Purpose:To study the effect of the herbal decoction Qingyi Xiaoji Formula(QYXJ) on the proliferation of human pancreatic cancer cell line SW1990 in vivo and to explore the mechanism of its functions by means of cDNA microarray.Methods:Tumor-burdened nude mice were randomized into control group,5-FU group,and QYXJ groups at different dosages.After treatment,inhibiting rates of tumor growth were calculated.Tumor mRNA of the control group and the QYXJ group at moderate dose was extracted.The fluorescent cDNA probes were prepared,labeled with two different dyes Cy3 and Cy5,and then hybridized with cDNA microarray and scanned for fluorescent intensity.The genes with different expression were identified through the analysis of gene expression profile.Results:Inhibition rates of tumor growth in the QYXJ groups were 21.31%,38.16% and 29.09%,in the dose of 18 g/kg,36 g/kg,and 72 g/kg respectively.7 genes with reduced expression were identified,the functions of which were oncogene,protein translation and synthesis,DNA synthesis and repair,cell signal transduction,etc.Conclusions:QYXJ decoction may inhibit the proliferation of pancreatic cancer in vivo,possibly by blocking the action of an oncogene and its downstream signaling,or by regulation of protein synthesis in cancer cells.

Objective To investigate the drug resistant genes and genotypes of carbapenem-resistant Klebsiella pneumoniae in Tianjin First Center Hospital. Methods A total of 52 strains of carbapenem-non-susceptible Klebsiella pneumoniae were collected from 2012 to 2015. The MICs of antimicrobial drugs were detected using agar dilution methods. Phenotypes of carbapenemases were screened using modified Hodge test. Drug resistant genes were detected by multiplex-PCR assay. Multilocus sequence typing ( MLST) was used to determine the genotypes and homology of these carbapenem-resistant Klebsiella pneumoniae strains. Results Susceptibility of antimicrobial agents indicated that all these strains with multiple drug resistance. The resistance rate to piperacillin/tazobactam, ceftriaxone, ceftazidime, cefepime, aztreonam imipenem,meropenem was 100%( 52/52 ) . The resistance rate of ST11 type to amikacin was 93. 5%( 43/46), ciprofloxacin was 97. 8%(45/46), levofloxacin was 97. 8%(45/46), compound sulfamethoxazole was 17. 4%(8/46), tigecycline was 0. The resistance rate of ST101 type to amikacin was 3/3, ciprofloxacin was 2/3, levofloxacin was 3/3, compound sulfamethoxazole was 3/3, tigecycline was 0. The resistance rate of ST709 type to amikacin was 1/1, ciprofloxacin was 1/1, levofloxacin was 1/1, compound sulfamethoxazole was 1/1, tigecycline was 0. The resistance rate of ST1393 type to amikacin was 1/1, ciprofloxacin was 1/1, levofloxacin was 1/1, compound sulfamethoxazole was 1/1, tigecycline was 0. The resistance rate of ST2068 type to amikacin was 1/1, ciprofloxacin was 1/1, levofloxacin was 1/1, compound sulfamethoxazole was 1/1, tigecycline was 0. PCR results showed that 43 isolates were blaKPC-2 positive and 5 isolates were blaOXA-48 positive, 1 isolate was blaDNM-1 positive. There were 46 strains of ST11 type. The 43 strains of Klebsiella pneumoniae producing KPC-2 type carbapenemase were all ST11. While among 5 strains of Klebsiella pneumoniae carrying OXA-48 carbapenem resistant gene, 3 strains were ST101, 1 was ST709, 1 was ST1393. One strain of Klebsiella pneumoniae harboring DNM-1 type carbapenemase was ST2068. Conclusions Drug resistant genes of carbapenem-resistant Klebsiella pneumoniae were KPC-2 dominant, OXA-48 and DNM-1 were sporadical;the genotype was mainly ST11 by MLST in the hospital. The research provided effective basic and reference for the hospital infection t control.

Objective To summarize and analyze regularities in clinical use of acupoint application for headache. Method Fifth version Chinese Medical Classics database was searched. Thirty-three library-stored ancient books were searched manually. The ancient literature included for the study was sorted out. Descriptive statistical analysis was made of classification, point selection, used medicine, excipients and dosage forms for acupoint application for headache. Result A total of 51 pieces of literature was included. Statistical analysis showed that there were nine kinds of disease names, three types of classification-based treatment, eight acupoints, fifty kinds of medicines, eight hinds of excipients and five kinds of dosage forms. Conclusion Headache is the most basic nomenclature for pains in the head. Classification-based treatment fully embodies the idea of treatment based on disease differentiation mainly in combination with syndrome differentiation. Point Taiyang is the main point for local selection of point. The most frequently selected medicines are those of going through meridians, opening the orifices, unblocking collaterals and having thick smells. Blistering medicines are used cautiously. Scallion juice is most frequently used as excipients. Medicinal cakes are dosage forms most suitable for acupoint application for headache.

