BACKGROUND: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is not applicable for clinical fast detection or the detection analysis of multiple genes on a large sample basis for candidate genes of mental diseases, while polymerase chain reaction-sequence specific primer (PCR-SSP) method can make up for these deficiencies.OBJECTIVE: To establish a PCR-SSP method with multiple gene polymorphism, and popularize its application in the molecular study of mental diseases.DESIGN: Observational trial.SETTING: Department of Molecular Biology and Department of Pathophysiology at Xinxiang Medical College.MATERIALS: The experiment was carried out in the Department of Molecular Biology at Xinxiang Medical College from December 2005 to October 2006. A total of 200 blood samples from patients with mental disease were provided by the Second Affiliated Hospital of Xinxiang Medical College, 0.5 mL each sample. All the patients were informed and agreed with the experiment.METHODS: PCR-SSP was used to analyze the polymorphism of the following genes: variation of tumor necrosis factor (TNF)-α-308A/G and -238G/A, variation of interleukin (IL)-6-174G/C, variation of the 188th base pair (C/T) of CYP2D6 *10B exon, variation of the 472nd base pair (G/A) of COMT gene, and variation of the 941st base pair (G/T) of the CDNA sequence of the 8th exon of MAOA gene.MAIN OUTCOME MEASURES: Gene polymorphism of blood samples analyzed with PCR-SSP.RESULTS: The detected genotypes for TNF-α-308NG included G/G and A/G; the detected genotypes for TNF-α-238G/A included G/A, G/G and A/A; the detected genotypes for IL-6-174G/C were all G/G homozygotes; the detected genotypes for the 188th base pair (C/T) of CYP2D6*10B exon included C/C, C/T and T/T; the detected genotypes for the 472nd base pair (G/A) of COMT included A/G, A/A and G/G; the detected genotypes for the 941st base pair (G/T) of CDNA sequence of the 8th exon of MAOA included T/G, T/T and G/G.CONCLUSION: PCR-SSP is suitable for the polymorphism analysis of the mononucleotide site variation of multiple genes on a large sample basis, which is fast and accurate with low cost and is worthy to be popularized in the molecular study on mental diseases.

Objective To establish an approach of polymerase chain reaction-sequence special primers(PCR-SSP) to perform polymorphism analysis for multiple genes simultaneously,in order to be used in clinical analysis.Methods The optimized PCR-SSP approach was used to analyze the polymorphism of the following genes:mutation of-308A/G and-238G/A in tumor necrosis factor-?((TNF-?)),-174G/C in interleukin-6(IL-6).Results Polymorphism analysis of multiple genes and many clinical samples could be simultaneously performed with one PCR program,showing clear genotype,quick and accurate genotype.Conclusion The optimized(PCR-SSP) is suitable for polymorphism analysis of the large-sample polygenic mononcleeotide point mutation,and it can be widely applied in clinical test for low cost,quickness and accuracy.

OBJECTIVETo evaluate the curative effect of Qingkailing Injection (QKL) for treatment of children's respiratory syncytial virus pneumonia of phlegm-heat obstructing Fei syndrome pattern (SVP-PH) depending upon main symptom assessment.

METHODSA Chinese-Western medicine comparative trial was conducted on 206 children with SVP-PH in two groups treated with Ribavirin injection plus compound guaiacol potassium sul-fonale oral solution (as control group) and QKL injection plus Ertong Qingfei oral liquid (as treated group) respectively, for 10 days. The curative effectiveness on four main symptoms (fever, cough, sputum and short breath) were evaluated at different time-points.

RESULTSThe effectiveness in the treated group at various time-points was superior to that in the control group, showing the earlier initiating time (on the 4th day) and the preponderances on cough and sputum ran all through the whole course.

CONCLUSIONChinese medicine shows a multi-target effect in treating children's SVP-HP.

Child ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Guaiacol ; analogs & derivatives ; therapeutic use ; Humans ; Infusions, Intravenous ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Pneumonia, Viral ; drug therapy ; Respiratory Syncytial Virus Infections ; drug therapy ; Ribavirin ; therapeutic use

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the presence of abnormal metabolism in the thalamus and hypothalamus in patients with first-episode depression.

METHODSThirty drug-naive patients with first-episode depression and 30 age-matched controls were scanned with proton magnetic resonance spectroscopy ((1)H-MRS) for Naa, Cho, Cr and mI.

