BACKGROUND: Gene transfer to study the effect of a exogenous gene on cells is a commonly used technique for molecular biology. Compared with viral vector,pcDNA4.0-TGF-?1 eukaryotic expression plasmid transfection has no inherent toxicity or cytotoxicity,with high safety. OBJECTIVE: To construct and identify eukaryotic expression plasmid carrying recombined TGF-?1 by cloning transforming growth factor (TGF)-?1 gene. DESIGN,TIME AND SETTING: The in vitro cell experiment was performed at the Opening Laboratory of Key Clinical Medical Sciences,Henan Provincial High-learning School from March 2007 to February 2008. MATERIALS: Clean healthy Sprague Dawley rats aged 2 months,of both gender,were used for isolating RNA from cells. METHODS: The TGF-?1 mRNA was extracted from the rat bone marrow cells,and amplified by reverse transcription-polymerase chain reaction (RT-PCR),and cloned into the pGEM-T-vector and identified by PCR. Then TGF-?1 gene was subcloned into pcDNA4.0 plasmid vector. The inserting DNA sequences were analyzed. MAIN OUTCOME MEASURES: Expression of TGF-?1 cDNA,pGEM-T-TGF-?1 and recon pcDNA4.0-TGF-?1. RESULTS: There was an obvious electrophoresis strip of TGF-?1 cDNA by RT-PCR. The obtained pGEM-T-TGF-?1 was confirmed correct with PCR amplification and sequence identification. Recombinant eukaryotic expression plasmid pcDNA4.0-TGF-?1 was obtained and identified by digestion. The inserted sequences were conformity to the designed sequences. CONCLUSION: Rat TGF-?1 gene has been successfully cloned,and recombinant TGF-?1 eukaryotic expression vector has been constructed successfully.

OBJECTIVE: Animal experiments prove that alcohol may cause adipo-accumulation in femoral head and then induce osteonecrosis,but its mechanism of action is not known. NIH 3T3 fibroblast is a kind of pluripotent stem cell.There are not relative reports on the effect of alcohol on NIH 3T3fibroblast at present.OBJECTIVE: To observe the effect of alcohol on the differentiation of NIH 3T3 fibroblast into adipocyte and the influence of that on the expression of adipogenic transcription factor-peroxisome proliferator activated receptor gamma (PPAR-γ).DESIGN: Single sample observation.SETTING: Department of Biochemistry, Basic Medical College,Zhengzhou University and Department of Orthopaedics, First Affiliated Hospital, Zhengzhou University.MATERIALS: The experiment was completed in the Laboratory of Department of Biochemistry, Basic Medical College, Zhengzhou University from April 2002 to June 2003.NIH 3T3 fibroblasts were provided by the Laboratory of Department of Orthopaedics,University of Virginia Hospital.METHODS: The NIH 3T3 fibroblasts were divided into 6 groups randomly.The alcohols (volume fraction was 0.998) with the dosage 0(control group),0.03,0.06,0.09,0.15,0.21 mol/L were added respectively to the NIH 3T3 fibroblasts daily.The mixtures were cultured for 14 days and then stained with Sudan Ⅳ .The percentage of adipocyte was determined with image analysis software of computer,Image Pro Plus 4.1.The expression of PPAR-γ mRNA was determined with the technology of reverse transcription-polymerase chain reaction.alcohol and control groups.RESULTS: The cells were processed with the alcohols of progressive inpercentages of adipocytes in all the alcohol groups were 11.9% ,23.8%,28.2% ,31.1% ,42.6% respectively and 1.2,2.5,2.9,3.2,4.4 times that in control group(9.6%) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant (P ＞ 0.05),but the differences between the other 4 groups and control group,0.03 mol/L alcohol ues of PPAR-γ mRNA to 18S gene in the cells of all the groups were 0.218,0.411,0.486,0.473,0.453 respectively and were 1.1,2.1,2.5,2.4,2.3times that in control group (0.197) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant(P ＞ 0.05),but the differences between the other 4 groups and control group and 003 mol/L alcohol group were significant extremely(P ＜ 0.001).CONCLUSION: Alcohol can induce differentiations of a large quantity of NIH 3T3 fibroblasts into adipocytes directly, which might be one of the factors of lipotrophy in bone marrow wheri alcohol-induced osteonecrosis happens.

