Humans ; Protein Kinase Inhibitors ; therapeutic use ; Protein-Tyrosine Kinases ; antagonists & inhibitors

Objective: To establish a method to determine 4 active ingredients in Jindan tablets by HPLC.
Methods: Agilent Eclipse X DB C₁₈ (4.6 mm×250 mm, 5 μm) column was adoped using acetonitrile (phase A) -0.1% phophoric acid solution (phase B) as the mobile phase with gradient program at the flow rate of 1.0 mL·min^-1, the detection wavelength was 270 nm, Column temperature was set at 30 ℃.
Results: The linear ranges of gentiopicrin, polydatin, quercetin and emodin were 7.875-78.75 μg·mL^-1（r=0.999 9）, 6.75-67.50 μg·mL^-1（r=0.999 7）, 7.726-77.26 μg·mL^-1（r=0.999 4）, 3.809-38.09 μg·mL^-1（r=0.999 8), respectively, the peak area had a good linear relationship with the mass concentration of 4 components, the average recovery were 99.31%, 99.21%, 99.04%, 99.59%, (n=6) respectively, and RSDs were 1.86%, 1.24%, 1.37%, 1.15%, respectively.
Conclusion: This method is reliable, repeatable and stable, and has strong specificity. This method can provide a reference for the quality control and standard improvement of Jindan tablets.

Objective To elucidate the prevalence and clinical characteristics of human metapneumovirus(hMPV)in hospital- ized children with respiratory infection.Methods A total of 452 hospitalized children with lower respiratory tract infection were observed from Aug 2004 to Jan 2005.Respiratory tract aspirates were collected from all patients within 48 hours after admis sion.The specimens were routinely tested for respiratory syncytial virus,influenza virus A and B,parainfluenza virus 1 to 3 and adenovirus by direct fluorescent assay(DFA).The 245 specimens negative by DFA were tested for hMPV by RT-PCR. PCR products of hMPV M gene from some patients were randomly selected for sequencing analysis.Results hMPV was identi- fied in 59(24.1%)of the 245 specimens tested,hMPV infection alone accounted for 13.1% of the infections in the 452 chil- dren under study,The prevalence of hMPV was higher than other respiratory viruses in winter.The mean age of hMPV-infec- ted children(n=59)was 27.7 months.There was no significant difference between age groups in terms of the prevalence of hMPV(P＞0.05).There were no statistically significant difference in demographics and clinical symptoms between hMPV in- fection and other common respiratory virus infection.Genotyping for the hMPV M gene from 23 Shanghai patients showed two distinct hMPV genotypes.Sequence analysis of these hMPV M genes showed 82.8%-100% homology to the registered se- quence in GenBank.There was no significant difference in clinical characteristics between the 2 genotypes.Conclusions hMPV plays an important pathogenic role in lower respiratory tract infection of children,hMPV prevailed in the winter of 2004.Clini- cally,hMPV infection can not be discriminated from the infection of other respiratory viruses.Clinical manifestation is similar between the two hMPV genotypes.

Objective To observe the effect of bilateral movement training on upper extremities dysfunction in stroke patients in convalescent phase. Methods 52 patients with hemiplegia were randomly divided into treatment group (n=26) and control group (n=26). The treatment group accepted bilateral upper extremities movement training, and the control group accepted routine neurodevelopment training mainly with affected upper extremities, for 6 weeks. They were assessed with Fugl-Meyer Assessment upper-extremity section (FMA-UE) and modified Barthel Index (MBI) before and after treatment. Results The FMA-UE and MBI scores improved in both groups after treatment (P<0.01), and improved more in the treatment group than in the control group (P<0.001). Conclusion Bilateral movement training may improve upper extremity function and activities of daily living more effectively for stroke patients in convalescent phase.

Objective To investigate the effects of zinc deficiency on the expressions of Th1/Th2 cytokine interferon-?(IFN-?) and interleukin-4(IL-4)in asthmatic rats.Methods Animal models of asthma and zinc deficiency were established.Thirty-two SD rats were divided into 4 groups according to weight:zinc deficient diet with ovalbumin(OVA) challenge group(group A),zinc normal diet with OVA challenge group(group B),zinc normal pair-fed diet with OVA challenge group(group C) and zinc normal diet with saline challenge group(group D).The contents of IFN-? and IL-4 in lung homogenate were detected by enzyme linked immunosorbent assay(ELISA).The levels of IFN-? and IL-4 mRNAs were observed by reverse transcriptase-polymerase chain reaction(RT-PCR).Results Compared with group D,the content and mRNA expression of IFN-? of lung in group A,group B and group C all decreased significantly(all P0.05).No marked difference was found between group B and group C in all the results.Conclusions Zinc deficiency reduces the expression of IFN-? but have no influence on IL-4.The imbalance of Th1/Th2 cytokine may be associated with the increasing airway inflammatory reaction in zinc deficient asthmatic rats.

