1.Expression of DNA methyltransferase 1 mRNA in patients of endemic arsenism and its clinical significances
Xue-li, PAN ; Ai-hua, ZHANG ; Xiao-xin, HUANG
Chinese Journal of Endemiology 2010;29(1):13-17
Objective To investigate the transcription and expression of DNA methyltransferase 1 (DNMT1) mRNA in endemic arsenism patients by burning coal usage,to probe its effects on the development and carcinogenesis of arsenism. Methods In 2008,68 arsenism patients(including 24 mild cases,28 moderate cases and 16 severe cases) were selected in the areas with endemic arsenism according to Standarding of Diagnosis for Endemic Arsenism from Xingren county,Guizhou province. Among the subjects,40 cases were diagnosed by pathological methods,and they were divided into general pathological changes(20),precancerous(14) and cancerous group(6). Tweleve kilometer away from the endemic arsenism area,23 controls were selected in Daguoduo village (non-arsenism exposure). Under the principle of informed consent,blood samples were collected from individuals. The mRNA expression of DNMTI was detected by real-time quantitative reverse transcription polymerase chain reaction(FQ-PCR). At the same time,skin tissue samples were collected from the voluntary surgical treatment patients with endemic arsenism (total 61 cases,including 34 general pathological changes cases,21 precancerous cases and 6 cancerous cases) and from the control(15 cases). DNMT1 protein was detected by immunohistochemical method.Results Average level of DNMT1 mRNA were 0.221 83±0.595 09,0.246 11±0.509 79 and 0.389 27±0.411 33 respectively among mild,moderate and severe arsenism group. DNMT1 mRNA level of mild and moderate group were obviously lower than the control group(0.695 95±0.463 98,all P < 0.01). The mRNA average level of DNMT1 were 0.320 64±0.547 46,0.313 09±0.529 13 and 0.159 07±0.342 56 individually among general pathological changes,precancerous and cancerous group,which were obviously lower than the control group(0.695 95±0.463 98,all P < 0.05). The expression rates of DNMT1 protein in skin were 88.24%(30/34),100%(21/21) and 100% (6/6) among general pathological changes,precancerous and cancerous group were higher than the control group [0(0/15),all P < 0.01],and the extent of expression gradually increased with the aggravation of skin damage(r,= 0.740,P < 0.01). Conclusions DNMT1 participated in the development of the arsenism. High expression of its protein was an early event during the process of the arsenism. DNMT1 may be the new target markers for early diagnosis and treatment of arsenism.
2.Simultaneous determination of amlodipine, benazepril and benazeprilat in human plasma by LC-HESI/MS/MS method.
Hua-Ling PAN ; Li-Shan LIN ; Jue-Fang DING ; Xiao-Yan CHEN ; Da-Fang ZHONG
Acta Pharmaceutica Sinica 2014;49(1):95-100
The study aims to develop a rapid, sensitive and specified method of liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) for simultaneous determination of amlodipine, benazepril and benazeprilat in human plasma using amlodipine-d4 and ubenimex as internal standards (ISs). Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the positive mode for mass spectrometric detection. Analytes and ISs were extracted from plasma by simple protein precipitation. The reconstituted samples were chromatographed on a C18 (100 mm x 4.6 mm, 5 microm) column with mixture of methanol-acetonitrile-5 mmol.L- ammonium acetate-formic acid (30 : 30 : 40 : 0.1) as mobile phase at a flow rate of 0.6 mL.min-1. The standard curves were demonstrated to be linear in the range of 0.02 to 6.00 ng.mL-1 for amlodipine, 0.2 to 1,500 ng.mL-1 for benazepril and benazeprilat with r2>0.99 for each analyte. The lower limit of quantitation was identifiable and reproducible at 0.02, 0.2 and 0.2 ng mL-1 for amlodipine, benazepril and benazeprilat, respectively. The intra-day and inter-day precision and accuracy results were within the acceptable limit across all concentrations. The plasma samples were stable after four freeze-thaw cycles and being stored for 93 days at -20 degrees C. The method was applied to a pharmacokinetic study of a fixed-dose combination of amlodipine and benazepril on Chinese healthy volunteers.
Administration, Oral
;
Amlodipine
;
administration & dosage
;
blood
;
Benzazepines
;
administration & dosage
;
blood
;
Chromatography, Liquid
;
Humans
;
Sensitivity and Specificity
;
Spectrometry, Mass, Electrospray Ionization
;
Tandem Mass Spectrometry
3.Efficacy observation of combination of Lumbrukinase Capsule and Probucol Tablet in treating cerebral infarction patients' unstable atheromatous plaque of the carotid artery.
