1.Construction and expression of mouse neuromedin B and neuromedin B receptor gene overexpression vectors
Chinese Journal of Biologicals 2024;37(11):1294-1299
Objective To construct the lentiviral overexpression vectors of mouse neuromedin B(NMB) and neuromedin B receptor(NMBR) genes and express them in RAW264.7 cells, so as to lay a foundation for further study on the effects of mouse NMB and NMBR gene overexpression on the proliferation and apoptosis of RAW264.7 cells. Methods The coding sequences(CDSs) of mouse NMB and NMBR genes were amplified by RT-PCR and inserted into vector pCD513B-1 to construct recombinant plasmids pCD513B-1-NMB and pCD513B-1-NMBR. Mouse NMB and NMBR gene overexpression lentiviruses were obtained through packaging by HEK-293T cells, and the virus titers were detected by double dilution method. After infection with lentivirus for 48 h, RAW264.7 cells were detected for the expression of NMB and NMBR mRNA by qPCR using the uninfected cells as control. Results The recombinant plasmids were constructed correctly as identified by double enzyme digestion. The virus titers of mouse NMB and NMBR gene overexpression lentiviruses were 6 × 106and 8 ×106TU/mL, respectively. The mRNA expression levels of mouse NMB and NMBR genes in RAW264.7 cells transfected with two lentiviruses were significantly higher than those in the control group(t = 24. 158 and 14. 958, respectively, each P <0. 01). Conclusion Mouse NMB and NMBR gene overexpression vectors were successfully constructed, which can significantly increase the expression of NMB and NMBR genes in RAW264.7 cells.
2.Diagnosis and treatment of renal cell carcinoma complicated with inferior vena cava tumor thrombus in hepatic segment and upper hepatic segment
Chinese Journal of Urology 2017;38(7):481-484
Surgical treatment is the only cure treatment for patients with inferior vena cava tumor thrombus in hepatic segment and upper hepatic segment.The accurate diagnosis of tumor thrombus is very important.In preoperative imaging examination,the abdominal enhanced CT scan and the inferior vena cava MRI scan were the best methods for the diagnosis and evaluation of the tumor thrombus in hepatic segment and upper hepatic segment.Compared with the tumor thrombus below the liver,the tumor thrombus in hepatic segment or above hepatic segment extend widely,and the operation are more difficult.For simple inferior vena cava tumor thrombus (the top of the thrombus has reached the level of hepatic vein),Retroperitoneal approach combined with transperitoneal approach should be used.Open surgery is the standard procedure for other tumor thrombus in hepatic segment and upper hepatic segment.In addition to exposure of inferior vena cava below the hepatic vein,the liver and the first hepatic hilum should be exposed.For tumor thrombus in the atrium,after the longitudinal incision of diaphragm,we use Milking technology to squeeze thrombus into inferior vena cava.Then we use catheterization technology to remove thrombus.For difficult atrial tumor thrombus,an extracorporeal circulation should be performed.The median incision in the chest should be performed to open the chest and open the pericardium and remove the tumor thrombus.Patients with tumor thrombus in hepatic segment or upper hepatic segment should be diagnosed as early as possible and they need actively treated by operation.
3. Protection of 20-hydroxyecdysone on SH-SY5Y cells from 1-methy-4-phenylpyridinium-induced injury
Chinese Pharmaceutical Journal 2013;48(22):1923-1927
OBJECTIVE: To investigate the protective effect and probable mechanism of 20-Hydroxyecdysone on SH-SY5Y cells injured by MPP+. METHODS: MPP+ induced SH-SY5Y cells injury model was established. 20-Hydroxyecdysone was added into the culture to test its protective effect. The morphological changes of cells were observed. Cell viability was detected by method of MTT. The contents of LDH, MDA and activity of SOD were inspected by kits. Apoptosis rate of cells was detected by flow cytometry. RESULTS: Compared with MPP+ group, 20-Hydroxyecdysone group (50, 100, 150 μmol·L-1) alleviated the damage of SH-SY5Y cells induced by MPP+, significantly improved cell survival rate, reduced the release of LDH, increased the activity of SOD, decreased the contents of MDA and reduced the apoptosis of cells. CONCLUSION: 20-Hydroxyecdysone has a significant protective effect on SH-SY5Y cells injured by MPP+. The probable mechanism may be anti-oxidation and anti-apoptosis.