Objective To study the expression level of thyroid insulin-like growth factor-Ⅰ (IGF-Ⅰ) in iodine deficiency and excess mice and the effect of thyroid gland IGF-Ⅰ in the thyroid morphological change. Methods Forty-eight Balb/c mice were chosen as studied objects,weighing about 16 g. They were divided into three groups: low iodine(LI,iodine content of 50 μg/kg in feed,drinking deironized water) group,normoi(NI,iodine content of 300 μg/kg in feed,drinking deironized water) group and high(HI,iodine content of 300 μg/kg in feed,iodoine of content 14 700 μg/kg in drinking) group,16 mice in each group. Mice were put to death after 12 weeks and taken out of their thyroid gland. The body weight,absolute and relative weights of thyroid gland were measured and the morphological change of thyroid gland were observed under microscope. The expression levels of thyroid gland IGF-Ⅰ mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively. Results There were statistical significances between groups of thyroid absolute and relative weights(F = 315.881,405.921,all P < 0.01). LI group [(10.71±4.03) mg,(44.98±15.39)mg/100 g body weight]and HI group [(3.42±1.17)mg,(13.50± 3.89)mg/100 g body weight]had heavier thyroid absolute and relative weights than NI group[(2.11±0.53)mg,(8.35±1.98)mg/100 g body weight,all P < 0.01]. Under microscopy,the thyroid follicle capacity grew down and the follicle quantity grew up in LI group,the epithelium was stylolitic,the colloid diminished or absence in follicular cavity,while HI group presented colloid accumulation without follicular hyperplasia. The expression level of thyroid gland IGF-Ⅰ mRNA in LI group(1.03±0.32) was more than that in NI(0.65±0.19) and HI(0.59± 0.20) groups(F= 7.518,P< 0.01). In contrast to NI,there was no difference in the expression level of thyroid gland IGF-Ⅰ mRNA in HI group(P > 0.05). The brownish particles of LI group were more than NI and HI groups in the thyroid follicle epithelium by immunohistochemistry,while HI group was less than NI group. Conclusions The mice of iodine deficiency presented follicular hyperplasia goiter,the mice of iodine excess presented colloid accumulative goiter. The change of IGF-Ⅰ mRNA and protein expression may participate morphologleal change,indicating autocrine IGF-Ⅰ of thyroid gland may play an important role in regulating goiter formation.

Objective:To investigate the electrocardiographic characteristics of left and right ventricles origin of premature ventricular contractions(PVCs) during V3 transition of precordial leads, right ventricular outflow tract (RVOT) anterior septum and right coronary sinus (RCC), and RVOT middle-posterior septum and left coronary sinus (LCC).Methods:From January 2017 to September 2019, 91 patients with ventricular extrasystole of outflow tract who had V3 transition in precordial lead and had successful radiofrequency ablation in RVOT anterior septum, middle posterior septum, LCC and RCC were selected for retrospective case control study.The electrocardiography measurements of PVCs were compared between the anteroseptal RVOT group and RCC group, as well as the middle-posterior septal RVOT group and the LCC group, respectively.The measurements included the R-wave amplitude in lead Ⅰ, Ⅱ, Ⅲ and aVF, R amplitude ratio in leads Ⅲ to Ⅱ, Q-wave amplitude in lead aVL and aVR, Q amplitude ratio in leads aVL to aVR, R-wave and S-wave amplitude from leads V1 to V3, the V2S/V3R index, the transition zone index, and the V2 transition ratio.Results:Thirty-six cases originated from the anteroseptal RVOT, and 11 from the LCC.Lead I R-wave amplitude in anterior septal RVOT was higher than LCC group((0.22±0.25) mV vs.(-0.17±0.33) mV; P=0.003). R-wave amplitude in lead Ⅱ was lower than that in the LCC group((1.59±0.35) mV vs.(1.76±0.27) mV; P=0.035). R-wave amplitude in lead aVF was lower compared with the LCC group((1.53±0.35) mV vs.(1.78±0.39) mV; P=0.050). The V2S/V3R index showed a significant difference between these two groups(1.99±0.66 vs.0.76±0.38; P<0.001). The V2 transition ratio also appeared a significant difference between the two groups(0.69±0.43 vs.1.05±0.35; P=0.005). PVCs arose from the middle-posterior septal RVOT in 32 cases, and from the RCC in 12 cases.Compared with RCC group, lead Ⅰ R-wave amplitude showed lower ((0.25±0.31) mV vs.(0.57±0.12) mV; P<0.001); R amplitude ratio in leads Ⅲ to Ⅱ higher (0.89±0.14 vs.0.72±0.18; P=0.002); Q amplitude in lead aVL((0.72±0.24) mV vs.(0.51±0.16) mV; P=0.002)higher, and Q amplitude ratio in leads aVL to aVR higher in the middle-posterior septal RVOT(0.76±0.23 vs.0.50±0.21; P=0.002). Conclusion:Among the cases with lead V3 transition, PVCs originated from the anteroseptal RVOT show significantly different R wave in lead Ⅰ, Ⅱ, aVF, V2S/V3R index, and the V2 transition ratio compared with those from the LCC.The PVCs from the middle-posterior septal RVOT and the RCC have different R wave in lead Ⅰ, R amplitude ratio in leads Ⅱ and Ⅲ, Q amplitude ratio in leads aVL and aVR.Combined with its different characteristics, it can help to identify the origin of left and right ventricles.