Xiao-feng PAN ; Wei SHAO ; Guo-hua CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(11):1479-1482
OBJECTIVETo observe the effects of combination of Lumbrukinase Capsule (LC) and Probucol Tablet (PT) in treating cerebral infarction (CI) patients' unstable atheromatous plaque of the carotid artery.
METHODS150 patients were randomly assigned to the PT group and the LC group, 75 cases in each. Patients in the PT group took 0.5 g PT each time, twice daily. On the basis of PT, patients in the LC group also took 600 thousand U LC, thrice daily. The treatment course was 12 months for all. The serum levels of TC, TG, HDL-C, LDL-C, fibrinogen (FIB), and changes of the carotid atherosclerotic plaque were measured before treatment, 6 and 12 months after treatment. Meanwhile, adverse events were recorded.
RESULTSThe serum levels of TC, TG, and LDL-C were all lower 6 months after treatment than before treatment in the two groups, showing statistical significance (P < 0.05). The serum level of HDL-C was higher 6 months after treatment than before treatment in the two groups, showing no statistical significance (P > 0.05). When compared with before treatment in the same group, the serum level of FIB significantly decreased after treatment. Besides, there was statistical difference between the two groups (P < 0.05). There was no statistical difference in the serum levels of blood lipids or FIB between 12-month treatment and 6-month treatment in the same group (P > 0.05). The plaque effective rate in the LC group was superior to that of the PT group, showing statistical significance (P < 0.01). During the treatment period, the occurrence of cerebrovascular event was lower in the LC group than in the PT group (P < 0.05). Partial patients in the two groups had gastric discomfort.
CONCLUSIONSThe combination of LC and PT could prevent and treat arteriosclerosis, stabilize the plaque, effectively lower the occurrence of ischemic events. Its clinical application did not increase the risk of hemorrhage. It was safe and effective, worthy of spreading. It was necessary to further observe whether combination of LC and PT could increase side effects of the digestive tract.
Adult ; Aged ; Animals ; Biological Products ; therapeutic use ; Carotid Artery Diseases ; drug therapy ; Cerebral Infarction ; drug therapy ; pathology ; Female ; Humans ; Male ; Materia Medica ; therapeutic use ; Middle Aged ; Oligochaeta ; enzymology ; Plaque, Atherosclerotic ; drug therapy ; Probucol ; therapeutic use ; Serine Endopeptidases ; therapeutic use ; Treatment Outcome
4.Clinical applications of distal artery island fascial flap combined with intermediate split thickness skin graft.
China Journal of Orthopaedics and Traumatology 2009;22(6):463-463
Adolescent
;
Adult
;
Female
;
Finger Injuries
;
surgery
;
Humans
;
Male
;
Middle Aged
;
Skin Transplantation
;
Soft Tissue Injuries
;
surgery
;
Surgical Flaps
;
Young Adult
5.Expression of Calcium/Calmodulin - Dependent Protein Kinase Ⅱ Gene in 3T3 - L1 Preadipocyte Differentiation and Regulation of Timor Necrosis Factor- ?
yong, LIU ; xi-rong, GUO ; xiao-qin, PAN ; rui, QIN ; rang-hua, CHEN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To investigate the changes of calcium /calmodulin - dependent protein kinase Ⅱ (CAMKⅡD) gene expression in 3T3 - L1 preadipocyte diffe-rentiation and TNF - ? regulation of CAMKⅡD gene expression on matured adipocytes. Methods 3T3 - L1 preadipocytes were cultured and differentiated into adipocytes. The levels of CAMKⅡD gene mRNA expression at various times were evaluated by RT-PCR. Matured adipocytes were interfered with 0.1,1.0,10.0 ?g/L TNF - ? and the levels of CAMKⅡD gene mRNA expression were evaluated. Results The levels of CAMKⅡD gene mRNA expression in 3T3 - L1 adipocytes were significantly down- regulated at first day, compared with those at zero day (P0. 05). CAMKⅡD gene mRNA expression decreased significantly at 12 hours after treatment with 10.0 ?g/L TNF-?, treatment of matured adipocytes with TNF - ? did not have any regulation role on it. Conclusions CAMKⅡD gene is involved in the differentiation of adipocytes and related to the etiology of obesity. The changes in the level of CAMKⅡD gene mRNA expression during 3T3 - L1 preadipocyte differentiation may contribute to the differentiation and adipogenesis of adipocytes. It seems that TNF - ? does not have any regulation role on the expression of CAMKⅡD genes.
6.Study on the Optimum Culture for Growth of Bacillus Subtilis B115 andthe Antibacterial Effect of B115 on Aeromonas
Zhi-Hua SHEN ; Jin-Yu SHEN ; Wen-Lin YIN ; Xiao-Yi PAN ; Ying-Lei WU ;
Microbiology 1992;0(04):-
The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.