RESULTSCompared with the control group, the patients showed significantly reduced mI and mI/Cr of the hypothalamus, reduced mI/Cr of the left thalamus, and lowered Cho, ml, and ml/Cr of the right thalamus (P<0.05).

CONCLUSIONPatients with first-episode depression may have myo-inositol and phosphoric acid metabolism disorder in the thalamus and hypothalamus with malfunction of cellular osmotic pressure adjustment mechanism. Abnormal mI/Cr in the thalamus and hypothalamus may represent an important biochemical change in advanced patients with depression.

Adolescent ; Adult ; Case-Control Studies ; Choline ; metabolism ; Creatine ; metabolism ; Depression ; diagnosis ; Female ; Humans ; Hypothalamus ; metabolism ; Inositol ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Male ; Middle Aged ; Protons ; Thalamus ; metabolism ; Young Adult

Objective To investigate the relationship of the levels of peroxisome proliferator- activated receptor-?(PPAR-?)mRNA,MMP-9 with the severity of coronary artery lesions in patients with coronary heart disease(CHD).Methods One hundred and fifty three patients with CHD who underwent coronary angiography were admitted.The expression of PPAR-?mRNA in lymphocytes of peripheral blood was detected by using RT-PCR,the level of MMP-9 enzyme was measured by enzyme-linked immunosorent assay,and the severity of coronary artery lesions were analyzed. Results As compared with control(0.624-0.13),PPAR-?mRNA expression was significantly lower in CHD patients(0.4+0.12).Negative correlation was found between PPAR-?mRNA and the classification(r=-0.56,P＜0.01)of coronary artery lesions,so was the number of coronary artery lesions(r=-0.42,P＜0.01).MMP-9 level was significantly higher in CHD patients(1.27?0.16)?g/L than that in controls(1.21?0.05)?g/L.Positive correlation was found between MMP-9 level and the classification(r=0.36,P＜0.01)of coronary artery lesions,so was the number of coronary artery lesions(r=0.30,P＜0.01).Negative correlation was also found between PPAR-?mRNA expression and MMP-9 level.Conclusions PPAR-?is a negative regulator of coronary artery lesions and PPAR-?inhibits the activation of MMP-9.It may be a valuable method for protecting patients from the incident of coronary artery disease to activate the expression of PPAR-?and decrease the level of MMP-9.

OBJECTIVEAngiotensin II is an important pro inflammation factor in the cardiovascular system. This experiment is aimed to study the effects of angiotensin II on inducible nitric oxide synthase expression in human umbilical endothelial cells.

METHODSHuman umbilical endothelial cells were cultured in vitro and treated with angiotensin II alone or in combination with AT1, AT2 and NF-kappaB inhibitors respectively. The inducible nitric oxide synthase expressions at protein and mRNA levels were measured with Western blot and reverse transcription-polymerase chain reaction (RT-PCR), and the activity of NF-kappaB was analyzed with EMSA.

RESULTSAngiotensin II up-regulated inducible nitric oxide synthase expressions at the protein and mRNA levels at 5 h (P < 0.05), the activity of NF-kappaB was enhanced at 2 h (P < 0.05). These effects could be blocked by AT1 and NF-kappaB inhibitors but not by AT2 inhibitor.

CONCLUSIONAngiotensin II can upregulate the expression of inducible nitric oxide synthase through NF-kappaB pathway in human umbilical endothelial cells. AT1, other than AT2, play a key role in this process.

Angiotensin II ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Angiotensin II Type 2 Receptor Blockers ; Cell Line ; Endothelial Cells ; chemistry ; drug effects ; Heart Failure ; metabolism ; Humans ; NF-kappa B ; metabolism ; Nitric Oxide Synthase Type II ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Umbilical Veins ; cytology ; Up-Regulation

OBJECTIVETo objectively evaluate the clinical effect of traditional Chinese medicine in treating children's respiratory syncytial viral pneumonia (RSVP) of phlegm-heat blocking Fei syndrome (PHBFS).

METHODSA single-blinded multi-center, blocked, randomized and parallel-controlled method was adopted. The clinical study was carried out on 206 children with RSVP-PHBFS who were assigned to two groups, 108 in the test group treated through intravenous dripping of Qingkailing Injection () in combination of oral intake of Er'tong Qingfei Oral Liquid () and 98 in the control group with intravenous dripping of ribavirin injection in combination with oral intake of potassium guaiacol sulfonate oral liquid, all for 10 days. The clinical efficacy was evaluated and compared at the end of the trial from various aspects by three methods including comprehensive efficacy, post-treatment main symptoms score difference and survival analysis of the main symptoms.