OBJECTIVETo explore the intervention of baicalin on signal transduction and activating transcription factor expression of ulcerative colitis (UC) patients.

METHODSRecruited were UC patients at Outpatient Department of Digestive Disease, Inpatient Department of Digestive Disease, Center for Digestive Endoscopy of College City Branch, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Southern Hospital affiliated to Southern Medical University from June 2010 to January 2011. They were assigned to the UC group (33 cases) and the diarrhea-predominant irritable bowel syndrome (IBS-D) group (30 cases). Another 30 healthy subjects were recruited as a healthy control group. Peripheral blood mononuclear cells (PBMCs) in vitro intervened by different concentrations baicalin were taken from UC patients. IL23R gene expressions in vitro intervened by different concentrations baicalin were detected using Q-PCR. Expressions of signal transducer and activator of transcription 4 (STAT4) , STAT6, phosphorylated-STAT4 (p-STAT4), and p-STAT6 were detected using Western blot. Serum levels of IFN-γ, IL-4, IL-6, and IL-10 were measured by ELISA. Effects of different concentrations baicalin on expressions of PBMCs, and levels of IFN-γ, IL-4, IL-10 of UC patients were also detected.

RESULTSCompared with the negative control group, 40 µmol baicalin obviously decreased IL23R gene expression of UC patients (P <0. 01). Compared with the healthy control group and the IBS-D group, p-STAT4/STAT4 ratios increased, p-STAT6/STAT6 ratios decreased, levels of IFN-γ, IL-4, IL-10 all increased in the US group (all P <0. 05). Compared with the negative control, 5 and 10 µmol baicalin groups, 20 and 40 moL baicalin obviously decreased p-STAT4/STAT4 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously increased p-STAT6/STAT6 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously lowered levels of IFN-γ and IL-4, and elevated IL-10 levels (all P <0. 05).

CONCLUSION40 µmoL baicalin could in vitro inhibit p-STAT4/STAT4 ratios, adjust p-STAT6/STAT6 ratios and related cytokines, thereby balancing the immunity and relieving inflammatory reactions of UC.

Activating Transcription Factors ; metabolism ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Blotting, Western ; Colitis, Ulcerative ; drug therapy ; metabolism ; Cytokines ; metabolism ; Flavonoids ; therapeutic use ; Humans ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; Leukocytes, Mononuclear ; Medicine, Chinese Traditional ; Phosphorylation ; STAT6 Transcription Factor ; metabolism ; Signal Transduction

OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.

METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.

RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.

CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.

Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic

BACKGROUNDProteins or peptides can be directly transferred into cells when covalently linked to protein transduction domains (PTDs). TAT is one of the most widely studied PTDs. The effect of fusion protein TAT and heme oxygenase-1 (HO-1) on liver sinusoidal endothelial cells (SECs) apoptosis during cold storage is unknown. The present study aimed to determine whether fusion protein TAT-HO-1 would transduce efficiently into liver during cold storage, and, if so, to determine whether TAT-HO-1 would attenuate SECs apoptosis during preservation injury in rat.

METHODSLivers of Sprague-Dawley rats were harvested and randomly assigned to group 1 (HTK solution) and group 2 (HTK solution containing TAT-HO-1 fusion protein) according to the type of the preservation solution. The transduction efficiency of TAT-HO-1 was examined and the impairment of SECs was assessed during the period of cold storage followed by 1 hour of reperfusion.

RESULTSTAT-HO-1 can transduce efficiently into liver during cold storage. A significantly lower apoptotic index of SECs was observed in group 2, at 6, 12 and 18 hours of cold storage after 1 hour reperfusion, when compared with group 1. TAT-HO-1 reduced HA and ET levels in liver at each time point. Both Bcl-2 and Bax protein were expressed in hepatocytes and SECs at the periphery of the sinusoidal space. Moreover, higher Bcl-2 expression and lower Bax expression were observed in group 2.

CONCLUSIONSTAT-HO-1 can transduce efficiently into rat livers and shows a protective effect on SECs by attenuating apoptosis during cold ischemia/reperfusion injury. Protein transduction will be a novel therapeutic strategy to reduce the risk of preservation injury in liver transplantation.

Animals ; Apoptosis ; drug effects ; Endothelial Cells ; cytology ; drug effects ; Heme Oxygenase-1 ; genetics ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Liver ; cytology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; bcl-2-Associated X Protein ; metabolism ; tat Gene Products, Human Immunodeficiency Virus ; genetics