BACKGROUND: Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a kind of cytokine which can stimulate the differentiation and proliferation of haematopoictic precursor cells and plays an important role in the process of immunoloregulation. Escherichia coli (E.coli) expression system has advantages,such as low cost and high output, over other systems.Therefore, E.coli is still the most commonly used exogenous gene expression system.OBJECTIVE: To observe the expression of hGM-CSF in the E.coli swain DH5α.DESIGN, TIME AND SETTING: The present observational experiment was performed at the Central Laboratory of Biotechnology Research,China Three Gorges University between February and June 2007. MATERIALS: Plasmid pBR322hGM-CSF was purchased from American Type Culture Collection, USA.hGM-CSF antibody was sourced from Sigma Company, USA.IPTG, X-gal,and vector pMGT-18 were purchased from Shanghai Bioengineering Technology Company, China. METHODS: Primer was designed according to hGM-CSF gene. Taking cloning vector plasmid pBR322-hGM-CSF as template, hGM-CSF gene was acquired and recombined into prokaryotic expression vector pGEX-4T-1. Recombinant plasmid confirmed by enzyme digestion and sequencing was transformed into E.coli strain DH5α and induced by isopropy-β-D-thiogalactoside (IPTG). Expression product was identified by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and detected by Western blotting assay. MAIN OUTCOME MEASURES: hGM-CSF expression in the E.coli strain DH5α. RESULTS: SDS-PAGE results revealed that the recombinant hGM-CSF protein with relative molecular weight of 40 500 was produced up to approximately 17.9% of total protein. Western blotting detection results indicated that there was a 40 500 bp brand. CONCLUSION: The present study reconstructed prokaryotic expression vector pGEX-hGM-CSF, induced its expression in the E.coli strain DH5α, and obtained hGM-CSF fusion protein.

Objective To select the aptamer to an extracellular soluble fragment of recombinant human TGF-? receptor Ⅱ (TGF-? RⅡ) in order to antagonize TGF-? effectively by using systematic evolution of ligants by exponential enrichment (SELEX). Methods Initial random RNA library was transcripted in vitro from ssDNA 5′-TAATACGACTCACTATAGGGAGGACGATGCGG-N60-CAGACGACTCCCCGA-3′; rhTGF-? RⅡ was used as target protein. Totally,selection of 8 times was carried out in SELEX experiment. Membrane binding assay was performed to detect the evolution of enriched RNA library; Electrophoretic mobility shift assay (EMSA) was done to determine the affinity between the selected nucleic acid sequence and TGF-? RⅡ. Results Evolution of the enriched RNA library along the increased affinity to TGF-? RⅡ was observed with the development of selection. Two types of dominant sequences were isolated and named as sequence A and sequence B. In membrane binding assay,both sequences A and B showed obvious affinity to TGF-? RⅡ. However,no retarded bands were seen in EMSA. Conclusion The affinity of sequences A and B to TGF-? RⅡ is beyond satisfaction. However,possible sequences with improved affinity to TGF-? RⅡ can be selected by post-SELEX on the basis of candidate sequences A and B.

This study was aimed to investigate the preventing efficacy of the combined using of Er-Shi-Wu-Wei Shan-Hu (ESWWSH) pill, Ru-Yi Zhen-Bao (RYZB) pill, Er-Shi-Wei Chen-Xiang (ESWCX) pill on the focal cere-bral ischemia injury in rats. A total of 80 Sprague-Dawley (SD) rats were randomly divided into four groups. In the sham operation group and the MCAO model group, normal saline was given orally every day (1 mL/100 g). In the positive control group, nimodipine was given orally every day (1.89 mg/100 g). In the drug combination group, ESWWSH pill (7 mg/100 g), RYZB pill (31.5 mg/100 g), ESWCX pill (49 mg/100 g), were separately given in the morning, noon and evening every day. Improved MCAO was used in the model establishment after 7-day intragastri-cal administration. After 24 h brain infarction, TTC method was used in the determination of the cerebral infarct size. Drying and weighing method was used in the calculation of brain water content. Determinations were made on con-tent of NO in ischemic brain issues (nitrate reductase method); activities of SOD in serum (xanthine oxidase method);content of MDA in serum (TBA method); and activities of LDH in serum (pyruvate method). The results showed that compared with the MCAO model group, the combined using of three Tibetan classical prescriptions can obviously re-duce the cerebral infarct size, brain water content and the NO content in brain issues, increase the activity of SOD, reduce the content of MDA, and reduce the activity of LDH in serum. It was concluded that the research preliminari-ly showed that the combined using of ESWWSH pill, RYZB pill and ESWCX pill can protect the ischemic brain tis-sues, such as reducing the cerebral infarct size, alleviating brain edema, and inhibiting oxidative stress reactions.