Objective To quantitatively determine JAK2V617F mutation by fluorescence quantitative polymerase chain reaction (FQ-PCR)and to analyze its diagnostic value in chronic myeloproliferative disorders(CMPD).Methods FQ-PCR was adopted to detect the positive rate and the relative quantification of JAK2V617F gene mutation in the marrow samples of 68 patients with CMPD.Their diagnostic value in CMPD and their relation with the clinical data were analyzed.Results The marrow samples in 68 cases of CMPD were successfully amplified in wild-type and mutant JAK2V617F.The total positive rate of the JAK2V617F gene mutation was 66.2%(45/68).The positive rate was 75.0%(21/28)in the patients with polycythemia vera(PV),60.6%(20/33)in the patients with primary thrombocythemia(ET)and 57.1%(4/7)in patients with idiopathic myelofibrosis respectively.The posi-tive rate of JAK2V617F mutation in the PV patients was significantly higher than that in the patients with ET and IMF.The JAK2V617F relative quantification in the PV patients was positively correlated with the hemoglobin concentration and white blood cell(WBC)count,and which in the ET patients was positively correlated with the platelet count.Conclusion FQ-PCR can quickly and accurately detect JAK2V617F mutation and its mutation ratio,which provides an effective laboratory indicator for the clinical diagnosis of CMPD.Certain relation exists between the JAK2V617F relative quantification and the clinical hematological indexes of the patients.

Objective To explore the effect of evidence-based nursing of the success rate of venipuncture for children and summarize the strategies . Method Three hundred and sixty children receiving intravenous therapy were investigated by self-designed questionnaire to explore the influencing factors of success veinpuncture. Results The unsuccessful rate of veinpuncture was 22.2%(88/360). And the main influencing factors included children′factors which accounted for 42.1%, nurses′factors which accounted for 29.4%, parents′factors which accounted for 18.1%and environment factors 10.4%. Conclusion The following strategies can be effective in increasing the success rate of veinpuncture, such as choosing the appropriate veins, improving the veinpuncture skills, creating favorable treatment environment and doing psychological nursing well.

Objective The expression of agr and sigB regulation system in Staphylococcus aureus with different infection types were assessed by analyzing the characteristics of m /z value generated by MALDI-TOF-MS.Methods A total of 50 isolates with specific genotypes were collected from Tianjin First Center Hospital during Jun 2013 to Feb 2014 for retrospective study .The pattern profiles of these isolates were obtained by MALDI-TOF-MS with RUO model, and the m/z value was also analysed to evaluate the expression the agr and sigB regulation system .The phylogenetic tree based on mass spectrum peak feature was constructed using SARAMIS software .Results A total of 50 strains of Staphylococcus aureus were divided into two groups: acute infection and chronic persistent and recurrent infection .The expression of delta toxin in acute infection and in chronic infection was 99.2 ±4.1 and 60.5 ±10.1 ( t =16.83, P<0.05), respectively.The regulation of stress proteins of sigB system was enhanced in chronic persistent and recurrent infections , and the expression intensities of SAS 030, SAS049 and SA0772 were 27.1 ±14.7, 54.8 ±21.5 and 51.6 ±19.2, respectively; while in acute infections , those were 4.9 ±1.9, 12.4 ±2.8 and 15.7 ±6.9, respectively.The t values between the two groups were -6.88 (P<0.05),-8.98 (P<0.05) and -1.87 (P<0.05), respecitively.The expression of phenol-soluble modulins (PSMs) was inconsistent , and the relative strength of PSMα3 was 100%in the colony variants small strains .Conclusions Different types of the Staphylococcus aureus infections could be evaluated through the assessment of the agr and sigB regulation system .The m/z value obtained by MALDI-TOF mass spectrometry is a marker for the expression of agr and sigB regulation system .The application of this technology needs further development .

Objective; To clone, sequence and analyze the coding sequences of the Mr 15000 and Mr 9000 active segments of natural granulysin derived from human CTLs activated by allogenic antigen ,in order to establish the basis for further purifying and investigating the immune - impairing mechanism of granulysin. Methods; The coding sequences of the Mr 15000 and Mr 9000 granulysin gene were amplified from the total RNA of activated CTLs of healthy person after reverse transcription by Nested - PCR, inserted into pET32a ( + ) vectors and then transformed into E. Coli TOP10, respectively. The recons were identified by PCR, endonuclease digestion and sequencing. Results: The whole coding sequences of the Mr 15000 and Mr 9000 active segments were successfully cloned as expected. The accurate pET32a - Mr 15000 and pET32a - Mr 9000 recons were obtained through Colony - PCR, endonuclease digestion and sequencing. There lied polymorphism on the 119 th amino acid of the product encoded by GLS gene. Conclusion; The coding sequences of the Mr 15000 and Mr 9000 active segments of human granulysin can be obtained and cloned by the methods mentioned above and can be used for subsequent research.

