1.Expression of recombinant human kallistatin in Pichia pastoris by high density cell culture, and its purification and characterization.
Qian ZHANG ; Yongmei XING ; Jia LIU ; Yong DIAO
Acta Pharmaceutica Sinica 2013;48(7):1107-12
Kallistatin (Kal) is a negative acute phase endogenous protein which can inhibit tumor angiogenesis, growth and metastasis effectively. To express and purify recombinant human kallistatin (rHKal), and characterize its biological activity, P. pastoris was transformed with pPIC9-Kal/GS115 (His4) to express rHKal. The fermentation was carried out in a 7.5 L bioreactor with high density cell culture. 1%-2% methanol was added to the medium to induce the expression of rHKal. The secretion was purified with phenyl sepharose, G-25 sepharose, heparin sepharose and Sephacryl S-100 chromatography. The biological activity of purified bulk rHKal on HUVEC was evaluated with MTT and tube formation assays. The final expression of rHKal in the supernatant reached 50 mg x L(-1), the purity of bulk rHKal after purification was above 98%. A dose-dependent inhibition of rHKal on HUVEC proliferation was observed, however, a U-shaped dose-response curve of rHKal on capillary formation of HUVEC was revealed. The described protocol provides an effective means for preparing rHKal that could be used for anti-angiogenesis therapy in the future.
2.Congenital cystic adenomatoid malformation of lung in adult
Xing-Mei ZHANG ; Kan LIU ; Shu-Lin JIA ; Gang CHENG ;
Chinese Journal of Radiology 2001;0(08):-
Objective To investigate of the radiological manifestations in congenital cystic adenomatoid malformation (CCAM) of lung in adult and to improve the diagnostic accuracy of CCAM of lung in adult.Methods Five cases with pathologically proved CCAM of lung in adult were retrospectively analyzed.Chest X-ray was available in 5 cases and chest CT was performed in 2 cases.Results On plain chest radiography,thin wall air cystic lesions with air-fluid level were detected in 3 cases.Honeycomb like small cystic lesion was detected in 1 case.Multiple round cystic lesions were found in 1 case.CT scan of the chest demonstrated a round thin wall air cystic lesion in the lateral segment of right lung's middle lobe,and a thin wall air cystic lesion with the wall merged into the shadow of chest wall in the left apicoposterior segment in one case.Small cystic lesions just like honeycomb were found in bilateral basal segments of the inferior lobes,with a 0.8—1.0 cm sized round mass revealed in the right lung′s inferior lobe basal segment, and the mass was spiculated in another case.Conclusion The imaging signs of CCAM of lung in adult is cyst or cyst-solid and at the risk of developing carcinoma.
3.Study of MTT and resazurin assays for susceptibility test to first-line anti-tuberculosis drugs
Zongde ZHANG ; Aiying XING ; Zhongquan LIU ; Hongyan JIA ; Shuxiang GU
Chinese Journal of Laboratory Medicine 2008;31(9):989-992
Objecflve To evaluate the performance of two rapid and low-cost metheds(MTT test,and rosazurin mierotitre assay)for the detection of resistance to first-line drugs in Mycobacterium tuberculosis.Methods sixty-four Myeobaeterium tuberculosis clinical isohtes were tested by the MTT test and the rosazuxin microtitre assay(REMA)respectively,and the results were compared with those obtained with the absolute concentration method on L(o)wenstein Jensen medium.Results The MTT test and the resazurin microtitre assay showed a good agreement compared with the absolute concentration method for all first-line drugs tested.The sensitibity,specificity and accuracy of the MTT test were 94.8%,96.0%,95.3%,for RFP;93.8%,93.8%,93.8% for INH;92.9%,96.O%,95.3% for EMB,90.6%,87.5%,89.1% for SM,respectively.The sensitivity,specificity and accuracy of the resazurin microtitre assay were 92.3%,96.0%,93.8%,for RFP;90.6%,90.6%,90.6% for INH;92.9%,94.0%,93.8% for EMB,87.5%,87.5%,87.5% for SM,respectively.The Kappa value of the MTT test and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.857,0.831,0.714,0.792.respeedvely;The Kappa value of the regazurin mierotitre assay and the absolute concentration method for the detection of resistance to RFP,INH,EMB,SM were 0.871,0.826,0.826,0.750,respectively.The Kappa value of the MTT test and the resazurin microtitre assay for the detection of resistance to RFP,INH,EMB,SM wefe 0.889,0.875.0.787,0.844,respectively.Conclusions Both MTT test and the resazurin microtitre assays are simple,rapid,low-cost and sensitive for rapid detection of resistance to first-line drugs.They could be promising methods for susceptibility assay of the first-line antituberculosis drugs in low-resource countries.
