Objective To explore the expression changes of HSP90αin cardiac muscles and survival rates in rats by using the different fluids to resuscitate the severs hemorrhagic shocked rats ,and provide reference for the clinical treatment of hemorrhagic shock with different resuscitation fluids .Methods Uncontrolled hemorrhagic shock rats model was established ,using lactic acid salinger liquid ,poly peptide injection gelatin ,hypertonic sodium chloride dextran for fluid resuscitation respectively ,and then checked the HSP90αexpression changes and survival rates in rats .Results the expressions of HSP90αin myocardial tissue and the mortality in rats were different after using different resuscitation fluids in severe hemorrhagic shock rats ,difference was statistically significant(P<0 .05) .Conclusion the expression of HSP90α in cardiac muscles of rats could be induced by severe hemorrhagic shock ,the HSP90αexpressed differently and regularly after using different resuscitating fluids ,it implied that the HSP90α played an important role in the hemorrhagic rats cardiac as a regulating fator .

AIM: To investigate the effect of basic fibroblast growth factor (bFGF) on radiation-induced splenocytic apoptosis of mice. METHODS: At 14 h after whole body irradiation with 0.5 Gy and 1.0 Gy,splenocytes were cultured with and without bFGF,and splenocytic apoptosis was quantitatively analysed by flow cytometry. Cell proliferation was determined by the method of [3H]-TdR incorporation. RESULTS: bFGF(1 ?g/mL and 2 ?g/mL) could reduce the rate of cell apoptosis ,and promote the proliferation of splenocytes. CONCLUSION: bFGF could inhibit radiation-induced splenocytic apoptosis and promote the proliferation of splenocytes and then enhance body immunity.

Objective To explore the relationship between condition of hypoxia and prognosis in patient with renal clear cell carcinoma (RCCC).Methods The expression of hypoxia-inducible factor-lo( HIF-1 α) protein in cancer tissue of 89 RCCC cases was examined by streptavidin-biotin complex immunohistochemistry.Clinical and pathological data and prognosis of 89 cases were analyzed retrospectively.There were 66 males and 23 females,with an average age of 57 years.The patients were divided into two groups,the chronic pulmonary disease (CPD) group ( 19 cases) and non CPD (NCPD) group (70 cases),with 46cases in clinical stage I,15 cases in stage Ⅱ,26 cases in stage Ⅲ,and 2 cases in stage Ⅳ.The relationship between survival time and clinicopathological variables including the presence of CPD,the positive rate of HIF-1α protein,smoking history and hemoglobin level were evaluated by the Kaplan-Meier method.And the Cox proportional hazards regression model was build to analyze the correlation between each variable and survival time.Results The 89 cases were followed up for a median time of 19 months (6 to 84 months).Twenty cases died,and 69 cases survived.Between the CPD group and NCPD group,clinical stage,hemoglobin level and the degree of expression of HIF-1 α were significantly different (P ＜ 0.05 ).The median survival time was 44 and 71 months in CPD group and NCPD group,respectively,and the difference was significant ( P ＜ 0.05 ).The median survival time was 43 and 70 months in Hb≤ 110 g/L group and Hb ＞ 110 g/L group,respectively,and the difference was significant ( P ＜ 0.05).The stronger the degree of expression of HIF-lα was,the shorter the overall survival was.And the difference was significant ( P ＜0.05 ).Associated with CPD,hemoglobin level,the expression of H1F-1α were independent factors affecting overall survival of the patients (P ＜0.05 ).CPD and HIF-1 α expression were positively related to disease-free survival time,and hemoglobin level was negatively related to disease-free survival time.Conclusions Systemic hypoxia caused by CPD may aggravate the hypoxie state in the organization of the patients with RCCC.The condition of hypoxia and prognosis in patient with RCCC is negatively correlated.

In this paper, the biological compatibilities of FGF/Collagen compound sponges are evaluated. Intracutaneous stimulation test and shortterm muscle embedding test of FGF/Collagen compound sponges made of different materials are performed. Experimental data are analyzed and evaluated according to the standard. The two sponges get marks of 0 in acute eye stimulation test and intracutaneous stimulation test, thus they are not stimulating to tissues. In shortterm muscle embedding test, they don't lead to inflammation reactions and can be degraded and absorbed in short term. FGF/Collagen compound sponge proves good biological compatibility and has a cheerful prospect in medicine.

Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

AIM: To investigate the pharmacological effects of recombined basic fibroblast growth factor (rbFGF) on retinal ganglion cells (RGCs) of rats. METHODS: Using calibrated cross-action forceps a moderate crush injury was inflicted on the nerve. After crush injury, rbFGF, saline and VB 12 were administered by retrobublar injection. Four weeks after injury , the apoptosis of RGCs was measured with flow cytometer. RESULTS: Four weeks after operation, it was shown that the rbFGF, but not saline or VB 12 injection could significantly improve the maintainance of RGCs of rats. After 800 U, 1600 U and 2400 U rbFGF injection, the injured RGCs were rescued by 24.5%, 27.3% and 28.5% respectively. Furthermore, it was also found that rbFGF injection could effectively prevent the axons from injury. The flow cytometer showed that the rate of apoptosis was reduced markedly on 7 days at rbFGF group. CONCLUSION: rbFGF can significantly promote the functional repair of injured optic nerve. [

Objective:To establish the method for detecting lower respiratory infections (LRIs) bacterialpathogens using nanopore sequencing, and evaluate the feasibility of this method.Methods:Bronchoalveolar lavage fluid (BALF) samples from 33 patients with LRIs who visited the Department of Respiratory and Critical Care Medicine of Beijing Hospital from July 2019 to September 2020 were collected.Nanopore 16S amplicon sequencing were performed on these samples. In order to evaluate the clinical value of the nanopore sequencing, χ 2 test was used to analyze the pathogen differences between the detection rate and pathogen types results found with using the nanopore 16S sequencing and the results found with bacterial culture. Results:The process and method of nanopore sequencing used in the detection of the LRIs pathogens were established. The pathogen detection rate of the 16S sequencing was higher than that of the traditional bacterial culture (75.8% [25/33], 45.5% [15/33], χ2=5.140, P<0.05). From the 25 positive samples found with nanopore 16S sequencing, 16 pathogens were detected, including Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, Streptomonas maltophilia, Acinetobacter baumannii, and Acinetobacter junii, Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecalis, Enterococcus gallinarum, Corynebacterium striatum, Mycobacterium paraintracellulare, Serratia marcescens, Achromobacter insuavis, Citrobacter murliniae and Mycoplasma pneumoniae. More than 6 pathogens were tested in clinical culture, including Haemophilus parainfluenzae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Streptomonas maltophilia (χ2=7.949, P<0.05). 16S sequencing aligned to species level sequences accounted for 80.0 (60.0, 86.0)% of the genus level. The results obtained by using16S sequencing and bacterial culture were consistent in 11 (33.3%) samples. Conclusions:Nanopore 16S amplicon sequencing can quickly identify pathogenic bacteria from BALF in LRIs patients. Nanopore 16S amplicon sequencing has a high detection rate, it can detect more pathogens than traditional bacterial culture, and it can also identify most bacteria to the species level. This technology is a very promising platform with broad application prospects.

Objective To build the suitable index system of the clinical research program acceptance evaluation.Methods The Delphi method and the weighted average method were adopted to determine the weight of each index.Results Followed by two rounds of Delphi method, twenty five evaluation indices were selected, including three first-class indices, seven second-class indices and fifteen third-class indices, and the weight of each index was calculated.Conclusions The index system,simple and reasonable, reflects the characteristics of clinical applied researches to a certain extent with a certain degree of operability, laid the foundation for the further supporting to carry out scientific research projects.

Objective:To express mycobacterium tuberculosis protein MPB 64 in baculovirus insect cell expression system ,and identify its immunogenicity.Methods:The target gene MPB64 was connected to pFastBac vector ,then the pFastBac-MPB64 plasmid which was harvested would transformed to DH10Bac competent,and the target gene was transposition into Bacmid by Tn7 transposase fragment,therefore Bacmid-MPB64 Shuttle vector was obtained.The shuttle vector was packaged by liposomes and transfected Sf 9 cells to harvest P1-generation virus ,then high titers of P4 generation virus was harvested by repeat transfected Sf 9 cells three times.The target protein MPB64 was purified from the supernatant by Q Sepharose FF and Ni affine chromatography ,which were used to immunize BALB/c mice.Antibody changes in serum would be detected ,and the proliferation of immunized mice spleen cells would be detected by MTT,detected the IFN-γsecretion by MPB64 stimulated spleen cells by ELISA method.Results: MPB64 successfully expressed in insect cells.The purity of target protein was over 90% and yield up to 35 mg/L after purification.Purified protein can effectively stimulate BALB/c mice to produce antibodies , increase the content of IFN-γmedium in mice spleen cells ,and significantly promoting proliferation in spleen cells between 0.2-100 μg/ml.Conclusion: MPB64 which has immunogenicity was successfully expressed in baculovirus insect cell expression system ,that open a new avenue for tuberculosis vaccine production.

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.