Objective To construct GST/HIF1α fusion protein expression vector and induce its expression in Escherichia coli (E.coli). Methods The coding sequence of hypoxia inducible factor-la (HIF-la) and its deletion fragments were amplified from the plasmid pcD-NA3.1-HIF-la by PCR and inserted into pGEX-4T-2 by BamHI and Not I. The positive recombinanls were identified by restriction endonu-clease digestion and DNA sequencing. Then they were transformed into E.coli BL21, induced by IPTG and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmid pGEX-4T-2-HIF-lα and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing. The desired CST/HIF-1α fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid of HIF-la and its deletion mutants were successfully constructed and the expression of fu-sion proteins was confirmed. This study provides the basis for the further research on purifying HIF-la protein and the biological function of HIF-lα.

Objective: To establish the optimum preparation process for Suxiaoxintong Dropping Pills (Cortex moutan, Rhizoma Chuanxiong, Borneolum Syntheticum). Methods: The preparation conditions, such as the different kinds of primary substance, cooling agent, the proportion between the extracts and primary substance etc. were studied with L 18 (3 7) orthogonal design. Results: Six batches of the products prepared with the process on the experimental result were conformed to the standard of the Chinese pharmacopoeia. Conclusion: It has been proved that the process is available.

ObjectiveTo discuss the relationship between sit-to-stand velocity and dynamic and static balance respectively in stroke patients.Methods22 stroke patients were measured their time for Five Times Sit to Stand Test (FTSTS scores) and the static balance performance as well as swaying of center of gravity in lateral directions was recorded with GS-10A. In addition, the distances of functional reach test (FRT) when sitting were measured.ResultsBy using Pearson's correlation coefficient, FTSTS scores showed the moderate correlation with hemiparetic side, ENV. Area and distance of sitting FRT (r=-0.512, P<0.05；r=0.632, P<0.05；r=-0.662, P<0.01). Linear regression model, in terms of balance ability, showed the main factors associated with the sit to stand velocity were the distance of sitting FRT and ENV.Area. ConclusionENV.Area and distance of sitting FRT are the main factors associated with sit to stand velocity and both static and dynamic balance can predict sit to stand velocity.

Objective To analyze the test results of persons with noise -induced hearing loss using disyllabic Mandarin speech test materials (MSTMs) .Methods 57 subjects with noise -induced hearing loss (noise group) were divided into three groups according to the duration of noise exposure :less than 10 years ,between 11 and 20 years and more than 20 years ,and also separated into 2 groups in terms of the degrees of hearing loss (group A :pure tone average thresholds of 0 .5 ,1 ,2 ,and 4 kHz less than or equal to 30 dB HL ,group B:pure tone average threshold of 0 .5 ,1 ,2 ,and 4 kHz greater than 30 dB HL) .66 subjects with normal hearing were in the control group .All the subjects enrolled in this study could speak Mandarin well in their daily life .Nine lists of disyllabic Mandarin speech test materials were utilized to test SRT and P -I function for these groups respectively .Results PTA and SRT in the noise group were much higher than the control group ,the average slope of P -I function was 5 .94% /dB ,and the curve of P-I function moved to the right side .SRT showed obvious changes as the increase of the years of noise exposure and especially in the group which had more than 20 years noise exposure (P<0 .01) . SRT of group B which had more hearing loss were greater than that of group A (P<0 .01) .Conclusion SRTs in the noise group are much higher than the control group with the average slope of P -I functions decreasing and the curve moving to the right side .As the years of noise exposure increase and the degrees of hearing loss become more severe ,SRT rise more apparently .

