【Objective】To explore the optimal conditions in fingerprinting (APHDF).【Methods】The human DNA fingerprints were detected by APHDF.A pair of short primers was used for amplification.The experimental conditions including template,Mg2+,deoxyribonucleotides,and parameters of cycle,were optimized.【Results】The template DNA should be abstracted freshly and the concentration should be ranged from 50～550 mg/L.The best concentration of Mg2+was 5.0 mmol/L.The deoxyribonucleotides concentration was optimal at 0.2 mmol/L.The PCR cycling parameters were as follows :The denaturing temperatures,annealing temperatures and extension temperatures were 94 ℃ and 90 ℃ for 30 s,43 ℃ and 48 ℃ for 40 s or 50 s,and 72 ℃ for 1 min or 80 s,respectively.【Conclusion】The optimal conditions of the experiment are obtained,with good reproducibility and high specificity.Therefore,this method can be widely applied in practice.

This study was aimed to diffuse the lung and promote urination for the observation of cardiopulmonary re-lated index change of cor pulmonale rats to further explore the treatment effect on the pulmonary function, pulmonary arterial hypertension (PAH) and the influence of right heart hypertrophy of rats, in order to further illustrate the ef-fect of diffusing the lung and promoting urination for cor pulmonale. Sixty Wistar rats were randomly divided into the control group, model group and Xiao-Qing-Long decoction (XQLD) group with 20 rats in each group. The AniRes2003 animal lung function analysis system was applied to measure the pulmonary function of rats. And the multi-guide physiological recorder was used in the recording of the pulmonary artery pressure of rats. The conven-tional weighing method was applied to calculate and obtain the change of right heart hypertrophy. The results showed that compared to the control group, symptoms in the model group became severe obviously, which include reduced activity, slow movement and occasional airway sputum sound, and the right heart hypertrophy index of the model group increased obviously (P < 0.01). Compared to the model group, the pulmonary function and pulmonary artery pressure of the XQLD group have obvious difference (P< 0.05). It was concluded that to diffuse the lung and pro-mote urination can effectively improve the pulmonary function, PAH and the right heart hypertrophy index of rats with cor pulmonale (fluid retention). The effect of this method is definite in the treatment of cor pulmonale.

OBJECTIVETo study the proportion and mechanism of relieving asthma of drug partnership comprising herbal Ephedrae & Pheretima.

METHODTo study relaxant effect on 10 micromol x L(-1) carbachol (CCh) and 10 micromol x L(-1) histamine (His) precontracted isolated tracheal rings and lowering effect on short-circuit current (Isc) increase induced by 10 micromol x L(-1) CCh with 3 proportions of 1:1, 1:3, 1:9 extract.

RESULT1:3 proportions dose-dependently relaxed CCh-precontracted isolated tracheal rings, IC50 of 1:1, 1:3 is 7.5, 15 mg x mL(-1) respectively, 1:9 could not produce 50% inhibition effect on CCh-evoked contraction; 3 proportions also dose-dependently relaxed His-precontracted isolated tracheal rings, IC50 of 1:9, 1:3 and 1:1 is 0.19, 0.61, 1.8 mg x mL(-1) respectively. On the other hand,the orders potency of the decrease effect on CCh-evoked short circuit current increase is 1:3 > 1:1 > 1:9. The difference is not significant (P < 0.05).

CONCLUSIONHerbal Ephedrae & Pheretima had tracheal muscle relaxant and epithelium ion secretion inhibition effect, its mechanism of relieving asthma involved anti-CCh and anti-His effect 1:3 was the most appropriate dosage ratio in the anti-asthmatic drug partnership.

Animals ; Anti-Asthmatic Agents ; administration & dosage ; pharmacology ; Asthma ; physiopathology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ephedra sinica ; chemistry ; Guinea Pigs ; Histamine Antagonists ; pharmacology ; In Vitro Techniques ; Male ; Materia Medica ; administration & dosage ; isolation & purification ; pharmacology ; Muscle Relaxation ; drug effects ; Muscle, Smooth ; drug effects ; Oligochaeta ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo look for the evidences of motilin receptor expression on interstitial cells of Cajal (ICC) of the rabbit.

METHODSmooth muscle segments with ICC were isolated from the small intestine of 10-day old rabbits. The tissue segments equilibrated in Ca(2+)-free Hanks' solution were dispersed with an enzyme solution containing collagenase type II and then Ficoll density centrifugation was used to dissociate ICC. The cells were suspended and cultured in the M199 medium. The c-kit antibody was applied to distinguish the cultured ICC. The motilin receptor was identified by immunocytochemical assay with GPR38 antibody, c-kit antibody and hoechst 33342 combined to label ICC. Cells cultured for a few days were sorted for ICC with c-kit stained green fluorescent through flow cytometry. The total RNA and proteins extracted from the sorted ICC were respectively used to verify motilin receptor on the ICC by reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting.