7.Enhanced Stability of Plasmid pcDNA3.1+ within Salmonella typhimurium by Downregulation of the Ampicillin Resistance Gene Expression
Xiao-Ming ZHANG ; Xin-An JIAO ; Li-Hua TANG ; Zhi-Ming PAN ; Jin- HUANG ; Xiu-Fan LIU ;
Microbiology 1992;0(05):-
The high-copy-number plasmid pcDNA3.1+ is unstable within S almonella typhimurium. A novel plasmid pmcDNA3.1+ was constructed by removin g the promoter sequence of ampicillin resistance gene (bla gene) in plasmid pcDNA3.1+. In contrast to pcDNA3.1+, pmcDNA3.1+ was stable within Salmonel la typhimurium SL7207 in LB medium with or without ampicillin. Further experi ments showed the ?-lactamase activity of Salmonella typhimurium SL7207(pmc DNA3.1+) was apparently lowered than that of Salmonella typhimurium SL7207( pcDNA3.1+) and the high ampicillin concentration was maintained longer in LB me dium culturing Salmonella typhimurium SL7207(pmcDNA3.1+). When mice were a dministered with Salmonella typhimurium SL7207(pmcDNA3.1+) intraperitoneall y, more than 95% of Salmonella cells separated from the spleen still harbore d the plasmid pmcDNA3.1+ 7 days later; but 99% of Salmonella cells lost the plasmid pcDNA3.1+ at day 3 in mice innoculated with Salmonella typhimurium SL7207(pcDNA3.1+). By lowering the expression of bla gene, the rapid deco mposition of ampicillin in LB medium was avoided and the metabolic pressure was relieved for the host cells. This method offers a solution for the problem of t he instability of high-copy-number plasmid within Salmonella typhimurium.
8.Effect of health education on blood pressure control in hypertensive population from a community of Wuzhou city
Shaojuan QIU ; Can XIAO ; Hua LI ; Yumei HUANG ; Fengzhen HUANG ; Jianneng LUO ; Zhiping PAN
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(3):273-275
Objective:To study effect of health education on blood pressure control in patients with hypertension through community survey and perform health education.Methods:Residents from Datang street community re-ceived systematic sampling and questionnaire.A total of 490 patients with hypertension detected during census were regarded as subjects,and they were randomly divided into routine treatment group (n=240)and health education group (n=250).Health education group received health education,and effects were compared between two groups. Results:Compared with routine treatment group after intervention,there were significant rise in awareness rate of preventive and therapeutic knowledge [diagnostic value of hypertension (49.17% vs.58.40%),risk factors of hy-pertension (30.41% vs.58.00%),antihypertensive medication (58.75% vs.79.60%)]and control rate of blood pressure (23.33% vs.60.80%),and significant reduction in blood pressure [(147.68±13.97)/(89.30±12.43)mm-Hg vs.(143.57±13.53)/(85.73±11.54)mmHg]in health education group,P<0.05 or<0.01. Conclusion:Carry-ing out targeted health education possesses active significance for prevention and treatment of hypertension.
9.Leptin expression in rats model with traumatic spinal cord injury and femoral fracture
Lei WANG ; Linjuan LIU ; Li XIAO ; Xiang YAO ; Hua DING ; Zhanpeng PAN
The Journal of Practical Medicine 2015;31(22):3688-3690
Objective To observe serum and callus leptin expression in rats model with fracture and traumatic spinal cord injury (SCI). Methods 72 male SD rats were randomized equally into 4 groups: control, SCI group, fracture group, and fracture/SCI group. Rats were sacrificed at 7, 14, 21 and 28 days after fracture/SCI. Serum leptin was detected by radioimmunoassayat 1, 7, 14, 21 and 28 days, and callus formation was measured radiologically at 14, 21 and 28 days. Callus leptin was analyzed with immunohistochemistry. Results Serum leptin in the fracture group, SCI group and combined fracture/SCI group were all higher than in control group at the 1, 7, 14 and 21 day time-point (P < 0.05). Serum leptin in the combined fracture/SCI group was significantly higher than the fracture group at 7, 14 and 21 days (P < 0.05), and higher than SCI groups at 14 and 21 days after operation (P < 0.05). The percentage of leptin-positive cells in the fracture/SCI callus, and callus volume was significantly higher than the fracture-only group (P < 0.001). Conclusions Leptin expression increases in the recovery process after SCI, and the recovery of fracture becomes sooner.
10.Study on cytokines and dendritic cells involving immunologic inhibition of rat high-risk corneal transplantation induced by SEB
Xiao-Bo TAN ; Zhi-Qiang PAN ; Yu CHEN ; Ying JIE ; Wen-Hua ZHANG ; Yu-Ying WU ;
Ophthalmology in China 1993;0(03):-
objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.