1.Application of MTT Colorimetric Assay in Research of Cytokine Activity and Cytotoxicity of Killer Cells
Chinese Journal of Cancer Biotherapy 1994;0(01):-
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.
2.Clinical Research of Changes of Extrahepatic Bile Duct Diameter Before and after Laparoscopic Cholecystectomy
Hao CHEN ; Jianwei TIAN ; Jianhua LI
Chinese Journal of Bases and Clinics in General Surgery 2008;0(10):-
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.
3.Recent advance of the interventional treatment of refractory ascites
Wen LI ; Xuelong TIAN ; Jianhua WANG
Chinese Medical Equipment Journal 1989;0(02):-
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.
4.Novel gene sequence variants of pbp2b in penicillin-nonsusceptible Streptococcus pneumonia isolates
Sufei TIAN ; Yunzhuo CHU ; Baiyi CHEN ; Jianhua WAN
Chinese Journal of Internal Medicine 2008;47(6):491-494
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.
5.Correlation between T help lymphocyte and autonomic nervous system in patients with congestive heart failure
Xiuqing TIAN ; Jiangjiu LIANG ; Jianhua LIU ; Yuling DONG ; Bingyu WU
Clinical Medicine of China 2010;26(10):1023-1025
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.
6.Dynamic Enhanced CT Diagnosis of Hilar Cholangiocarcinoma
Tian WU ; Jianhua YAN ; Zhanguo XU ; Xiaofei QU
Journal of Practical Radiology 1992;0(11):-
Objective To assess the value of spiral CT in the diagnosis of hilar cholangiocarcinoma.Methods 13 patients of hilar cholangiocarcinoma underwent CT plain and dynamic enhanced scan.CT findings were analysed in comparison with that of surgery and pathology.Results On plain scan,the lesions appeared as soft tissue mass with low density in hepatic hilar(n=11),bile duct wall irregularly thickened and bile duct narrowed(n=2).On dynamic enhanced scan,delayed enhancement was seen in 13 cases.Conclusion Plain spiral CT and dynamic enhanced scan have great value in showing the morphological features and improving the diagnosis of hilar cholangiocarcinoma.
7.The characterization of NK cell line co-transfected with human stem cell factor and human interleukin-15 gene
Jinbao ZONG ; Jian ZHANG ; Jianhua ZHANG ; Rui SUN ; Zhigang TIAN
Chinese Journal of Immunology 2000;0(09):-
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.
8.Radical nephrectomy with resection of involved adjacent organs (report of 24 cases)
Ming LI ; Shengtao HONG ; Jianhua TIAN ; Dong LU
Chinese Journal of Urology 2001;0(03):-
Objective To evaluate the feasibility and clinical significance of radical nephrectomy with en bloc resection of involved adjacent organs.Methods Totally,24 cases of renal cancer invading adjacent structures underwent radical nephrectomy with en bloc resection of involved adjacent organs.Among the 24 cases,left radical nephrectomy with en bloc resection of splenic flexure and descending colon was performed in 7,partial resection of corpus and cauda pancreatis and spleen in 5,solitary splenectomy in 3;right radical nephrectomy with en bloc resection of hepatic flexure of the colon in 4,partial hepatectomy of right lobe and end-piece in 4,duodenectomy of pars descendens in 1.Of these cases,partial resection of the psoas muscle was performed in 5,and partial resection of mesocolon in 7.Postoperatively,9 cases received immunotherapy.Results There was no intraoperative mortality or severe posuoperative complication.Follow-up of 3-240 months was available in 21 cases.The follow-up showed that 1-,3-,5-and 8-year survival rates were 90.5%(19/21),42.9%(9/21),33.3%(7/21) and 19.0%(4/21),respectively.Conclusions Radical nephrectomy remains the treatment of choice in organ-confined stage of renal carcinoma.With careful selection,radical nephrectomy with en bloc resection of adjacent structures is technically feasible.It can obtain the radical excisional effect.Based on our experience,the operation is relatively safe.Complete surgical extirpation can lead to prolonged disease-free survival.It may also offer beneficial foundation for the subsequent systematic therapy.
9.Construction and screening of human AQP1 shRNA expression vectors
Zhuo LI ; Wei KANG ; Na XIN ; Yu TIAN ; Jianhua LI
Chongqing Medicine 2015;(30):4183-4186
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 .
10.Combination of multi-disciplinary techniques with ~(125)Ⅰ seeds in treating malignant obstructive jaundice
Xueming DU ; Jianhui XU ; Jianhua LANG ; Xiurong TIAN ; Wei DONG
Journal of Interventional Radiology 2001;0(05):-
0.05).Conclusion Multi-disciplinary techniques combined with 125Ⅰ seeds implantation is effective in the management of the malignant obstructive jaundice.No significant difference for relief and liver function were found between CT-guided and during operation interstitial 125Ⅰ seeds implantations,but it seems more quickly relief or recovery was achieved in the latter.