4.“T”-shaped locking plate for posterolateral tibial plateau fractures by Carlson posterolateral approach:12-month follow-up
Chong FENG ; Dailiang JIA ; Xuefeng LEI ; Gang ZHANG ; Qining XING
Chinese Journal of Tissue Engineering Research 2016;20(17):24471-24478
BACKGROUND:Single fracture or colapse of the posterolateral tibial plateau fractures is relatively rare in the clinical work. Rational choice of surgical approach and internal fixation for posterolateral plateau fracture is significant to restore the lower limb force line, maintain the joint stability and obtain good biocompatibility.
OBJECTIVE:To compare the stability and biocompatibility of Carlson posterolateral and posterior midline approaches for the treatment of posterolateral tibial plateau fractures with “T” shaped locking plate.
METHODS:From July 2011 to July 2014, 43 patients with posterolateral tibial plateau fractures, who were treated in the Affiliated Hospital of Jining Medical University, were retrospectively analyzed. Al patients were assigned to two groups according to approaches. In the Carlson posterolateral approach group, 22 cases received “T”-shaped plate insertion by Carlson posterolateral approach. In the posterior midline approach group, 21 cases received “T”-shaped plate insertion by posterior midline approach. After repair, perioperative data, fixation effects and knee function score were compared and analyzed between both groups.
RESULTS AND CONCLUSION:(1) 43 cases (43 knees) of posterolateral tibial plateau fractures were folowed up strictly. (2) No significant difference in operation time, fracture healing time, total load time, Hospital for Special Surgery score at 12 months postoperatively, tibial plateau angle and posterior slope angle immediately and 12 months postoperatively was detected between both groups (P > 0.05). (3) Significant differences in fracture exposure, blood loss, and excelent and good rate of Rasmussen at 12 months postoperatively were identified in both groups. Moreover, above indexes were better in the Carlson posterolateral approach group than in the posterior midline approach group (P< 0.05). (4) These findings confirmed that for a single fracture or colapse of the posterolateral tibial plateau fractures, two kinds of surgical approaches can achieve ful and direct exposure. Carlson posterolateral approach has good repair effect, fixation effect and biocompatibility.
5.Application of ultrasound-guided transversus abdominis plane block for operations of inguinal region in infants
Yongsheng QIU ; Defu ZHANG ; Yingping JIA ; Xing LI ; Qing XU
Chinese Journal of General Practitioners 2014;13(12):1016-1018
One hundred and fifty ASA Ⅰ-Ⅱ infants scheduled to undergo surgery on unilateral inguinal region,were randomly assigned to 3 groups with 50 in each group.In groups Ⅰ and Ⅱ puncture was performed at traversus abdominis plane under ultrasound-guidance,0.4 or 0.5/kg of 0.25% ropivacaine was injected respectively.In group Ⅲ the triangle of Petit was positioned through palpating and then 0.5 ml/kg of 0.25% ropivacaine was injected.HR,RR,SpO2,PETCO2,expired sevoflurane concentration and BIS value at the time of entering the operation room (T1),incision of skin (T2),pulling hernia sac (T3),ending the surgery (T4),waking (T5),the number of pressing the analgesia pump after operation were recorded.The results showed that HR at T2 and T3 of groups Ⅰ and Ⅱ was lower than group Ⅲ (P < 0.05).The were significant differences in case numbers of insufficient intraoperative analgesia among 3 groups(x2 =10.500,P =0.005).The CHEOPS scores and the number of pressing analgesia pump after operation in groups Ⅰ and Ⅱ were lower than those in group Ⅲ (x2 =7.230,P =0.027).Results indicate that ultrasound-guided transversus abdominis plane block is safer and more effective than conventional method for operations of inguinal region in infants; it may reduce dose of local anesthetics and postoperative use of analgesics.
