Objective To explore the regulation role and expression level of thymosin for Th1/Th2 and T-bet , GATA-3 in asthma model rats respectively.Methods 60 SPF grade male rats (6-8w) were randomly divided into six groups:control group, asthma model group, dexamethasone (0.5 mg/kg) group, thymosin high dose group (5 mg/kg), thymosin middle dose group(2.5 mg/kg), thymosin low dose group (1.25 mg/kg).The asthma model was constructed with ovalbumin.The rat in the control group and asthma model group were injected with normal saline by intraperitoneal , while other rats were injected with corresponding drugs.Then the concentration of IL-4 and IFN-γin the serum of rats were tested by the method of enzyme linked immunosorbent assay(ELISA) and the protein expression level of GATA-3 and T-bet factor in the lung tissue were tested by immunohistochemistry in each group.Results The concentration of IL-4 in the serum was higher and IFN-γwas lower in the asthma model group than the other groups , while the protein expression level of GATA-3 was higher and T-bet was lower in the asthma model group than the other groups ( all P <0.05 ) . Conclusion Thymosin upregulation of IFN-γand inhibition of IL-4 may be achieved by upregulating T-bet and downregulating GATA-3, and thymosin has therapeutic effect on asthma rats.

BACKGROUND: Now, the current study has not yet confirmed two treatment methods of the proximal femoral nail antirotation and total hip arthroplasty for treating intertrochanteric fractures in the elderly, which method has more advantages, and there is no clear conclusion at present. OBJECTIVE: To systematical y compare the repair effect of proximal femoral nail antirotation and total hip arthroplasty for treatment of intertrochanteric fractures in the elderly. METHODS: We searched the related literatures from 2011 to 2015 on Wanfang database, PubMed, MEDLINE and Embase database by computer. We retrieved the journals in China by hand and col ected randomized control ed trials on proximal femoral nail antirotation and total hip arthroplasty in the treatment of intertrochanteric fractures in the elderly. Intraoperative blood loss, operation time, hospitalization time, ambulation time after operation, and Harris score were selected as evaluation indexes. Data were analyzed using RevMan 5.2 software. RESULTS AND CONCLUSION: Final y, 7 Chinese literatures were included, with 811 patients. The results of meta analysis showed that the hospitalization time and ambulation time after operation were significantly shorter in the total hip arthroplasty group than in the proximal femoral nail antirotation group (P < 0.000 01), which reduced the pain of patients. Operation time and intraoperative blood loss were poorer in the total hip arthroplasty group than in the proximal femoral nail antirotation group (P < 0.000 01). No significant difference in Harris hip score was detected between the two groups (P > 0.05). These results confirmed that the internal fixation of proximal femoral nail antirotation in minimal y invasive therapy when reducing operation time and blood loss has a certain advantage. The total hip arthroplasty can make the recovery of hip function earlier, al ow early weight-bearing walking, reduce the patients’ bedridden time and reduce the complications in bed. Thus, the clinicians in the practical work should choose the appropriate therapy with considering the patient’s condition, damage degree, fracture type, and financial capability.

Aged ; Aortic Valve Insufficiency ; diagnostic imaging ; Echocardiography ; Humans ; Male

Objective To study the predictive value of serum hyaluronic acid (HA) levels for morphological progression of osteoarthritis (OA).Methods Seventy New Zealand white rabbits were randomly and equally divided into the OA group and the normal control group.OA was induced by injecting 0.5 ml of 2％ papain into the left knee joints and the same volume of sterile saline solution was injected Into right knee joints in control group.The serum HA levels were detected at 1,2,4,6,8,10 and 12 weeks after injection.Then magnetic resonance imaging (MRI) and pathohistological changes were observed.T test was used for the analysis of measurment.Resuts Compared with the control group,degeneration changes were found in the OA group,such as thinner articular cartilage,fibrillation and destroyed cartilage matrix,and inflammation,proliferation and degeneration of the synovial tissue.All these changes were much worse with prolonged observation time.The serum HA levels were increased [12 week:(657±132) ng/ml,vs (166±8) ng/ml],which correlate with structural damage.Conclusion Serum HA levels had a predictive value for further development of OA.

