in vitro METHODS: Islets from fetal pancreas was separated by collagenase type V (0 5 mg/mL) and cultured in vitro The islets were exposed to culture medium alone for 48 h or with different concentration of rhIL-1? The supernatants of culture of human fetal islets were assayed for IL-6, insulin and glucagon RESULTS:(1) IL-6 activity was increased 4 0 folds (74-294 mU/islet) when islets were exposed to rhIL-1?(20U/mL); (2) IL-6 McAb significantly reduced IL-6 activity in islet supernatants from control group or islet exposed to rhIL-1? treated group; (3)IL-6 mRNA in human fetal islet exposed to rhIL-1? is higher than control in dot hybridization; (4) Soluble insulin and cellular insulin within islet released to supernatants was slightly decreased (0 48～0 78 IU/islet and 0 65～0 79 IU/islet); (5) Glucagon secretion was significantly increased 3 2 folds (1 0～3 2 pg/islet) CONCLUSION: Pancreatic islets produce IL-6 is up-regulated by rhIL-1? On the other hand, Il-6 produced by the islet may act as a costimulator for autoreactive B and T lymphocytes in autoimmune diabetes

OBJECTIVE:To investigate the protective effects of insulin-glucose on myocardium in patients receiving cardiac valve replacement under cardiopulmonary bypass. METHODS:Totally 120 patients receiving combined cardiac valve replacement under cardiopulmonary bypass were divided into control group and observation group according to random number table,with 60 cases in each group. All patients were given routine operation. Control group was given Thomas cardioplegia and oxygenated blood with a ratio of 1:4(V:V)to protect myocardium at 4 ℃. Besides that,the observation group was additionally given Insulin injec-tion 10 IU/L and Glucose injection 10 g/L added into Thomas cardioplegia at 4 ℃ to protect myocardium. The levels of plasma brain natriuretic peptide(BNP)and cardiac troponinⅠ(cTnⅠ)before anesthesia induction(T0),at the end of cardiopulmonary by-pass(T1),12 h(T2),24 h(T3),48 h(T4),and 72 h(T5)after surgery,the rate of recovery of automatic heartbeat after opening aor-ta,the application of vasoactive agent(dopamine)at T1 and the occurrence of postoperative complications were observed and com-pared between 2 groups. RESULTS:At T0,there was no statistical significance in the levels of plasma BNP and cTnⅠ between 2 groups(P>0.05). The levels of plasma BNP and cTnⅠin 2 groups at T1-5 were significantly higher than T0,with statistical signifi-cance(P<0.05);the levels of cTnⅠ began to decrease at T4 and cTnⅠbegan to decrease at T5. However,the levels of BNP and cTnⅠwere significantly lower in observation group than in control group at T1-5,with statistical significance(P<0.05). After open-ing aorta,there was no statistical significance in the rate of recovery of automatic heartbeat between 2 groups(P>0.05). The dos-age of dopamine (at T1) and the incidence of complications in observation group were statistically lower than control group,with statistical significance(P<0.05). No severe ADR was found in 2 groups during or after surgery. CONCLUSIONS:Insulin-glucose can alleviate myocardial damage, reduce the dosage of vasoactive agent and the incidence of postoperative complications in pa-tients receiving combined cardiac valve replacement under cardiopulmonary bypass with significant protective effect on myocardium with good safety.

Objective To construct and screen effective shRNA expression vectors targeting human AQP1 gene ,and evaluate the interference efficiency of the AQP1 shRNA recombinant plasmids ,thus provide basis for further exploration on the effect and mechanism of AQP1 gene on human breast cancer cells .Methods Four pairs of shRNA sequences targeting human AQP1 gene were designed and synthesized ,and then inserted into the GV115 vector .AQP1 shRNA and control shRNA plasmids were trans‐fected into human breast cancer MCF‐7 cells .The expression of AQP1 mRNA and protein were detected by real time PCR(RT‐PCR) and Western blot to evaluate the interfering efficiency .Results RT‐PCR demonstrated that AQP1 was expressed in human breast cancer MCF‐7 cells .Sequencing showed that the shRNA vectors targeting AQP1 were successfully constructed .48 h after the AQP1 shRNA transfection ,AQP1 mRNA and protein expression levels in MCF‐7 cells were reduced to a significant degree ,and the AQP1 shRNA 4 plasmid vector could inhibit the AQP1 most efficiently .Conclusion The AQP1 shRNA recombinant plasmids vectors were successfully constructed and can significantly inhibit the expression of AQP1 in MCF‐7 human breast cancer cells .

Objective: To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of cord blood CD34+ cells. Methods: Human cord blood mononuclear cells (MNC) were cultured with different combinations of HGFs (including rhIL-1,rhIL-3, rhIL-6, rhG-CSF, rhGM-CSF and rhSCF). The expansion folds of MNC, the changes of cellular surface markers by FACS, and CFU-GM of hematopoietic cells were observed. Results: The total nucleated cells expanded by 44 folds after cultured with 6 cytokines for 20 days. The number of CFU-GM increased by 14 .74 folds after cultured in liquid for 8 days, but decreased heavily after 18 days. The total number of CD34+ cells increased by 127. 79 ~ 196 .40 folds at 6 to 8 day, but decreased to 101.51 folds at 18 day. Conclusion: Human hematopoietic growth factors can increase the expansion of cord blood CD34+ cells and CFU-GM significantly.

Objective:To study the characterization of NK cell line co-transfected with hSCF and hIL-15 gene.Methods:pcDNA3-hSCF and pcDNA3-hIL-15 were transfected into NK-92 cell line with LipofectAMINETM,RT-PCR was used to identify NK-92 cell which express hSCF and IL-15,the activity of supernantes was respectively assayed by TF-1 and CTLL-2 cell line. CD3, CD16, CD56, CD25, CD48, CD54, CD69 and CD95 molecules were tested by FACS.Results:We established NK-92S15 cell line which express hSCF and hIL-15 steadily, proliferation ability demonstrated it had more extensive assembling trend,growing with enough cell number,the cytotoxicity of NK-92S15 cell was decreased compared with parental cell line when incubated with rhIL-2.CD56 and CD16 showed on difference while CD25,CD48, CD54 and CD95 decreased significantly.Conclusion:The characterization of NK-92 could be changed by co-transfecting with hSCF and hIL-15 gene, which was benefit to observe the character of NK cell activation and cytotoxicity related molecules. The gene transfection of NK-92 cell made it suitable for further study.