Objective:To establish a method of multi-PCR to amplify the complete DNA sequence (CDS) of TCR ? and ? chain of the antigen-specific T lymphocytes in local pathologic specimen of active pulmonary tuberculosis patients, and to analyze ?/? T cell receptor gene rearrangement and CDR3 repertoire.Methods:The lymphocytes in bronchoalveolar lavage (BAL) of active pulmonary tuberculosis patients were separated. Following total RNA extraction, cDNA synthesis, Multi-PCR, recombinant clones construction, and sequencing, the CDS of TCR ? and ? chains from these lymphocytes were analyzed by using software of DNAstar and internet TCR resources.Results:24 of ? chain CDS and 13 of ? chain CDS from 3 samples of BAL were obtained. As for TCR ? chain, AV1S2 (54%), AV12S3 (41%), and AV12S2(5%) appeared frequently. BV2(38%), BV29S1(46%), BV14(3%), and BV4S2(3%) in TCR ? chain appeared more often. There were CDR3 diversities between samples and even in the same sample by amino acid sequence analysis, but there were a few identical or similar amino acid sequences. There was the same amino acid sequence of SVGTGTLHQETQY in CDR3 region of ? chain of BAL sample No.1 and No.2; The sequence of AVRDWAGNMLT appeared in two ? chains of BAL sample No.2 and No.3; Moreover, the sequence of AV…DNN…RLM appeared in ? chains of BAL sample No.2 and No.3.Conclusion:A method of Multi-PCR is used to amplify TCR ? and ? chain CDS of tuberculosis patients. There are characteristic T cell clones to proliferate,with TCR ? and ? chain repertiore skewing in local infective focus. The sequences of CDR3 in different TCR clones are mostly different but there are a few identical or similar sequences in the same patient or even between different patients. The identical amino acid sequences of CDR3 are possibly specific for recognizing MTB polypeptide.

Objective To evaluate the utility of CD4+ TCR tetramers‐based flow cytometric analysis and cell climbing slice assay in detecting antigen‐specific CD14+ monocytes in the blood of tuberculosis (TB) patients .Methods CD4+ TCR tetramers were used to detect tetramer‐positive CD14+ monocytes in the peripheral blood (PBL ) samples of inpatients with advanced pulmonary TB (PTB) by flow cytometric analysis .The PBL samples obtained from non‐TB patients and umbilical cords were used as controls .These tetramers were also used to examine tetramer‐bound CD14+ monocytes and Mycobacterium tuberculosis (MTB) antigen‐specific and tetramer‐bound cells by cell climbing slice in situ staining .Results The median percentage of tetramer‐bound CD14+ monocytes in PBL samples from PTB patients ,non‐TB patients and umbilical cords were 1 .32% , 0 .50% and 0 .26% respectively by using CD4+ Vα21‐J39/Vβ29‐D1‐J2 tetramer , while the medians were 1 .05% , 0 .49% and 0 .19% respectively by using CD4+ Vα21‐J39/Vβ29‐D2‐J2 tetramer . The percentage of tetramer‐bound CD14+ monocytes in PTB patients group was significantly higher than the other two control groups .In cell climbing slice in situ staining ,tetramer‐bound CD14+ monocytes ,and MTB antigen‐specific and tetramer‐bound cells were positive in PTB tissue compared with negative in control tissues . Conclusions CD4+ TCR tetramers‐based flow cytometric analysis and cell climbing slice assay could be used to sensitively detect M TB antigen‐specific CD14+ monocytes in the blood of TB patients ,and more accurately evaluate the changing profile and clinical significance of these cells in TB patients .

OBJECTIVE:To improve the quality standard of Shuangshen capsule. METHODS:TLC was conducted for the qual-itative identification of Curcumnae radix and Rehmannia glutinosa in the preparation. HPLC was used for the content determination of salvianolic acid B in the preparation:the column was Diamonsil C18(2)with mobile phase of acetonitrile-0.1% H3PO4(22:78, V/V) at a flow rate of 0.5 ml/min,detection wavelength was 286 nm,column temperature was 30 ℃,and the injection volume was 2 μl. RESULTS:The TLC spots of C. radix and R. glutinosa were clear and well separated;the linear range of salvianolic ac-id B was 0.0792-0.792 μg(r=0.9999);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 100.71%-101.82%(RSD=0.50%,n=6). CONCLUSIONS:The established standard can be used for the quality control of Shuang-shen capsule.