6.Identification of Candidate Diagnostic Tumor Markers for Human Hepatocellular Carcinoma Using Genechip Technology
Xujian XING ; Huliang JIA ; Lei ZHANG ; Qinghai YE
Chinese Journal of Bases and Clinics in General Surgery 2004;0(01):-
Objective To identify genes associated with hepatocellular carcinoma (HCC) as candidate diagnostic markers in a genome-wide scale. Methods The gene expression profiles of 40 pairs of HCC tumor tissue and peripheral non-tumorous liver tissue were analyzed by using gene chip technology.The gene chips were fabricated at the National Cancer Institute (NCI). Each gene chip contained 9 180 genes. The fluorescent targets were prepared by a direct labeling approach using two kinds of fluorescences as following: 100 ?g of total RNA from non-cancerous liver tissue was labeled with Cy3-dUTP and 200 ?g of total RNA from HCC was labeled with Cy5-dUTP. The targets were mixed together and hybridized with genes on the gene chips. Unsupervised hierarchical clustering analysis was done by CLUSTER and TREEVIEW software using median centered correlation and complete linkage. Results A total of 10 genes were found up-regulated in over 80% of primary tumors comparing with that of their corresponding non-tumorous liver tissues at a two-fold filter with an unsupervised hierarchical clustering algorithm, including protocadherin-alpha 9, ESTs, Homo sapiens cDNA FLJ, KPNA2, RPS20, SNRPE, CDKN2A, UBD, MDK and ANXA2. Conclusion These genes are supposed to be candidates for the diagnosis of HCC. Further investigation of these genes in a large scale of patients with HCC and patients with non-malignant hepatic diseases will be needed to disclose whether they could be used clinically as novel diagnostic tumor markers for HCC.
7. Pantoprazole and ranitidine in treatment of duodenal ulcer: a systematic review and pharmacoeconomics analysis
Chinese Pharmaceutical Journal 2015;50(13):1160-1164
OBJECTIVE: To systematically review the effectiveness and safety of pantoprazole in comparison with ranitidine for patients with duodenal ulcer, and evaluate the cost of the two drugs. METHODS: PubMed, Medline, EMbase, the Cochrane Library and three Chinese literature databases (VIP, CNKI and WanFang) were searched. Randomized controlled trials (RCTs) that compared the clinical outcomes of pantoprazole (PAN) group vs. ranitidine (RAN) group for duodenal ulcer were included. Two reviewers independently screened literature in accordance with the inclusion and exclusion criteria, extracted the data and assessed the methodological quality of included studies. Then, meta-analysis was performed using RevMan 5.2 software and cost-effectiveness analysis was performed with the decision tree model. RESULTS: A total of six RCTs involving 1 225 patients were included. The results of meta-analysis showed that PAN was superior to RAN in the 2-week or 4-week ulcers healing rates [RR = 1.61, 95% CI (1.44, 1.80), P<0.000 01 and RR = 1.18, 95% CI(1.12, 1.23), P <0.000 01, respectively]. Compared with RAN groups, PAN group had significantly higher rates of pain and gastrointestinal symptoms relief[RR = 1.31, 95% CI (1.15, 1.39); P<0.000 01 and RR = 1.18, 95% CI(1.08, 1.28), P = 0.000 3, respectively]. While there was no significant difference in the incidence of adverse events between the PAN and the RAN groups [RR = 1.35, 95% CI (0.87, 2.10), P = 0.19]. And the cost-effectiveness ratio was lower in the RAN group than in the PAN group (29.33 vs 53.90). CONCLUSION: Current evidence indicates that, pantoprazole is an effective scheme for duodenal ulcer, and ranitidine is considered as an economic choice. Both treatments are safe and well tolerated.
8.Supplement of Dongyuan acupuncture-moxibustion therapy and clinical records.