4.Effect of ABCCA gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3
Jin MA ; Zhuo ZHANG ; Jiancheng WANG
Chinese Journal of Pancreatology 2013;13(4):252-255
Objective To determine the effect of ABCC4 gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3.Methods PANC1 and BxPC-3 pancreatic cancer cells were transfected with a lentivirus expressing an ABCCA short hairpin RNA (shRNA).ABCC4 mRNA and protein expression of transfected cells was determined by RT-PCR and Western blot,colony formation ability was measured by colony assay,and cell cycle change was investigated by the flow cytometric analysis.Results Lentivirus expressing an ABCC4 short hairpin RNA (shRNA) was successfully established.After transfection with shRNA lentivirus,ABCC4 mRNA and protein expression were significantly inhibited (0.28 ±0.01 vs 1.00 ±0.03,0.22 ±0.02 vs 1.00 ±0.03,P <0.05).Colony formation ability was significantly decreased (4 vs 65,P <0.05),and cell cycle was significantly blocked at G1 phase [(54.98 ±1.78) % vs (42.93 ± 0.88) %,(68.55 ± 0.75) % vs (54.76 ± 0.29) %].Conclusions ABCC4 gene silencing can significantly inhibit cell proliferation of human pancreatic cancer cell lines PANC1 and BxPC-3,and block the cells at G1 phase.
5.The Level and Significance of Serum IL-18 and sFas/sFasL in Patients with COPD
Songmin ZHUO ; Ruihuan XU ; Zhen MA
Journal of Chinese Physician 2001;0(09):-
Objective To investigate the level change and correlation of serum IL-18 and soluble Fas/Fas ligand(sFas/sFasL) in patients with chronic obstructive pulmonary disease (COPD) to evaluate their roles in the pathogenesis of COPD. Methods The levels of serum IL-18 and sFas/sFasL in 36 patients with COPD of acute aggravation stage and 20 healthy control subjects were detected by ELISA. Results The levels of serum sFasL and IL-18 in patients with COPD were significantly lower than those in healthy control subjects, while there was not significant differene in serum sFas level between the two groups. Conclusion The serum sFasL and IL-18 level decreass in patients with COPD of acute aggravation stage indicated that cell apoptosis level and immune function reduced in the patients.
7.Estimation of the entrance surface dose from the dose indicator in computed radiography system
Tianjun MA ; Bo CHEN ; Weihai ZHUO ; Haikuan LIU
Chinese Journal of Radiological Medicine and Protection 2013;33(4):423-426
Objective To study the relationship between the dose indicator of computed radiography (CR) and the entrance surface dose (ESD),and to build a model for estimating ESD based on this relationship.Methods Taking Kodak CR system as the research object,a theoretical model for estimating the ESD was established according to theoretical derivations,and the key parameters in the model were determined through experiments in a CR system.Further experiments in another CR system were also conducted to verify the model.Results The ESDs were not only dependent on the dose indicator provided by the CR system,but also influenced by other factors,such as tube potential,patient's body thickness and energy response of the imaging plate.The estimation results of the model agreed well with the experiment results,and the relative deviation was confirmed within 20%.Conclusions The dose indicator based model can provide a relatively fast and easy way for evaluating the doses of patients undergoing X-ray diagnoses with the CR system.
8.Efficacy of choledochoscopy for the treatment of postoperative intrahepatic duct stones and strictures
Yuehua GUO ; Zhuo ZHANG ; Yang MA ; Shiyun BAO ; Xiaofang YU
Chinese Journal of Digestive Surgery 2013;12(8):616-619
Objective To summarize the experience of surgical management of postoperative intrahepatic duct stones and strictures with choledochoscope.Methods The clinical data of 1241 patients with intrahepatic duct stones and strictures who were admitted to the Shenzhen People's Hospital from January 2000 to December 2011 were retrospectively analyzed.All patients were divided into postoperative choledochoscopy group (652 patients were treated by choledochoscope through T-tube fistula) and conservative treatment group (589 patients were treated by cholangic and antibiotic drugs) according to treatment plan.The clinical data including stricture alleviation,incidence of complications and treatment efficacies of the 2 groups were compared using the chi-square test.Results In the postoperative choledochoscopy group,the intrahepatic duct stones of 630 patients were completely removed,with the removal rate of 96.63% (630/652),which was significandy higher than 76.23% (449/589) of the conservative treatment group (x2=113.407,P < 0.05).A total of 1237 intrahepatic duct strictures were detected in the postoperative choledochoscopy group,including 698 cases of membranous stricture (the length of strictured bile duct < 2 mm),529 cases of spool-shaped stricture (2 mm ≤ the length of strictured bile duct ≤5 mm),8 cases of long tube-shaped stricture (the length of strictured bile duct > 5 mm),2 cases of crackshaped stricture.The alleviation rate of the stricture was 99.35% (1229/1237).The conditions of 6 cases of long tube-shaped stricture and 2 cases of crack-shaped stricture were not alleviated.There were 986 cases of intrahepatic duct strictures in the conservative treatment group,and the stricture alleviation rate was 80.43% (793/986).There was a significant difference in the stricture alleviation rate between the 2 groups (x2=238.994,P < 0.05).Nine patients in the postoperative choledochoscopy group had severe complications.Five patients had intrahepatic duct hemorrhage with a volume of blood loss more than 1500 ml,and they were treated by balloon dilatation hemostasis.One patient with repeated intrahepatic duct bleeding received hepatectomy.Two patients with severe intrahepatic cholangitis received choledochoscopy.Two patients had incomplete formation of sinus and they received implantation of biliary stents under duodenoscope.Conclusion Standard surgical management with choledochoscope can effectively alleviate postoperative intrahepatic duct stones and strictures.