RESULTWe had successfully dissociated and cultured ICC of rabbit small intestine in vitro. Fluorescent staining with c-kit antibody confirmed that the culture ICC was successful. Triple-labeled immunofluorescent staining had detected the motilin receptor on membrane of ICC. Flow cytometry analysis showed that the ratio of c-kit positive cell in the cultured cells was 64.3%. The number of sorted ICC was 6.7 × 10(5) and 5.6 × 10(6). The results of RT-PCR and Western blot confirmed that the ICC had motilin receptor expression.

CONCLUSIONOur study demonstrated presence of motilin receptor on ICC of the rabbit. The present results may suggest that ICC play an important role in gastrointestinal movement induced by motilin.

Animals ; Cells, Cultured ; Interstitial Cells of Cajal ; metabolism ; Intestine, Small ; cytology ; Rabbits ; Receptors, Gastrointestinal Hormone ; metabolism ; Receptors, Neuropeptide ; metabolism

OBJECTIVETo detect serum HER-2 oncoprotein levels in patients with operable and metastatic breast cancers, and to study the correlations between serum HER-2 level and lymph node status as well as other clinical parameters.

METHODSA total of 120 women were studied consisting of 10 healthy volunteers, 31 benign breast disease, 53 operable breast cancer, and 26 metastatic breast cancer patients. The levels of serum HER-2 were measured using an enzyme-liked immunosorbent assay (ELISA).

RESULTSThe mean serum HER-2 levels were 9.6 +/- 1.5 ng/mL in healthy volunteers, 11.9 +/- 1.6 ng/mL in benign breast disease, 13.2 +/- 4.2 ng/mL in operable breast cancer, and 30.5 +/- 30.8 ng/mL in metastatic breast cancer patients. The former is much lower than the latter three (P = 0.02, 0.001, 0.03, respectively). If using 15 ng/mL as a normal baseline, elevated serum HER-2 levels were observed in none of the healthy volunteers as well as patients with benign disease, but in 18.9% (10/53) operable breast cancer patients and 61.5% (16/26) metastatic patients. In patients with operable breast cancer, there was a positive correlation between serum concentrations of HER-2 and the size of primary tumor (P < 0.05), whereas there was no correlation between serum concentration and axillary lymph node or estrogen receptor status. In patients with metastatic disease, there was no correlation with site of metastases (P > 0.05).

CONCLUSIONSerum HER-2 level was strongly correlated with tumor loads and clinical stages, thus acting as a promising predictor of cancer recurrence in breast cancer patients.

Biomarkers, Tumor ; blood ; Breast Neoplasms ; blood ; pathology ; Female ; Humans ; Liver Neoplasms ; chemistry ; secondary ; Lung Neoplasms ; chemistry ; secondary ; Lymph Nodes ; pathology ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Receptor, ErbB-2 ; blood ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

OBJECTIVETo establish a RP-HPLC method for the determination of swertiamarin, sweroside, gentiopicrin and oleanolic acid in different parts of Swertia pseudochinesis.

METHODA Zorbax SB-C18 (4.6 mm x 250 mm, 5 microm) column was used with acetonitrile-water (10:90) and methnol-water(86:14) at detection wavelengths of 238 nm, 246 nm, 274 nm and 207 nm for swertiamarin, sweroside, gentiopicrin and oleanolic acid respectively. The flow rate was 1.0 mL x min(-1) and the column temperature was 25 degrees C.

RESULTAll of the compounds were based--isolated. The linear ranges of swertiamarin, sweroside, gentiopicrin and oleanolic acid were 0.068 9-0.344 4(r = 0.999 2) , 0.001 1-0.014 0 (r2 = 0. 999 8), 0.001 1-0.013 4 (r2 = 0.999 9) and 0.001 1-0.008 8 mg x mL(-1) (r2 = 0. 999 6), respectively.

CONCLUSIONThe method is simple and accurate, which can be used for quality control of S. pseudochinesis.

Chromatography, High Pressure Liquid ; methods ; Flowers ; chemistry ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Oleanolic Acid ; analysis ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Pyrones ; analysis ; Reproducibility of Results ; Swertia ; chemistry ; Triterpenes ; analysis

OBJECTIVESThe investigation of the testicular volume, the penis length and the T, FSH, LH, PRL levels in serum were taken in 289 adolescent males to provide the valuable data for andrology.