Objective To build unattended in-patient deep venous thrombosis (DVT) assessment system through literature review and clinical practice.Methods Making out the first items through looking up literature review and guidelines and expert consultation,the unattended in-patients were chosen in hospital to scan the items,power the score and set degrees,then different suggestions were made for different degrees.Results The in-patient deep venous thrombosis assessment system included 13 items and 4 degrees.The value was increasing with the danger of deep venous thrombosis,and seven was the point of the curve of deep venous thrombosis incidence.The higher the score was,the higher the rate of composition,and the danger will be more probable.Hence we should pay more attention on patients who had two or more items.Conclusions The study assesses the items quantitatively and make suggestions according to different evaluation levels.The system help nurses to prevent deep venous thrombosis in early stage and make reference for clinical practice.

Objective To find a method to inherit the experiences of famous senior TCM physicians,through verification and inheritance research of Professor Tian Cong-huo's clinical experience on degenerative gonarthrosis treatment.Methods By means of database of famous senior TCM physicians and various data explore methods,we performed multi-group controlled verification on Professor Tian Cong-huo's acupuncture prescription on degenerative gonarthrosis,in order to investigate the experience thoroughly.Results The three groups treated by Professor Tian Cong-huo's acupuncture prescription got similar therapeutic effects,and patients' symptoms have been relieved remarkably.Compared with the baseline,the scores of the knee joint ache in the group of Professor Tian,Successor,Senior and Control were decreased 1.20 (t=9.00,P＜0.05),1.13 (t=15.76,P＜0.01),1.1 (t=15.99,P＜0.05) and 1 (t=10.42,P＜0.05),respectively.Significant improvements of pain relieves had been achieved in all groups.No significant difference was observed among the previous three groups(F=2.64,P＞0.05),which had obvious improvement effect in comparing with the control group(t=3.81,P＜0.01).Conclusion demonstrated that the group of Professor Tian,the successor,and the senior each did better in knee pain-relief than the control group.Conclusion Confirmed by our research,this acupuncture prescription proves to be repeatable,safe,reliable and stable.Perfoming inheritance and verification research of famous and senior TCM physicians could help juniors learn and investigate their predecessors academic thoughts and connotation,proceed to the next step,inherit the precious experience through verification and combination of theory and practice.

Objective To study the effect of consciousness restoring needling method combined with Jingjin needling method for shoulder-joint semiluxation patients after stroke.Methods The 108 patients were divided randomly into the treated group(n=58) and the control group(n=50).The treated group was treated with consciousness restoring and Jingjin Needling while the control group treated with consciousness restoring only.The therapeutic effect,shoulder joint movement,and severity of nervous impairment of both groups were observed.Results There were significant differences between the two groups in total effective rate and improvement of joint movement and nervous function(P

Objective To explore the relationship among the severity of coronary disease, c-reaction protein (CRP) and mild renal dysfunction. Methods 64 patients underwent coronary angiography were divided into two groups according to their serum creatinine (cr) levels: normal renal function(cr

Objective To evaluate the effect of fast track surgery nursing on percutaneous lumbar diskectomy. Methods A total of 126 patients were randomly divided into the control group and the experimental group,the control group received traditional nursing plan, the experimental group was given fast track surgery nursing plan. The nursing effect was observed. Results Postoperative anal exhaust time, feeding time, walking time, length of hospital stay ofthe experimental group were (1.61±0.71) days, (2.01±0.71) days, (4.70±2.12) days, (12.72±2.07) days, whichwere significantly shorter than those of the control group, which were (3.37±0.93) days, (2.27±0.63) days, (8.40±3.21) days, (15.81±2.14) days, the difference between two groups were statistically significant (t=4.012-6.068, P<0.05). The incidence of complications was 11.1%(7/63) in the experimental group, which was lowercompared with the control group, 23.8% (15/63), the difference between two groups was statistically significant(χ2=6.639, P < 0.05). Curative effect of the experimental group was 90.4% (57/63), which was significantly higher than that of the control group, 85.7% (54/63), the difference between two groups was not statistically significant (χ2=1.460,P>0.05). The Rdand Morris Questionnaire score of the experimental group was 21.67±3.48, which was higher than that of the control group(14.43 ± 2.01), the difference between two groups was statistically significant(χ2=6.077, P <0.05). Conclusion Fast track surgery nursing can promote the patient's postoperative rehabilitation ,reduce the postoperative complications, shorten the length of hospital stay and improve the quality of life.