Objective To clone and express the gene encoding Schistosoma japonicum myophilin-like protein (SjcMLP) and to study the antigenicity of the recombinant protein. Methods The SjcMLP gene was amplified by PCR. The PCR product was cloned into T vector, and then subcloned into expression vector pQE30. The recombinant plasmid of pQE30-SjcMLP was transformed into E.coli M15, and induced with IPTG for expression. The bacterial lysis was conducted by ultrasonication and the supernatant was analysed by SDS-PAGE. The recombinant protein (reSjcMLP)was purified with the Ni-NTA resin, and analysed with SDS-PAGE and Western blot. The titers of sera from C57BL/6 mice immunized subcutaneously with reSjcMLP were detected by ELISA. Results The results of SDS-PAGE and Western blot showed that the molecular weight of expressed fusion protein was around 24.8 kDa and was recognized by the sera from the mice infected with Schistosoma japonicum. The purified protein of reSjcMLP was coated for ELISA test and the IgG titers in the sera from the mice immunized with reSjcMLP were as high as 1∶12 800 reacted with. However, no significant difference was found in worm reduction rates between the immunized mice and control mice. Conclusions The fused recombinant protein of reSjcMLP is successfully ex-pressed and purified. The recombinant protein in this experiment fails to induce significant protection against the challenge infection in C57BL/6 mice.

Invasive neural electrodes promote human understanding of neuroscience to the micrometer and millisecond scale. Due to the large mechanical mismatch between traditional rigid electrodes and soft brain tissues, flexible electrodes have become the new trend of neural electrodes. The flexibility of the neural electrode reduces the immune response while losing the implantation stiffness. The implantation mechanism of the neural electrode was reviewed and current researches on the implantation strategies of the flexible electrodes were summarized, so as to help solve the loss of implantation ability of flexible electrode and acute implantation injury. Based on the characteristics of various implantation strategies, the prospect of flexible electrode implantation strategies was proposed.

pUDK-HGF, the recombinant plasmid DNA encoding human hepatocyte growth factor (HGF), can treat ischaemic disease. A great quantity of pharmaceutical pUDK-HGF is needed. A pilot-scale production process of pUDK-HGF was established based on a new chromatographic media (plasmidselect), including fermentation, cell harvesting, alkaline lysis, ultrafiltration, RNA removing and buffer exchanging on Sephacryl S-1000, capturing supercoiled plasmid DNA with plasmidselect, and removing the salt with Sepharose 6BFF. The process does not use RNase enzyme and toxic solvents.
DNA, Recombinant ; biosynthesis ; DNA, Superhelical ; isolation & purification ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Genetic Vectors ; genetics ; Hepatocyte Growth Factor ; biosynthesis ; genetics ; Humans ; Pilot Projects ; Plasmids ; isolation & purification

Objective To explore safety evaluation of the approaches of the percutaneous eilational tracheostomy(PDT)ane traeitional tracheotomy in the treatment of neurological patients. Methods The stuey eesign was a multicenter,prospective,raneomizee clinical trial. One huneree ane seventy-six cases with acute nerve trachea incision from Feb. 2010 to Feb. 2013 of 3 hospitals were selectee as our subject. They were raneomly eivieee into the traeitional group ane PDT group. The information inclueing operation time,the incieence of pneumothorax,subcutaneous emphysema,tracheal fistula,esophageal,trachea ane lung injury from complications such as infection were recoreee. Results The complication rate in traeitional group was 19. 51%(16 / 82),higher than that of PDT group(8. 51%(8 / 94),P = 0. 021). The surgery perioe in PDT group was(7. 5 ± 2. 3)min,shortee than that in traeitional group((41. 6 ± 5. 8)min,P = 0. 000). Conclusion The approach of percutaneous tracheostomy can quickly buile airway of neurological patients with character of simple, safe,ane it also can reeuce the incieence of respiratory complications.

Metabolic associated fatty liver disease （MAFLD） has become the most common liver disease in the world， including simple hepatic steatosis， steatohepatitis， liver fibrosis/cirrhosis， and liver cancer， and it is often associated with metabolic syndrome. MAFLD has complex pathological manifestations， and there are still no new drugs on the market in China. In the past， traditional Chinese medicine often treated this disease from the perspective of food， phlegm-dampness， damp-heat， or liver depression and spleen deficiency， but a detailed analysis showed that the pathological process of the disease was consistent with the theory of “six stagnation diseases” in traditional Chinese medicine. This article briefly introduces the origin of the pathogenic theory of “six stagnation diseases”， the pathological features of the “six stagnation diseases” leading to fatty liver disease， the key points of syndrome differentiation and treatment， and clinical practice， so as to promote clinical treatment through an in-depth understanding of the traditional Chinese medicine pathogenesis of MAFLD.