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.

Objective To build the method of computerized case classification,for the purpose of perfecting the diseases typing and classification,and supporting the management based on the quality and expenses of the disease.Methods 875 cases were defined into two types.Eight parameters were selected for non-surgery cases,namely the disease diagnosis,severity at admission,and age.For surgery cases,five parameters were defined,namely the disease diagnosis,operative quantity,and severity at admission.Then Fisher function was called into play to obtain the function descriminant equation,realizing computerized classification.Results The rate of matching was 86.2％ between computerized classification and manual classification.The high accuracy of function descriminant equation proves the satisfactory outcomes of the classification.Collusion The computerized classification is satisfactory in its outcomes,and therefore it can better quality of care and cost management of diseases in clinical practice.

Adult ; Aneurysm, Dissecting ; etiology ; Coronary Aneurysm ; etiology ; Humans ; Male ; Myocardial Infarction ; etiology ; Wounds and Injuries ; complications

OBJECTIVETo investigate the effects of hydrogen sulfide (H₂S) on oxidative stress and endoplasmic reticulum stress (ERS) in a rat model of diabetic cardiomyopathy (DCM).

METHODSThirty male SD rats were randomly divided into control group, diabetes group and treatment group( n = 10). Intraperitoneal injection of streptozotocin was utilized to establish a rat model of DCM. The rats with DCM in treatment group were intraperitoneally injected with NaHS solution. After treated for 12 weeks, the hearts isolated from rats were perfused on a langendorff apparatus. The ventricular hemodynamic parameters were measured. The ultrastructures of myocardium were observed using electron microscopy. The content of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in myocardial tissue were determined by spectrophotometry. The expressions of C/EBP homologous protein( CHOP), glucose-regulated protein 78 (GRP78) and Caspase 12 at mRNA level in myocardium were detected using RT-PCR.

RESULTSCompared with control group, the cardiac function and myocardial ultrastructure were damaged obviously in diabetic rats. In myocardial tissue, the content of MDA was increased, while the activities of SOD and GSH-Px were decreased. CHOP, GRP78 and Caspase 12 mRNA expressions were increased significantly. Compared with diabetes group, cardiac function and myocardial ultrastructure damage were improved in treatment group. The content of MDA was decreased, while the activities of SOD and GSH-Px were increased significantly. The mRNA levels of CHOP, GRP78 and Caspase 12 were increased.

CONCLUSIONH2S can protect myocardium in diabetic rats, maybe it is related to reduce oxidative stress damage and inhibition of the ERS-induced apoptosis pathway.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Cardiomyopathies ; drug therapy ; Endoplasmic Reticulum Stress ; Glutathione Peroxidase ; metabolism ; Heat-Shock Proteins ; metabolism ; Hydrogen Sulfide ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Myocardium ; ultrastructure ; Oxidative Stress ; Rats ; Streptozocin ; Superoxide Dismutase ; metabolism ; Transcription Factor CHOP ; metabolism

Analyzed the prescriptions for phlegm retention syndrome that built by Ma Peizhi by the association rules and clustering algorithm, the frequency of drug usage and the relationship between drugs could be get. And from that we could conclude the experiences for phlegm retention syndrome of Ma Peizhi of menghe medical genre. The results of the analysis were that 18 core combinations were dig out, such as Citri Exocarpium Rubrum-Eriobotryae Folium-Citri Reticulatae Pericarpium. And there were 9 new prescriptions were found out such as Aurantii Fructus-Citri Exocarpium Rubium-Eriobotryae Folium-Citri Reticulatae Pericarpium. The results of the analysis were proved that Ma Peizhi of Menghe Medical Genre was good at curing phlegm retention syndrome by using the traditional Chinese medicine of mild and light, such as the medicines of mild tonification, and clearing damp and promoting diuresis.
Algorithms ; Data Mining ; Databases, Factual ; Drug Prescriptions ; standards ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Mucus ; chemistry ; Practice Patterns, Physicians' ; Respiratory Tract Diseases ; drug therapy