Objective:To study the clinicopathologic characteristic of the primary extranodular non-Hodgkin's lymphoma in head and neck.Method:Clinical manifestation and the characteristic of clinicopathology of 216 extranodular non-Hodgkin's lymphoma patients in head and neck were analyzed retrospectively.Result:The age of thirty-one to sixty and seventy-one to eighty years old were more common age range in the group of patients. Nasal cavity was the most commonly primary involved site(95 cases, 44.0% ) , secondly was tonsil ( 47 cases, 21.8%). The most common histologic subtype was NK/T cell lymphoma, which accounted for 49.5%(107 cases)of cases, secondly was diffuse large B cell lymphoma (DLBCL, 58 cases, 26.7%). The most common histologic subtype in different swtach groups respectively is: NK/T cell lymphoma in nasal cavity(74 cases,77.9%), DLBCL in paranasal sinus(6 cases,50.0%), DLBCL in tonsil(27 cases,57.4%),NK/T cell lymphoma in nasopharynx(17 cases,44.7%), and DLBCL in lingual root(5 cases,45.4%).Conclusion:We conclude that primary extranodular non-Hodgkin's lymphoma is common in head and neck patients. There is characteristic in age, primary involved site and histologic subtype, which is helpful to understand these characteristic for pathologic diagnosis.

Objective To further explore the relationship between increasing genetic material and clinical manifestation of partial trisomy 5p .Methods G‐banding karyotypes of peripheral blood lymphocytes in the patient and his parents ,and at the same time to summary the partial trisomy 5p clinical performance .Results patient ,46 ,XX ,der(6)t(5 ;6)(p13;q25) mat ;partial trisomy for 5p13→pter resulting from the balanced translocation of the mother .Mother:46 ,XX ,t(5;6)(p13 ;q25);carrier of a balanced 5/6 translocation .Father :46 ,XY .Conclusion The phenotype of trisomy 5p may be associated with express and function of gene at spe‐cial chromosome region .

Objective To discuss the application value of DWI and ADC on predicting therapeutic effect of radiotherapy treatment in NPC. Methods Twenty four local recurrent cases and 38 non-recurrent cases after radiotherapy treatment in NPC were reviewed. MRI and DWI-MRI were performed at pre-radiotherapy and 3, 6, 12 months after treatment, the ADC values of the lesions were analyzed by SPSS 18.0 statistical software. ROC curves based on the ADC values were measured in 3, 6, 12 months after treatment plotted to analyze the threshold ADC value for confirming recurrence. Results The recurrent group and newly diagnosed group showed significantly high signal on DWI, while the non-recurrent group acquired low or mixed signal. The ADC values of the primary tumor in the recurrent group and the non-recurrent group were (0.709 ± 0.078) × 10-3 and (0.693 ± 0.089) × 10-3mm2/s, respectively, t=-0.717,P>0.05, respectively.The ADC values of the primary and recurrent tumor in the recurrent group were (0.730± 0.068) × 10-3mm2/s and (0.709 ± 0.078) × 10-3mm2/s, t=-1.000,P>0.05 , respectively.There were statistical differences between the recurrent group and the non-recurrent group for ADC in 3, 6, 12 months after treatment:(1.128 ± 0.179) × 10-3 and (1.358 ± 0.145) × 10-3mm2/s, t=5.567,P<0.01;(1.164 ± 0.174) and (1.450 ± 0.102) × 10-3mm2/s, t=7.310,P<0.01;(1.107 ± 0.180) × 10-3 and (1.584 ± 0.125) × 10-3mm2/s, t=11.189,P<0.01;respectively. Take 1.29 × 10-3 mm2/s,1.32 × 10-3mm2/s,1.37 × 10-3mm2/s respectively in 3, 6, 12months after treatment as the diagnostic threshold to predict tumor recurrence. The sensitive , specificity, and accuracy were (83.3%, 73.7%, 77.4%), (83.3%, 89.5%, 87.1%), (100.0%, 94.7%, 96.3%).Conclusions Both DWI and ADC value are important for diagnosing and predicting recurrent NPC after radiotherapy treatment, DWI and ADC can be used to regular follow-up after radiotherapy, to further improve the rate of early diagnosis of recurrent NPC.

