1.Construction and identification of recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 antigen
Xuemei WANG ; Jiangkun LUO ; Qian LI ; Jiangyan LI ; Yong CHEN ; Zhiyong TAO ; Hui XIA ; Qiang FANG
Chinese Journal of Schistosomiasis Control 2014;(3):287-291
Objective To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 anti-gen. Methods The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction en-zymes,and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both frag-ments were modified by Klenow fragment to form blunt end,then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1-Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction,a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band,which sug-gested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis,the recombi-nant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombi-nant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.
2.Effect of curcumin on apoptosis in spinal cord and dorsal root ganglion neurons in a rat model of diabetic neuropathic pain
Guo CHEN ; Congcong HUANG ; Jiangkun DANG ; Qingquan LIAN ; Jun LI ; Hong CAO
Chinese Journal of Anesthesiology 2011;31(4):435-438
Objective To investigate the effect of curcumin on the apoptosis in spinal cord and dorsal root ganglion neurons in a rat model of diabetic neuropathic pain (DNP) . Methods One hundred and eight male SD rats weighing 200-230 g were randomly divided into 4 groups ( n = 27 each): control group (group C), DNP group, solvent control group (group SC) and curcumin group (group Cur) . Diabetes was induced with intraperitoneal streptozocin 70 mg/kg. Successful induction of diabetes was defined as blood glucose > 16.7 mmol/L. Curcumin and com oil 100 mg/kg (23 mg/ml) were given intraperitoneally once a day for 14 consecutive days starting from 14 days after administration of streptozocin in Cur and SC groups respectively. Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 2 d before and 14 d after streptozocin injection and 3, 7 and 14 d after curcumin injection. The pain threshold measured at 14 d after administration of streptozocin decreased by more than 15% of the baseline in all the rats. The expression of caspase-3 and Bcl-2 in spinal cord and dorsal root ganglion was determined at 3, 7 and 14 d after curcumin injection by immuno-histochemistry and Western blot, and the neuronal apoptosis rate was determined by TUNEL. Results Compared with group C, MWT and Bcl-2 expression were significantly decreased, TWL was significantly shortened, the neurona lapoptosis rate and caspase-3 expression were significantly increased in DNP, SC and Cur groups ( P < 0.05).Compared with group DNP, MWT and Bcl-2 expression were significantly increased, TWL was significantly prolonged, the neuronal apoptosis rate and caspase-3 expression were significantly decreased in Cur group ( P <0.05) . There was no significant difference in the parameters mentioned above between DNP and SC groups ( P >0.05). Conclusion Curcumin can attenuate DNP by inhibiting the apoptosis in spinal dorsal hom and dorsal root ganglion neurons in rats, and the inhibition of caspase-3 expression and increase in Bcl-2 expression are involved in the mechanism.
3.Effects of curcumin on type 2 diabetic neuropathic pain and expression of inositol-requiring enzyme 1α in spinal dorsal horn and dorsal root ganglia in rats
Jiangkun DANG ; Xiaodan WANG ; Lin ZHOU ; Hong CAO ; Yan WU ; Jun LI ; Qingquan LIAN
Chinese Journal of Anesthesiology 2012;32(6):676-679
ObjectiveTo investigate the effects of curcumin on type 2 diabetic neuropathic pain (DNP)and expression of inositol-requiring enzyme 1α (IRE1α) in spinal dorsal horn and dorsal root ganglia (DRG) in rats.MethodsType 2 diabetes mellitus was induced by high-fat and high-sucrose diet and intraperitoneal streptozotocin (STZ) 35 mg/kg,and confirmed by fasting blood glucose level > 16.7 mmol/L in male SD rats.Type 2 DNP was confirmed by the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWI.) measured on day 14 after STZ administration < 80% of the baseline value.The rats were then randomly divided into 3 groups ( n =27 each):DNP group,curcumin group (group Cur) and solvent control group (group SC).Curcumin and corn oil 100 mg/kg (25 mg/ml) were given intraperitoneally once a day for 14 consecutive days starting from 14 days after administration of streptozocin in Cur and SC groups respectively.Another 27 rats were chosen and served as control group (group C) and fed with common fodders.MWT and TWL were measured at 3,7 and 14 day after curcumin administration.The expression of IRE1α in spinal dorsal horn and DRG was detected by Western blot.ResultsCompared with group C,MWT was significantly decreased and TWL was significantly shortened,and the expression of IRE1α was up-regulated in DNP,Cur,and SC groups (P < 0,05),Compared with group DNP,MWT were significantly increased,TWL was significantly prolonged,and the expression of IRE1α in spinal dorsal horn and DRG was down-regulated in group Cur (P < 0.05).There was no significantdifference in the parameters mentioned above between DNP and SC groups (P > 0.05).ConclusionCurcumin can attenuate type 2 1)NP and inhibition of the expression of IRE1α in spinal dorsal horn and DRG is involved in the mechanism.