METHODSThe adolescent males were grouped according to their age. The testicular volume was measured with testicular model and the T, FSH, LH, PRL levels in serum were determined by immunoenzymetric assay.

RESULTSThe male sexual development was rapid from age 11 to 16 and close to that of adult at age 18. Serum PRL of adolescent males was higher than that of adult males.

CONCLUSIONSThe age 11 to 16 is a period of rapid growth in sexual maturation. PRL may play an important role in sexual maturation.

Adolescent ; Body Height ; Body Weight ; Gonadal Steroid Hormones ; blood ; Humans ; Male ; Penis ; physiology ; Testis ; physiology

Objective To explore the risk factors for lower extremity amputation in patients with diabetic foot.Methods The clinical data of 1 771 patients with diabetic foot at the Air Force General Hospital of PLA from November 2001 to April 2015 were retrospectively analyzed.The patients were divided into the non-amputation and amputation groups.Within the amputation group , subjects were further divided into the minor and major amputation subgroups.Binary logistic regression analyses were used to assess the association between risk factors and lower extremity amputation.Results Among 1 771 patients with diabetic foot , 323 of them ( 18.24%) were in the amputation group ( major amputation: 41; minor amputation:282 ) and 1 448 ( 81.76%) in the non-amputation group.Compared with non-amputation patients, those in the amputation group had a longer hospital stay and higher estimated glomerular filtration rate(eGFR)levels.Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), C-reaction protein (CRP), ESR, ferritin, fibrinogen and WBC levels of the amputation group were higher , while hemoglobin albumin, transferrin, TC, TG, HDL-C and LDL-C were lower than those of the non-amputation group (all P<0.05 ).The proportion of hypertension ( 52.48% vs 59.98%) , peripheral vascular disease ( PAD ) (68.11% vs 25.04%), and coronary heart disease (21.33% vs 28.71%) were different between the amputation and non-amputation groups (all P<0.05).Multivariable logistic regression analyses showed that Wagner′s grade , PAD and CRP were the independent risk factors associated with lower extremity amputation in hospitalized patients with diabetic foot.Conclusion Wagner′s grade, ischemia of lower limbs and infection are closely associated with amputation of diabetic foot patients.

OBJECTIVETo investigate radiation protection and possible mechanisms of low intensity microwave on gamma-ray exposed mice.

METHODS96 healthy Kunming mice were randomly divided into the following four groups: normal control, microwave (120 microW/cm(2), 900 MHz), gamma-ray irradiation (5 Gy), combined exposure of microwave and gamma-ray (120 microW/cm(2) + 5 Gy). The microwave group and combined group were exposed to 120 microW/cm(2) microwave firstly, 1 h/d, for 14 days. Then the ionization and combined group were exposed to 5 Gy (60)Co gamma-ray irradiation on the 15th day. Animals were sacrificed on the third, 6th, 9th and 12th day after irradiation. The sternum and spleen paraffin section were produced, and the histological changes were observed. Apoptosis rate of mice splenic cells in each group was examined by flow cytometry, and serum concentration of antioxidant and lipid peroxide was detected at the same time.

RESULTSBone marrow was obviously injured either by radiation or microwave exposure, characterized by undergoing four-phase lesions, namely apoptosis-necrosis, void, regeneration and recovery phase. Compared with the gamma-ray group, the pathological changes in combined group were slighter and the recovery was quicker. The pathological injuries of spleen were similar to that of bone marrow. Injuries in the combined group were slighter than gamma-ray group. It showed that apoptosis rate of splenic cells in combined group was significantly lower on the 6th and 9th day after gamma-ray radiation (23.02% +/- 15.18%, 25.37% +/- 11.62% respectively) from FCM results. Assays of oxidative damages suggested that serum superoxide dismutase (SOD) level in combined group increased while lipid peroxide level decreased significantly (P < 0.05).

CONCLUSIONLow intensity microwave may exert protection effects on injuries induced by ionizing radiation. The underlying mechanisms might be related with suppression on the hematopoietic cells apoptosis induced by gamma-ray radiation, inhibition of oxidative damages, and thus enhanced reconstruction of the hematopoietic system.

Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; Gamma Rays ; adverse effects ; Male ; Mice ; Microwaves ; Radiation Protection

Animals ; Breath Tests ; methods ; Carbon Isotopes ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; etiology ; physiopathology ; Liver Function Tests ; methods ; Male ; Phenylalanine ; Rats ; Rats, Sprague-Dawley