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 .

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 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.

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.

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 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.

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 study the appearance of MR diffusion weighted imaging in early stages of cartilage degeneration and to detect its values. Methods In 20 goat left knees, intra- articular injection of 5 units of papain was performed causing a loss of cartilage proteoglycan. Twenty right knees were used as control group. MR diffusion weighted imaging was performed at 24 hours after intra-articular injection of papain. ADC of each part of articular cartilage was measured and compared with each other. The proteoglycan content was measured biochemically and histochemicaUy. Routine MRI and DWI were performed in 100 patients with osteoarthritis and 20 healthy people. The ADC of each interested part of articular cartilage was measured and compared with each other. Results In experimental control group, the ADCav of articular cartilage was (14.2±2.3)×10-4 mm2/s. In early stages of cartilage degeneration group, the ADCav of articular cartilage was (17.5±4.2) × 10-4 mm2/s. The ADCav of the control group was lower than that of the early stages of cartilage degeneration group (t = 2.709 ; P = 0.016) . The proteloglyean content of articular cartilage was 4.22×10<'6> μg/kg in control group, and 0.82×10<'6>μg/kg in experimental group at 24 hours after injection of papain. The difference between control group and experimental group was significant (t = 2.705, P = 0.018). In healthy people, the ADCav of articular cartilage was (7.6±2. 2) × 10-4 mm2/s. In osteoarthritis group, the ADCav of articular cartilage was (10.3±4. 2) × 10-4 mm2/s. The ADCav in the healthy group was significantly lower than that in the osteoarthritis group (t = 2.609, P = 0.014). Conclusion DWI is an useful method in detecting early stages of cartilage degeneration which can not be showed on routine sequences.

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.