4.Associating liver partition and portal vein ligation for staged hepatectomy combined with hepatic artery reconstruction in the treatment of hilar cholangiocarcinoma
Yaoxuan WANG ; Ke LI ; Yuwei TIAN ; Qingfeng JIANG ; Quan SHEN ; Jiangkun JIA ; Huanzhou XUE
Chinese Journal of Hepatobiliary Surgery 2018;24(9):600-603
Objective To study the combined use of ALPPS (associating liver partition and portal vein ligation for staged hepatectomy) with hepatic artery reconstruction in the treatment of hilar cholangiocarcinoma with hepatic arterial involvement.Methods The clinical data of 7 patients with hilar cholangiocarcinoma who underwent ALPPS combined with hepatic arterial resection and reconstruction were analyzed retrospectively.The technical points and the perioperative management were analyzed.Methods At the first stage,the relationship between the tumor and the vessels were explored,the portal vein of the part of the liver to be resected was ligated and the liver was transected with a CUSA (Cavitron Ultrasound Surgical Aspirator).Then the bile duct was cut and a hepaticojejunostomy was completed.Finally,under ultrasound guidance,a bile duct drainage tube was inserted transhepatically into the part of the liver which was to be resected.Two to three weeks later,and after enough hypertrophy of the liver remnant size was confirmed,tumor resection was completed with reconstruction of the hepatic artery.Results Seven patients underwent the second stage operation,with no perioperative death.Six patients developed pulmonary infection and were treated successfully with conservative treatment.Two patients developed postoperative bile leak with secondary abdominal infection.One patient developed postoperative hepatic artery thrombosis secondary to biliary tract infection.Conclusion ALPPS combined with hepatic artery reconstruction was safe and feasible in the treatment of hilar cholangiocarcinoma with hepatic arterial involvement.
5.Development and performance test of unmanned aerial vehicle-borne CeBr3 radiation monitoring system
Jiangqi FANG ; Zhengwei AN ; Wenfeng ZHANG ; Linfeng LIU ; Jinzheng YANG ; Xue LIU ; Jiangkun LI
Chinese Journal of Radiological Health 2024;33(1):80-86
Objective To develop an unmanned aerial vehicle (UAV)-borne radiation monitoring system with high detection efficiency and nuclide identification ability for airborne monitoring in nuclear emergency. Methods The UAV-borne CeBr3 radiation monitoring system was composed of four cerium bromide (CeBr3) crystal detectors coupled with silicon photomultipliers (SiPMs) and other components including integrated modules, intelligent electronic devices, and new composite materials. Results According to various performance tests on the system, the crystal energy resolution was better than 5% (@0.662 MeV), the peak drift of the energy spectrum was within ±1 channel, the linear fit of energy was 0.99997, the change in the count rate of each energy window during 12 h long-term measurement was less than 5%, and the detection efficiency was higher compared with that of NaI (Tl) detectors of the same volume. Conclusion Through ground point source testing and theoretical calculation, the system has reliable ability to identify radionuclides, which can be used in nuclide identification and the preparedness and response for nuclear and radiation emergencies.