Objective To investigate the effect of the artificial cycle on the prognosis after transcervical resection of adhesion (TCRA) and its mechanism .Methods 80 patients with intrauterine adhesion were randomly divided into the observation group and the control group .The observation group were performed TRCA and postoperative artificial cycle for successive 3 months :oral es-tradiol valerate(9 mg once daily) for 21 d ,adding oral medroxyprofgesterone acetate(10 mg once dialy) on 15 d ,then the medication discontinuation for 7 d ,which was taken as 1 cycle;the control group received only TCRA .The re-examination was performed after 3 months .The levels of follicle stimulating hormone(FSH) ,luteinizing hormone(LH) ,estradiol(E2 ) ,prolactin(PRL) ,progesterone (P) and testosterone(T) were examined by ELISA after TCRA three months .The expression of MMP-9 and TGF-β1 in adhesive endometrium were detected by real-time PCR and Western blot .Results Compared with the control group ,the artificial cycle thera-py combined with the operation could significantly increase the effective rate of TCRA for treating mild and moderate intrauterine adhesion(P<0 .05);the seum levels of FSH ,LH ,E2 ,PRL ,P and T had no statistical differences between the observation group and the control group(P>0 .05);the artificial cycle significantly improved the adhesive degree of intrauterine adhesion ,increased the MMP-9 expression and decreased the TGF-β1 expression .Conclusion Artificial cycle could improve the degree of uterine cavity ad-hesion after TCRA ,which the mechanism might be related to the increase of MMP-9 expression and decrease of TGF-β1 expres-sion .

Objective To explore the effect of early stage of comprehensive lung rehabilitation on cervical spinal cord injury in patients with mechanical ventilation.Methods The patients with cervical spinal cord injury admitted from January to December 2015 were selected as the control group which was treated with routine nursing measures.The patients with cervical spinal cord injury admitted from January to December 2016 were selected as the treatment group,which received early stage lung rehabilitation measures.We compared two groups of patients in the using time of breathing machine,the total length of hospital stay and pulmonary infection rate.Results The duration of mechanical ventilation and hospitalization days in the control group were(13.98±11.11),(25.10±16.98)d,which were higher than those in treatment group[(7.23±4.94),(16.26±6.41)d](P<0.05).The infection rates of control group and treatment group were 76.19％and 52.38％respectively,and the difference was significant(P<0.05).Conclusion In the early stages comprehensive pulmonary rehabilitation measures for mechanical ventilation in patients with cervical spinal injury can shorten the breathing machine use time and the total hospitalization days,and reduce pulmonary infection rate.

OBJECTIVE: To probe into the standardized management of dispensing in the pharmacy METHODS: To introduce the idea of standardization in dispensing, to establish and implement SOP(standard operating procedure), to carry out networking management, to setup the modern facility, and to introduce the new open and divisional mode for dispensing drugs and to put dispensing and supply of drugs in over - inclusive standardized management .RESULTS: The standardized management of dispensing and supply of drugs was achieved on the whole and the aim of precise dispensing and scientific management was fulfilled. CONCLUSION: The establishment and implementation of SOP, the networking management of drugs and mord-ernization of facility are the basis of standardization of dispensing of drugs.