Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.

Objective To observe the effects of astragaloside Ⅳ on electrocardiogram (ECG) and action potential of ventricular myocytes in guinea pigs. Methods ECG was recorded in vivo and ex vivo by using conventional ECG recording method from anesthetic guinea pigs and Langendoff perfusion model of hearts. Action potentials were recorded from isolated papillary muscles of right ventricles of guinea pigs by using microelectrode techniques. Results RR interval was prolonged by Astragaloside Ⅳ in a dose-dependent manner both in vivo and ex vivo. Astragaloside Ⅳ shortened action potential duration (APD), while had no effects on resting potential, action potential amplitude and maximal rate of depolarization. Conclusion Astragaloside Ⅳ exerts a negative chronotropic effect on heart and shortens APD of cardiac myocytes, which may be involved with calcium channels.

Objective To estimate curative effect of reconstruction of rabbit knee joint cartilage defect with the homogeneitic tissue engineered cartilages.Methods The chondrocytes were isolated and collected from articular cartilages of eight New Zealand white rabbits.The tissue engineered cartilages after culturing chondrocytes and atelocollogen for two days.Cartilage defects were created in both keen joint of twenty-six rab- bits.Complexes of chodrocytes and atelocollagen was grafted into the defect of left knee joint at once as experi- mental group,and no implantation were served as control.General and histological examination were respec- tively performed in both group at four weeks and eight weeks after surgery.Results After implantation,the defects were filled with cartilaginous tissue in experiment group,while there were only tissue in control group. Histologically,defective areas were filled with chondrocytes in experiment group,but only fibroblast in control group.Conclusion The implantation of the tissue engineered cartilages contenting with chondrocytes and atelocollogen can effectively improve reconstruction of rabbit knee joint.

OBJECTIVE: In order to explore the relationship between the expression change of cytokines and the wound age during the healing process of rats skin wound. METHODS: A assay of ELISA were performed on intravital skin wounds(after incision 0.5-168 h) to detect the dynamics expression level of IL-2, TNF-alpha. RESULTS: The level of IL-2 and TNF-alpha increased at 0.5 h after wounding, then got to a peak at 3 h and 1 h after injure respectively. Rebound of TNF and IL-2 levels were shown at 48 h after wounding, and both cytokine levels were inclined to elevating between 72 h and 168 h after wounding. CONCLUSION The cytokine level changes suggest they have a time-related expression during wounds healing process.
Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Interleukin-2/metabolism* ; Male ; Rats ; Rats, Wistar ; Skin/metabolism* ; Time Factors ; Tumor Necrosis Factor-alpha/metabolism* ; Wound Healing/physiology*

Objective To screen the optimized Buyang Huanwu Decoction (BYHWD);To verify it. Methods H2O2 was used to induce PC12 cell oxidative stress models. MTT method was used to determine the prevention effects of BYHWD at different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL) on in vitro oxidative stress cell models to define the optimized concentration. Orthogonal design was used to divide BYHWD single medicine into decomposed BYHWD groups, control group (only with DMEM), normal group (without H2O2 and medicine processing), and model group, to investigate the protective effects on PC12 cells. Optimized BYHWD was screened to decide the compatibility ratio of each medicine. MTT was used to detect the cell survival rate in each group. Middle cerebral artery occlusion was used to replicate MACO rat models. SD rats were randomly divided into sham-operation group, model group, BYHWD group and optimized BYHWD high-, medium-and low-dose groups. Each medication group was given relevant medicine for gavage. The screened results were verified. Results Compared with other decomposed BYHWD groups, the protective effects of the compatibility of Astragali Radix+Chuanxiong Rhizoma+Pheretima on PC12 cells was the best (P<0.05), which was nearly equaled to BYHWD. Compared with the model group, BYHWD and the optimized one could evidently reduce cerebral cortex infarction area and improve the impaired brain edema (P<0.05), and the medium-dose group was the best. Conclusion The optimized BYHWD ratio is:Astragali Radix:Chuanxiong Rhizoma:Pheretima=10:3:1.