Objective To investigate the influence of fast track surgery (FTS) on insulin resistance indexes and inflammatory reaction in colorectal cancer patients who received laparoscopic surgery.Methods A total of 62 patients were randomly divided into control group (n=31) or FTS group (n=31).The postoperative first flatus and defecation time, postoperative hospital stay, hospitalization expenditure, and postoperative complications were recorded.Insulin resistance indexes and inflammatory reaction were measured before operation (T0) as well as on the 1st, 3rd, and 7th postoperative days (T1, T3, and T7), including fasting blood-glucose (FBG), fasting insulin (FINS), serum level of interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor (TNF-α).The insulin resistance index (HOMA-IR) was calculated at the same time points.Results The postoperative time of flatus and defecation in the FTS group was significantly shorter than that in the control group (t=3.088, P=0.003;t=4.227, P=0.000), the postoperative hospital stay and total hospitalization expenditure were significantly lower in FTS group (t=3.937, P=0.000;t=3.478, P=0.003).No significant difference was found in the complication rate between the two groups (χ2=0.267,P=0.605).The inflammatory indexes and insulin resistance indexes in the two groups at T1 and T3 time points were higher than those at T0 time points (P<0.05).Compared to the FTS group at T1 and T3 time points, the levels of IL-6, CRP, TNF-α, FINS, and HOMA-IR in the control group increased more obviously (P<0.05), but the elevation blood FBG was not obvious (T1: t=0.870, P=0.388;T3: t=0.870, P=0.388).There were no significant differences in IL-6 and TNF-α in the two groups between the T7 and T0 time points (P>0.05), but the levels of CRP, FBG, FINS, and HOMA-IR at T7 time points were significantly higher than those at the T0 time points in the two groups (P<0.05).The levels of IL-6, TNF-α, FBG, FINS, and HOMA-IR at T7 had no significant differences between the two groups (P>0.05), but the level of CRP in the control group was significantly higher than that in the FTS group (t=-4.527, P=0.000).Conclusion FTS may promote the early recovery of gastrointestinal functions, reduce stress reactions and postoperative insulin levels, thus being conducive to fast rehabilitation in colorectal cancer patients with laparoscopic surgery.

Objective To explore the effect of progesterone on the rate of neuronal apoptosis and nitric oxide(NO) level in the cortex and hippocampus tissue of newborn rats with hypoxic-ischemic encephalopathy(HIE).Methods Thirty 7-day-old neonatal rats were randomly divided into 3 groups:sham-operated group,hypoxic-ischemic(HI) group and pretreatment group.Rats in HI group and pretreatment group were subjected to left common carotid artery ligation,then were exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for up to 2.5 h to establish HIE model.Progesterone was injected intraperitoneally into rats in the pretreatment group respectively for 30 minutes before hypoxia,and solution was injected into the sham-operated group and HI group.All rats were killed at 24 h after operation.The neuron apoptosis was identified and analyzed by flow cytometry.Nitrate/nitrite was assayed to represent nitric oxide content of brain tissues.Results The ratio of neuronal apoptosis and NO contents in cortex and hippocampus tissue in HI group [(10.09?0.36)%,(12.32?0.28)%,(51.36?9.71) ?mol/L,(52.34?4.26) ?mol/L] were significantly higher than those in sham-operated group [(2.49?0.23)%,(2.58?0.26)%,(18.16?6.24) ?mol/L,(19.28?3.58) ?mol/L)](P_a

Inheirted metabolic disodrers(IMD)involves in multiple substance dysbolism,which usually results in irreversible neurological lesions because of various categories and complicated clinical manifestations.In resent years,IMD became one of the hot spots in medical domain around the world,original diagnostic technique and management progressed unceasingly.This paper provides an overview of the traditio-nal detection and treatment about IMD,and reviews the new techniques such as gene analysis,gene chip,organ transplantation and enzyme replacement therapy at the same time.