Yunfang JIA ; Xianning HOU ; Xuaoping ZHANG ; Haijiao XING ; Xin ZHANG ; Yuzhu DU ; Chunsheng JIA
Chinese Acupuncture & Moxibustion 2016;36(2):212-216
Professor JIA Chunsheng believes that "Dongyuan's acupuncture therapy" proposed by GAO Wu could not entirely reflect LI Dongyuan's academic thought of acupuncture and moxibustion. Hence, we collected all of the academic works of LI Dongyuan and extracted the texts relevant with acupuncture-moxibustion therapy, titled as Dongyuan's acupuncture-moxibustion therapy. This process contributed to the deeper study on Dongyuan's acupuncture and moxibustion academic thought. By the influence of Dongyuan's acupuncture-moxibustion academic thought, in association with his clinical experience for dozens of years, Professor JIA Chunsheng proposed that acupuncture and moxibustion should pay attention to the regulation and tonification of spleen and kidney. In clinical practice, Zhongwan (CV 12), bilateral Tianshu (ST 25), Qihai (CV 6) and Guanyuan (CV 4) are commonly selected to consolidate the root, cultivate the primary and treat the spleen and kidney simultaneously. The locations of the above selected points are distributed like a star. This "star-like point selection method" points to the importance of the spleen and kidney (the primary qi). For many diseases, especially the disorders of internal injury, the star-like acupuncture therapy is commonly used and the relevant acupoints are supplemented. The good clinical efficacy has been achieved.
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9.Study on the association of serum matrix metalloproteinase-9 and white blood cell count to type 2 diabetes mellitus with macroangiopathy and the interventing treatment
Jia LI ; Lv-Zhi JIANG ; Xing-Jia ZHOU ; Xue-Ning ZHANG ; Zheng-Fang LI ;
Chinese Journal of Endocrinology and Metabolism 2001;0(05):-
Objective To evaluate the relationship between serum matrix metalloproteinase-9(MMP-9), white blood cell(WBC)count and type 2 diabetes mellitus with macroangiopathy and to investigate the mechanism of the protective effects of rusiglitazone(RSG)on blood vessel.Methods Serum MMP-9 was determined by ELISA in 30 normal controls and 80 type 2 diabetic patients(including 40 cases with macroangiopathy and 40 without maeroangiopathy).WBC count and other clinical parameters were also determined.32 type 2 diabetic patients received RSG(4rag qd)for 12 weeks.All parameters were determined after 4 weeks(17 patients)and after 12 weeks(all patients)to observe the changes in MMP-9,WBC and other parameters.Results In the diabetic patients,serum MMP-9 and WBC were markedly higher as compared with normal controls;and MMP-9 and WBC in patients with macroangiopathy[579(440-949)?g/L,(7.51?1.47)?10~9/L]were higher than those [324(275-423)?g/L,(6.22?0.79)?10~9/L]in the cases without macroangiopathy(P
10.Initial experience of transumbilical laparoendoscopic single-site surgery in urology
Xing AI ; Zhuomin JIA ; Feng GAO ; Shuiwen ZHANG ; Fengling SUN ; Guohui ZHANG ; Tong ZANG ; Xu ZHANG
Chinese Journal of Urology 2012;33(2):96-98
ObjectiveTo summarize the initial experience of transumbilical laparoendoscopic single-site surgery of urology.MethodsFrom February 2010 to March 2011,21 patients underwent laparoendoscopic single-site surgery using transumbilical single-site and common surgical instruments of laparoendoscopic.Nine patients underwent single-site laparoscopic ureterolithotomy,5 underwent transumbilical single-site laparoscopic ureteral stricture resection and anastomosis,5 underwent transumbilical single-site laparoscopic renalcyst unroofing and 2 had a nephrectomy.All of the cases were definitely diagnosed.A single umbilical incision of 1.5 cm to 2.5 cm was made for Triport.The procedures were performed according to the methods used in classical laparoscope methods using general instruments.ResultsAll the operations were successfully completed without conversion to open surgery.The mean operative time of ureterolithotomy was 143 (120-230) min,the mean operative time of ureteral stricture resection and anastomosis was 157 (120 -180) min,the mean operative time of unroofing of renal cysts was 110 (95 -132) min,and the operative time of the nephrectomy was from 95 to 120 min.The intestinal tract function recovered within 1 -2 d,the drainage tube was removed within 2 -3 d and the postoperative hospitalization duration was 4 -7 d.The symptoms were reduced or disappeared and no major intraoperative or postoperative complications occurred within 4 - 6 months.Conclusions Transumbilical laparoendoscopic single-site surgery represents a safe and feasible operation for urologic patients.With more clinical practice,laparoendoscopic single-site surgery could be generally applied.