Objective To investigate influence of combined use of allopurinol and warfarin on the INR of chronic permanent atrial fibrillation hyperuricemia patients. Methods 80 cases of patients with chronic atrial fibrillation complicated with hyperuricemia, in Sanmen County People's Hospital from February 2016 to February 2017 were randomly divided into the control group (n=40) and the observation group (n=40), the control group was given warfarin plus low purine diet treatment, the observation group was given additional Allopurinol treatment. Changes of INR were compared. Results There was no statistical significance in INR level before treatment between the two groups of patients with permanent atrial fibrillation complicated with chronic hyperuricemia. The INR level of the control group was not changed after treatment, the observation group increased significantly after treatment, and there was statistical significance between the two groups (P<0.05). There were no bleeding events in the control group, there were 6 cases of the observation group with minor bleeding and subcutaneous ecchymosis, including 2 cases with fecal occult blood, 1 cases with hematuria, nasal hemorrhage . The change of liver and kidney function was not obvious. Conclusion In application of allopurinol treatment plus warfarin for patients with chronic persistent atrial fibrillation and hyperuricemia patients, , INR need to be regularly monitored, in order to provide reference for Warfarin dosage adjustment, and ensure clinical safety.

The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

OBJECTIVE: To explore alcohol pharmacokinetics as well as acetaldehyde level in peripheral blood in human subjects with different ALDH2 genotypes after drinking. METHODS: Venous blood samples of 14 unrelated volunteers were collected. Polymerase chain reaction-restriction fragment length polymorphism technology was adopted for DNA extraction and ALDH2 genotyping. The volunteers were asked to drink beer at certain doses. The concentration of alcohol and acetaldehyde were assayed by headspace gas chromatography method at different time. The pharmacokinetic parameters were calculated. RESULTS: According to the results of electrophoresis, 5 people carried ALDH2*1/*1 as wild group and 9 people carried ALDH2*1/*2 as mutation group. The good linear range of alcohol and acetaldehyde were 0-1 570.7 microg/mL and 0-5.1772 microg/mL, respectively. The AUC values of alcohol and acetaldehyde and the t1/2Z value of alcohol were higher in the mutation group than that in the wild group. But the CL/F value of alcohol was lower in the mutation group than that in the wild group (P<0.05). CONCLUSION After the consumption of alcohol, alcohol and acetaldehyde metabolism in blood slow down in ALDH2*1/*2 mutation group influenced by the inhibition of enzyme activity, leading to the accumulation of acetaldehyde in peripheral blood, thus reinforcing their effects in the body.
Alcohol Drinking ; Aldehyde Dehydrogenase/genetics* ; Aldehyde Dehydrogenase, Mitochondrial ; Ethanol/metabolism* ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic

BACKGROUNDIt has been reported that there is a significant difference in the local tissue concentration of transforming growth factor (TGF)-β1 between chronic rhinosinusitis without nasal polyps (CRSsNP) and chronic rhinosinusitis without nasal polyps (CRSwNP) patients. TGF-β has been reported to play an important role in regulating epithelial cell repair in lower airway remodeling and may be a critical factor involved in the remodeling process of chronic rhinosinusitis (CRS).

METHODSEthmoidal mucosal samples collected from CRS and healthy control patients were analyzed for TGF-β1, TGF-β receptor I, TGF-β receptor II, Smad3, phospho-Smad3, Smad7, and Smad anchor for receptor activation by Western blotting analysis. The proliferation of sinonasal epithelial cells at baseline and after TGF-β1 and/or EGF stimulation was evaluated by the MTT assay.