Objective To evaluate the method of PSM-mec detection by Vitek MS for nosocomialacquired methicillin-resistant Staphylococcus aureus (MRSA) identification.Methods Totally 167 isolates of MRSA and 100 isolates of methicillin-sensitive Staphylococcus aureus (MSSA) used in this research were non-repetitively and prospectively collected between June 2012 and December 2013,two different SCCmec genotyping methods were applied for the MRSA strains,Vitek MS was used for identification of the isolates,the acquisition mass-spectrogram and the result mass-spectrogram at Myla system were analyzed among the different SCCmec type of MRSA.Results The 167 isolates of MRSA were classified into 5 major SCCmec types,among which SCCmec Ⅰ accounting for 3.6％ (6 isolates);SCCmec Ⅱ 6.0％ (10 isolates);SCCmec Ⅲ and Ⅲa 84.4％ (141 isolates);SCCmec Ⅳand Ⅳ a 4.8％ (8 isolates);SCCmec Ⅴ 1.2％ (2 isolates),respectively.The peak adjacent to the horizontal axis of a m/z 2 500 could be visually identified between the SCCmec Ⅱ and Ⅲ MRSA,of which the delta toxin peak were presented at m/z 3 005-3 009 or m/z 3 037-3 056,while the strains without delta toxin peak and the other types of MRSA or MSSA had no characteristic peak at the same position.Conclusions Nosocomial-acquired MRSA of the drug-resistant condition could be rapidly differentiated and forecasted by Vitek MS.Vitek MS could serve as a routine clinical assistance for epidemiological investigations of nosocomial-acquired MRSA in local area.

BACKGROUND:Lysophosphatidic acid (LPA) had 3 kinds of receptors in vitro. Some researches had showed that LPA1, LPA2 and LPA3 receptors distributed widely in mouse cerebral cortex, nephritic external medulla layer and internal medulla layer, spermary, thymus, heart, lung, stomach, spleen, whereas less in liver, small intestine and skeletal muscles. Whether there are various LPA receptors in mouse VSMC membrane deserves further study.OBJECTIVE: To observe the mRNA expression of LPA receptors in vascular smooth muscle cells (VSMC).DESIGN: Observational comparative experiment.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: A total of 24 C57BL/6 mice aged 7-8 weeks, of either sex, with the body mass of approximately (25±3) g,were purchased from the Animal Department of Tongji Medical College, Huazhong University of Science and Technology. Trizol was purchased from America Invitrogen; dNTP Mix, Rnasin were obtained from TaKaRa; M-MLV reverse transcriptase and buffer system were from Promega; Taq DNA-polymerase and buffer system were from Biostar.Oligo(dT)18 primer were from Sangon, Shanghai. Primer sequences were designed referring to literature and nucleotide sequence database and synthesized by Sangon, Shanghai.METHODS: The experiment was conducted at the Comprehensive Laboratory, Tongji Medical College, Huazhong University of Science and Technology between August 2005 and January 2006. Mice were anaesthetized by abdominal cavity with 20 g/L ketamine (5 mL/kg). Thoracic aorta was obtained sterilely, and VSMC was cultured with adherence method. The 4th-6th passage cells were used in the trial. Cell purity was over 95%. Reverse transcriptase-polymerase chain reaction (RT-PCR) was applied to determine the mRNA expression of LPA receptors gene in mouse VSMC.MAIN OUTCOME MEASURES: Detection of mRNA expression of LPA receptors in mouse VSMC and comparison of receptor types.RESULTS: There were no significant differences between the expressions of LPA1 receptors and LPA2 receptors (P ＞ 0.05). Compared with LPA1 receptors and LPA2 receptors (0.79±0.05,0.82±0.06), the LPA3 receptor expression was lower (0.53±0.05, q =23.78,26.53, P＜ 0.01 ).CONCLUSION: There are 3 kinds of LPA receptors in VSMC, and their molecular masses are 600 bp, 463 bp and 899 bp,respectively. There are no differences for the expressions between LPA1 receptors and LPA2 receptors, while the LPA3 receptor expression is less.

Objective To observe the effect of proximal needling at Tianzhu (BL10) plus electroacupuncture at the Balance Zone on the lower-limb motor function and walking function in cerebral stroke patients.Method A hundred cerebral stroke patients were randomized into a rehabilitation group and a proximal needling group. The rehabilitation group was intervened by balance training and conventional rehabilitation exercises; the proximal needling group was intervened by proximal needling at Tianzhu plus electroacupuncture at the Balance Zone in addition to the treatment given to the rehabilitation group. The interventions weregiven once a day, 5 times a week, successively for 4 weeks. Prior to the intervention and after 4-week treatment, the lower-limb motor function and walking function were estimated by using lower-extremity Fugl-Meyer Assessment (FMA), time taken for 10-meter walk, and Holden's Functional Ambulation Classification (FAC).Result After 4-week treatment, the FMA, 10-meter walk time and FAC were significantly enhanced in both groups compared to the indexes prior to the treatment (P<0.01); the FMA, 10-meter walk time and FAC in the proximal needling group were significantly superior to those in the rehabilitation group (P<0.05).Conclusion In addition to conventional rehabilitation and acupuncture, proximal needling at Tianzhuplus electroacupuncture at the Balance Zone can increasethe lower-limb motor function score and promote the activities of daily life in cerebral stroke patients; it can also strengthen the security in walking and keep the patients from falling down.