9.Phenotypic modulation of vascular smooth muscle cells in diabetes mellitus and intervention of traditional Chinese medicines.
Xiao-dong MA ; Zhuo-an CHEN ; Jian-pu ZHENG
China Journal of Chinese Materia Medica 2014;39(24):4723-4727
Proliferation and migration of vascular smooth muscle cells (VSMC) are common pathological features of diabetic vascular complications,such as atherosclerosis and hypertension. Phenotypic modulation of VSMC is the basis for VSMC proliferation and migration. Therefore, studies on VSMC phenotypic modulation and its mechanisms in diabetes mellitus were of important significance to the prevention and therapy of diabetic vascular complications. This paper introduces VSMC phenotypic modulation and the underlying mechanisms in diabetes mellitus, and summarizes advance of studies on traditional Chinese medicine intervention upon VSMC phenotypic modulation, so as to provide reference for preventing and treating diabetic vascular complications with traditional Chinese medicines.
Atherosclerosis
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drug therapy
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prevention & control
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Cell Movement
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drug effects
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Cell Proliferation
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drug effects
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Diabetes Mellitus
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drug therapy
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pathology
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Drugs, Chinese Herbal
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therapeutic use
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Humans
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Hypertension
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drug therapy
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prevention & control
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Medicine, Chinese Traditional
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Muscle, Smooth, Vascular
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drug effects
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Myocytes, Smooth Muscle
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drug effects
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Phenotype
10.Dietary intake level of elements with Kashin-Beck disease areas in parts of Aba Autonomous Prefecture of Sichuan Province
Ling LIU ; Xin ZHAO ; Wenjun MA ; Zize ZHUO
Chinese Journal of Endemiology 2014;(4):392-396
Objective To investigate and determine the dietary intake level and food source of elements in Kashin-Beck disease(KBD) and non-endemic KBD areas in parts of Aba Autonomous Prefecture , Sichuan Province. Methods Three KBD villages were selected from each county including the counties of Songpan , Jiuzhaigou and Ruoergai in Aba Autonomous Prefecture with multi stage random sampling method in 2010 , and 122 KBD patients who were diagnosed at county-level or higher-level hospitals were selected from villages. One non-KBD village was selected randomly from each county and 43 controls without KBD or other bone and joint disease were selected in proportion to the number of investigated KBD patients. Dietary intake of each object was recorded 24 hours before the survey using a 24 hour dietary survey method and intake of each kind of food(cooked food) was recorded. Three day dietary intake of each surveyed object was recorded in continuous and the average value was calculated. The local resident representative food samples were collected and 20 kinds of elements in these samples were determined with inductively coupled plasma mass spectrometry ( ICP-MS ) and inductively coupled plasma atomic emission spectrometry(ICP-AES), and daily elements intake was calculated and compared with the relevant intake standards. Results Daily intakes of vegetables and meat were 126.72, 28.25 g, respectively, in patients with KBD. More than 3 times of brick tea was drank by KBD patients than non-KBD people (P<0.05). Per capita daily intakes of Ca, Fe, Mg, K, Na, Zn, I, Cu and Co were less than recommended nutrient intake or allowable daily intake in both groups. More than 80% patients with KBD overtook more Al and Pb than tolerable upper intake levels. Conclusion There is a single dietary structure and a serious shortage of a variety of essential elements intake in parts of Aba Autonomous Prefecture, and the dietary intake level of elements is different between two groups.