RESULTSIn CRSsNP, TGF-β1, TGF-β receptor I, TGF-β receptor II, and Smad3 protein levels were significantly higher than controls. In CRSwNP, TGF-β1, Smad3, and pSmad3 protein levels were significantly lower than controls. Smad7 protein was significantly higher in CRS than controls. In vitro experiments demonstrated that the baseline proliferation levels of sinonasal epithelial cells were lower in CRS than controls.

CONCLUSIONSCRSwNP is characterized by a lower level of TGF-signaling compared with the control. In CRSsNP, although the upstream signaling of TGF-β was enhanced, the high Smad7 protein expression may restrain the downstream signaling components (e.g., pSmad3) and the TGF-β antiproliferative effect on sinonasal epithelium. The difference in the local tissue concentration of TGF-β1 between CRSsNP and CRSwNP patients did not result in significant differences in epithelial proliferation.

Adult ; Aged ; Benzamides ; pharmacology ; Cells, Cultured ; Dioxoles ; pharmacology ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; Receptors, Transforming Growth Factor beta ; antagonists & inhibitors ; metabolism ; Serine Endopeptidases ; metabolism ; Signal Transduction ; drug effects ; Sinusitis ; metabolism ; Smad3 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; antagonists & inhibitors ; metabolism ; Young Adult

Objective To investigate the correlation between the gene polymorphism of homocysteine metabolic enzyme cystathionine β-synthase(CBS) 844ins68,N5,10-methylenetetrahydrofolate reductase(MTHFR) C677T and chronic pulmonary heart disease(CPHD).Methods A total of 230 patients with CPHD in observation group were selected from January 2014 to November 2016 in the Second People's Hospital of Xinxiang City,and 235 healthy subjects in healthy control group were selected at the same time.The lung function test was performed with lung function instrument,and the percentage of the forced expiratory volume in one second to predicted value(FEV1％ pred) and the forced expiratory volume in one second to forced vital capacity(FEV1％) value were recorded in the two groups.The fasting ulnar venous blood was collected from the patients in the observation group on the next morning after hospitalization and the subjects in the control group on the morning of health examination.The levels of plasma homocysteine (Hcy),fasting blood glucose (FBG),triacylglycerol (TG),total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were detected.The DNA was extracted from the whole blood cells.The CBS 844ins68 polymorphism was detected by polymerase chain reaction genotyping.The MTHFR C677T polymorphism was detected by restriction fragment length polymorphism polynerase chain reaction.Results There was no significant difference in the FBG level between the two groups (P ＞ 0.05).The levels of Hcy,TG,TC and LDL-C in the observation group were significantly higher than those in the healthy control group (P ＜ 0.05),and the FEV1 and FEV1％ pred were significantly lower than those in the healthy control group (P ＜ 0.05).There were two genotypes of CBS 844ins68 in the two groups.The genotype frequencie of DD and DI in the observation group was 91.74％ and 8.26％,and the allele frequency of D and I was 95.87％ and 4.13％ respectively.The genotype frequency of DD and DI in the healthy control group was 94.04％ and 5.96％,and the allele frequency of D and I was 97.02％ and 2.98％ respectively.There was no significant difference in genotype and allele frequency distribution between the two groups (x2 =0.935,0.901;P ＞ 0.05).Three genotypes of MTHFRC677T were detected after enzyme digestion in the two groups.The genotype frequency of CC,CT and TT in the healthy control group was 27.66％,48.94％ and 23.40％;and the allele frequency of C and T was 52.13％ and 47.87％ respectively.The frequency of TT genotype and T allele in the observation group was significantly higher than that in the healthy control group (x2 =7.730,7.326;P ＜ 0.05).Conclusions Hcy level increasing may be a risk factor for CPHD.The polymorphisms of CBS 844ins68 gene may be unrelated to the occurrence of CPHD.The polymorphism of the MTHFR C677T gene may contribute to CPHD by affecting Hcy level.The T allele of MTHFR C677T may be a risk factor for CPHD,and the MTHFRC677T gene may be a genetic predisposition to CPHD.