0.05).Conclusions The chemotherapeutic embedding radioactive particle 125Ⅰ combined with GP is safe,feasible,and effective to the therapy of senile-inoperable NSCLC.

Objective : To learn the characteristics of second-line drug resistance and related gene mutations of multidrug-resistant Mycobacterium tuberculosis ( MDR-TB ) Beijing genotype strains. Methods: The MDR-TB isolates in Hwa Mei Hospital from 2017 to 2019 were enrolled and detected using RD105 deletion-targeted multiplex polymerase chain reaction (PCR). The proportion method for drug susceptibility test was used to detect the drug-resistant profiles against kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin. The gene sequencing of rrs, tlyA, eis, gidB, gyrA and gyrB was conducted by PCR compared with H37RV strain. The differences in the rates of drug resistance and mutation between Beijing and non-Beijing genotype strains were examined to understand the characteristics of Beijing genotype strains. Results: There were 106 Beijing genotype and 27 non-Beijing genotype strains in 133 MDR-TB isolates. The drug resistance rates of kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin in Beijing genotype strains were 9.43%, 7.55%, 3.77%, 32.08% and 32.08%, respectively. The rates of quasi-extensive and extensive drug resistance in Beijing genotype strains were 30.19% and 7.55%. The gene mutation rates of rrs, tlyA, eis, gidB, gyrA and gyrB in Beijing genotype strains were 7.55%, 7.55%, 1.89%, 2.83%, 36.79% and 2.83%, respectively. There were no significantly differences between Beijing and Non-Beijing genotype strains in the factors above ( P>0.05 ). The gene rrs, tlyA, eis, gidB, gyrA and gyrB had 2, 1, 2, 2, 5 and 3 mutation types, respectively, with single base substitution as the main type. Conclusion Beijing genotype strains are dominant in MDR-TB, with high resistance to fluoroquinolones and mainly gyrA gene mutation.

Objective To evaluate ketamine-induced cerebral protection in mice with traumatic brain injury (TBI) by magnetic resonance imaging (MRI).Methods Thirty-two pathogen-free healthy male C57BL/6 mice,aged 8 weeks,weighing 26-30 g,were divided into 4 groups using a random number table:control group (group C,n=7),ketanine group (group K,n=7),TBI group (n=9) and TBI plus ketamine group (group TBI+K,n =9).TBI was produced with a pneumatically driven controlled cortical impact device.Ketamine 150 mg/kg was intraperitoneally injected at l h after operation in TBI+K and K groups,while the equal volume of normal saline was given instead in TBI and C groups.Open field test was conducted at 24 h,72 h and 7 days after operation.The animals in TBI and TBI+K groups were scanned by T1-weighted MRI at 6,24 and 72 h after operation,the animals in C and K groups were scanned by MRI at 24 h after operation,and the development of cerebral edema was observed.Results MRI scan showed no cerebral edema in C and K groups,and different degrees of cerebral edema were found in TBI and TBI+K groups.Compared with group C,the locomotor distance was significantly shortened at 24 and 72 h after operation in group TBI (P＜0.05).Compared with group TBI,the size of cerebral edema was significantly decreased,and the locomotor distance was prolonged at 24 and 72 h after operation in group TBI+K (P＜0.05 or 0.01).Conclusion MRI method further clarifies that ketamine can produce cerebral protection to some extent in mice with TBI.