RESULTSAfter treatment, in the test group, 60 patients were cured, 36 markedly alleviated, and 12 improved. In the control group, 41 were cured, 38 markedly alleviated, 18 improved and 1 unchanged. Comparison on the comprehensive efficacy between the two groups shows a better efficacy in the test group (chi(2)=4.4527, P=0.0348). Scores of the main symptoms were lowered after treatment in both groups, the difference was 22.41+/-4.99 scores in the test group and 17.61+/-6.34 scores in the control group, being more significant in the former (t=-5.99, P<0.01). Survival analysis shows that there was significant difference between the two groups in the effect initiating time on such symptoms as fever, cough, copious sputum, shortness of breath, and rales, which was earlier in the test group (P<0.01 or P<0.05).

CONCLUSIONEvaluation of the efficacy of traditional Chinese medicine in treating children with RSVP-PHBFS by using the three methods jointly could better show the objectivity of the evaluation.

Airway Obstruction ; complications ; mortality ; therapy ; Child, Preschool ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fever ; complications ; mortality ; therapy ; Hot Temperature ; Humans ; Infant ; Male ; Medicine, Chinese Traditional ; methods ; Pneumonia, Viral ; drug therapy ; mortality ; pathology ; Respiratory Syncytial Virus Infections ; drug therapy ; mortality ; pathology ; Respiratory System ; pathology ; Ribavirin ; administration & dosage ; Single-Blind Method ; Survival Analysis ; Syndrome ; Treatment Outcome

OBJECTIVETo study the effects of insulin on the proteolysis of cultured rabbit skeletal muscular myotubes in vitro, and their possible mechanisms.

METHODSMuscles of lower limbs of juvenile rabbits were isolated for tissue-block culture. After passage, myoblasts were formed and fused into myotubes. Then the protein in myotubes was radiolabelled with L-[ 3,5-3H] tyrosine. The myotubes were cultured in DMEM medium containing 100 nmol/L insulin (n = 24, group B) , 100 nmol/L dexamethasone (n = 24, group C) , 100 nmol/L insulin and 100 nmol/L dexamethasone (n = 24, group D) , no insulin or dexamethasone (n =24, group A), respectively. Twenty-four hours after culture, the L-[3,5-3H] tyrosine content in culture medium and cells were determined, and the degradation rates of protein were calculated. The mRNA expressions of ubiquitin and protease C2 subunit were determined by Northern blot.

RESULTSThe degradation rates of myotube protein in group A(0. 38+/-0.04) was obviously lower than that in group C (0.50+/-0.03, P <0.01), but it was obviously higher than that in group B(0. 35+/-0.03, P <0.05). Though the degradation rates of myotube protein in group D (0.41+/-0. 03) was evidently lower than that in group C ( P < 0.01) , it was still higher than that in group A( P < 0.05 ). The mRNA expressions of ubiquitin and protease C2 subunit in group A ( the scale: 2. 4 kb ubiquitin was 0. 82+/-0. 15, 1. 2 kb ubiquitin was 0. 60+/-0. 10, C2 subunit was 0. 75+/-0. 16) was obviously lower than that in group C ( the scale: 2.4 kb ubiquitin was 2. 15+/-0. 23, 1.2 kb ubiquitin was 1.50+/-0. 14,C2 subunit was 1.50+/-0. 13 , P <0. 01) , but it in group D was lower than that in group C (the scale: 2. 4 kb ubiquitin was 1. 25+/-0. 17, 1. 2 kb ubiquitin was 0. 85+/-0. 09, C2 subunit was 0. 90+/-0. 15, P <0. 01) , and it was similar to that in group B (the scale: 2.4 kb ubiquitin was 0. 85+/-0.07, 1.2 kb ubiquitin was 0. 65+/- 0. 12, C2 subunit was 0. 76 +/-0. 09, P > 0. 05).

CONCLUSIONThe effects of insulin on the activity of ubiquitin-proteasome pathway and the proteolytic rate in normal myotubes were relatively weak. However, insulin can significantly inhibit the effects of dexamethasone on the gene expressions of ubiquitin system and the proteolytic rate in myotubes, but the mechanism needs further research.

Animals ; Cells, Cultured ; In Vitro Techniques ; Insulin ; pharmacology ; Male ; Muscle Fibers, Skeletal ; drug effects ; metabolism ; Muscle Proteins ; metabolism ; Rabbits ; Ubiquitin ; metabolism