Objective:To investigate effects of hydrogen sulfide (H2S) on inducible nitric oxide synthase (iNOS) in kidneys of Type 1 diabetic rats.Methods:Thirty-two male SD rats were randomly divided into four groups:A normal control (NC) group,a diabetes mellitus (DM) group,a NaHS (NaHS+DM) group,and a NaHS control (NaHS) group (n=8 per group).Type 1 diabetes was induced by a single intraperitoneal injection of streptozotocin (55 mg/kg).After successful establishment of models,the rats in NaHS+DM and NaHS groups were injected with NaHS solution (56 μmol/kg) intraperitoneally.Eight weeks later,the activities of total nitric oxide synthase (T-NOS) and iNOS,as well as the level of nitric oxide (NO) were detected in serum and renal tissues,respectively,The activity of glutathione peroxidase (GSH-Px) was determined in renal tissues,The ultrastructures of renal tissues were observed by transmission electron microscope,The protein expression ofiNOS in renal tissues was detected by Western blot.Results:Compared with the NC group,there was no significant difference in the various indexes in the NaHS group (P＞0.05).However,in the DM group,the activities of T-NOS and iNOS,and the level of NO were all increased significantly in serum and renal tissues,while the activity of GSH-Px was decreased in renal tissues.Under the electronic microscope,the thickening of the glomerular capillary basement membrane,the proliferation of mesangial matrix,and the foot fusion were observed.The protein expression ofiNOS was increased obviously in renal tissues in the DM group (P＜0.01).Compared with the DM group,the activities of T-NOS and iNOS and the level of NO were all decreased in serum and renal tissues,while the activity of GSH-Px was increased in renal tissues in the NaHS+DM group (P＜0.01).The renal ultrastructural damages were ameliorated obviously.The protein expression ofiNOS was decreased significantly (P＜0.01).Conclusion:H2S exerts a protective effect on kidney injury in type 1 diabetic rats.The mechanism might be related to inhibition of iNOS activity and protein expression,in turn leading to reduction of NO content in renal tissues.

Objective To investigate the drug combination of mutual restraint which is one of the application of seven different combination in drug in traditional Chinese medicine. Methods With the methods of association rules and complex system entropy clustering, we analyzed the drug combinations of mutual restraint that were collected from Compendium of Material Medica, to obtain the frequence of the single drug and drug combination of mutual restraint and the association rules between them. Results The single drugs with higher frequency are Rhizoma Coptidis, Fructus Crotonis, Rhei Radix Et Rhizoma, Azurite, Rhizoma Acori Tatarinowii, etc. The drug combinations with higher frequency are Azurite and Artemisia capillaries, Fructus Crotonis and Rhizoma Coptidis, Fructus Crotonis and Soybean etc. The associative drug combinations which confidence is 1 include “Indigo juice → Fructus Crotonis”,“Indigo juice → Rhizoma Coptidis”, etc. Conclusion The drug combinations of mutual restraint mainly include mineral drugs, antipyretics, purgatives, and toxic drugs.

Objective To observe the relationship between serum level of H-FABP before percutaneous coronary intervention (PCI) and prognosis in patients with ST-segment elevation myocardial infarction (STEMI). Methods A total of 104 cases were recruited in this study. PCI was performed 12 h after onset. The concentration of H-FABP was detected before operation. Gensini score system was used to calculate the integral of all patients after coronary angiography. According to Gensini score, patients were divided into three groups, 34 cases in group A (8≤Gensini scores<41), 34 cases in group B (41≤Gensini scores<64) and 36 cases in group C (Gensini scores≥64). Indicators were analyzed statistically including systolic blood pressure (SBP), diastolic blood pressure (DBP), smoking history, H-FABP, left ventricular ejection fraction (LVEF), total cholesterol (TC), triglycerides (TG), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), fasting blood sugar, diabetes (DM) and hypertension. The patients were followed up for 12 months after operation. According to the occurrence of major adverse cardiovascular events (MACE), patients were divided into MACE group and non-MACE group. The related factors of two groups were statistically analyzed. The multivariate Logistic regression analysis was used to screen the risk factors of MACE for patients with STEMI after emergency PCI. Results There is a gradually increasing tendency in total cholesterol levels among three A, B and C groups (P<0.05). LVEF values were lower in B group and C group than that of A group (P<0.05). There were no significant differences in levels of SBP, DBP, H-FABP, TG, HDL-C, LDL-C, fasting plasma glucose and smoking history, DM, and hypertension between three groups (P>0.05). In 12-month follow-up, the proportion of STEM combined with DM, the time from onset to PCI, the level of TC, the level of H-FABP before operation and Gensini score were significantly higher in MACE group than those of non-MACE group (P<0.05). Logistic regression analysis showed that STEM combined with DM, higher serum level of H-FABP before operation were risk factors of MACE in 12-month after operation (P<0.05). Conclusion For patients with acute STEM combined with diabetes and elevated preoperative serum level of H-FABP, be alert to the occurrence of MACE in 12-month after PCI.

Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) coupled with a molecular network analysis strategy was used to identify the chemical constituents of the stem bark of two kinds of asparagus. The chemical constituents were identified by determining an accurate molecular weight, the fragmentation pathway, and comparison with the mass spectrometry data from the references. A molecular network was established based on the similarity of MS/MS fragmentation patterns. A total of 107 compounds were identified or tentatively deduced, which included 46 saponins, 13 flavonoids, and 48 other compounds. The chemical compounds identified in the stem bark of white and green asparagus differed greatly: the white asparagus was rich in saponins, while the green asparagus was rich in flavonoids. In conclusion, the chemical constituents of asparagus stem bark were characterized rapidly using UPLC-Q-TOF-MS/MS and molecular network analysis, with 10 compounds and 45 targets determined from the HIT 2.0 herbal ingredients' targets platform. This work will provide a theoretical basis for the resource utilization of asparagus.