Objective To explore the effect of IL-1? and IL-6 on MMP-3 gene expression in human coronary artery smooth muscle cells.Methods We used IL-1?(20 ?g/L) and IL-6(10 ?g/L) to stimulate human coronary artery smooth muscle cells,which were co-culture for 0,2,4,8,24,36 h.IL-1?(0,5,20,40 ?g/L) and IL-6(0,5,10,50 ?g/L) were used to stimulate human coronary artery smooth muscle cells,which were co-cultured for 6 h.Then we detected the gene expression by fluorescent quantitation PCR.Results In the same concentration of IL-1? and IL-6,gene expression was up-regulated at 2 h,at 8 h the expression reached the peak,then began to descend.In different concentration of IL-1? and IL-6,gene expression was up-regulated with the dose of IL-1? and IL-6(IL-1?: r=0.907,P=0.000;IL-6: r=0.919,P=0.000).There were significant differences in MMP-3 expression among different groups(IL-1?: F=24.047,P=0.000;IL-6: F=14.081,P=0.001).There were no significant differences in matrix metalloproteinase-3 between IL-1? 20 and 40 ?g/L groups(P=0.154) and between IL-6 5 ?g/L and 10 ?g/L(P=0.292).Conclusion It suggests that IL-1? and IL-6 can promote MMP-3 gene expression in human coronary artery smooth muscle cells,and it may be one of the mechanisms of inflammation effect in acute coronary syndrome.

AIM:To investigate the effect of cervical lymphatic blockage(CLB)on blood pressure(BP)in conscious unrestrained rats.METHODS:Sprague-Dawley(SD)rats were adopted and randomly divided into two groups as Sham operated group and CLB group.By means of monitoring hemodynamic change in conscious unrestrained rats,twenty-four-hour blood pressure(SBP,DBP and MAP),blood pressure variability(BPV),heart rate(HR)and heart rate variability(HRV)were respectively measured before sham and cervical lymphatic blockage operation as their baseline and at 1st,3rd,7th,11th,15th days after operation.Meanwhile,arterial baroreflex sensitivity(BRS)was measured before and at 1st,7th,15th days after operation.RESULTS:SBP,DBP,MAP and HR significantly decreased at 1st day after CLB operation and their lowest values appeared at 7th day.The tendency of their alternation was descending early and then ascending whereas reverse alterations of BPV and HRV were observed.BRS reduced in CLB rats with no apparent recovery from 7th day.CONCLUSION:CLB results in reduction of blood pressure and dysfunction of nervous regulation on cardiovascular system in conscious unrestrained rats.

OBJECTIVE: To study the characteristics of adverse drug reactions( ADR) in our hospital. METHODS: 240 ADR cases collected between 2003 and 2006 in our hospital were analyzed retrospectively. RESULTS: Of the total ADR cases, 211( 87. 92% ) were induced by intravenous drugs, 153( 63. 75% ) were induced by antibacterials, and 118( 49. 17% ) were manifested as lesions of skin and its appendages. CONCLUSIONS: Importance should be attached to the monitor and reporting of ADR so as to lessen or avoid the occurrence of ADR.

Aim To investigate the vasodilatory effect of Ferulic acid on in vitro rat coronary artery and its possible mechanism. Methods By using the mi-crovessel tension recorder system, the vasodilatory effect of FA on resting and contractin-vitro rat coronary artery was determined;the influence of endothelial in-tegrity to FA-induced vasorelaxation was observed; the relationship of FA on [ Ca2+] ex-influx-induced and [ Ca2+] in-efflux-induced contractions was discussed;the mechanism of vasodilatory effect of FA was ex-plored by applying the inhibitors of KCa(TEA),KATP channel ( Gli ) , KIR channel ( BaCl2 ) , KV ( 4-AP ) , NOS( L-NAME) and COX( Indo) . Results FA had no effect on the resting tension of in vitro rat coronary artery. FA dilated the in-vitro rat coronary artery pre-treated with KCl ( 60 mmol · L-1 ) , U46619 ( 1 μmol · L-1 ) and PE ( 10μmol · L-1 ) in a concentration-dependent fashion ( P < 0. 05 ) . FA inhibited the [ Ca2+] ex-influx-induced and [ Ca2+] in-efflux-in-duced contractions significantly ( P <0. 05 ) . 4-AP ( 1 mmol· L-1 ) restrained the diastolic function of FA, while TEA, Gli, BaCl2、L-NAME, Indo had no obvious effect ( P >0. 05 ) . Conclusion The diastolic func-tion could be related to the activation of KV channel on vascular smooth muscle cells, the free Ca2+ from Sar-coplasmic reticulum cells and blockade extracellular Calcium channel do not depend on KCa, KATP, KIR channel, nor the endothelial function.