Educational Measurement ; Humans ; Inservice Training ; Pneumoconiosis ; diagnosis ; Reference Standards

Corynebacterium glutamicum SA001 is a mutant with lactate dehydrogenase (ldhA) deletion. In order to increase metabolic flux from isocitrate to succinate, and to improve the production of succinate under anaerobic conditions,we transducted the gene aceA coding isocitrate lyase (ICL) from Escherichia coli K12 into Corynebacterium glutamicum SA001 (SA001/pXMJ19-aceA). After 12 h aerobic induction by adding 0.8 mmol/L of IPTG, the recombinant strain was transferred to anaerobic fermentation for 16 h. Succinate reached 14.84 g/L, with a productivity of 0.83 g/(L x h). Compared to C. glutamicum SA001, the activity of ICL of the recombinant strain was increased 5.8-fold, and the succinate productivity was increased 48%. Overexpression of isocitrate lyase will increase the metabolic flux of glyoxylate bypass flowing to succinate.
Corynebacterium glutamicum ; genetics ; metabolism ; Escherichia coli ; enzymology ; genetics ; Gene Deletion ; Industrial Microbiology ; Isocitrate Lyase ; biosynthesis ; genetics ; L-Lactate Dehydrogenase ; genetics ; Succinic Acid ; metabolism ; Transduction, Genetic

Objective To develop an efficient recombinant expression system of alanine aminotransferase ( ALT ) in order to build the foundation for the preparation of feedstock related ALT reference materials.Methods A new human ALT gene was synthesized by optimizing the codons of the nucleic acid sequence encoding human ALT using bioinformatic tools, and then it was cloned into pRSF-Duet expression vector.The recombinant plasmid pRSF-Duet-ALT was transformed into E.coli BL21 and the target protein expression was induced by 2 mmol/L isopropyl β-D-1-thiogalactopyranoside ( IPTG ) .The expression condition for soluble protein was optimized by changing the inducer concentration, shaking speed and induction temperature. The soluble protein was purified by nickel ion affinity chromatography and dextran molecular sieve chromatography, and identified by sodium dodecyl sulfate-polyacrylamide gelelectrophoresis( SDS-PAGE) and Western blot analysis.The activity and stability of recombinant proteins in serum matrix under different storage conditions were detected.Results The usage frequency in E.coli of ALT codons was more than 10%after codon optimization.The expressions of soluble proteins were increased by optimizing the induced expression conditions, including a final concentration of 2 mmol/L of IPTG, and continued incubation with shaking at 150 rpm for 8 h at 25℃.The purified protein was identified as ALT by SDS-PAGE and Western blot with ALT activities of up to 80 000 U/L.Recombinant ALT could be stable for 2-8 d at 2-8 ℃or 25 ℃with a relative standard deviation of less than 5%.Conclusions An efficient recombinant expression system of ALT was developed successfully by codons optimization.The obtained recombinant protein could achieve the requirements of reference material feedstock.

Objective To investigate the role of p38MAPK in the differentiation of murine osteoblasts, and to observe the expressions of receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG). Methods The calvarial osteoblasts of newborn BALB/c mice were cultured in MEM medium containing 10% NCS. Raloxifene (10~(-7) mol/L) and 17β-estradiol (10~(-8) mol/L) were added respectively when cells reached 70%-80% confluence combined with or without 5 μmol/L SB202190, an inhibitor of p38MAPK. The osteoblasts alkaline phosphatase activity assays were performed 72 hours later using PNPP method, and mRNA levels of alkaliphosphatase (ALP), OPG and RANKL were determined by RT-PCR. Results 17β-estradiol and raloxifene increased ALP activity and ALP mRNA level in osteoblasts in vitro which were blocked by p38MAPK inhibitor.The mRNA levels of RANKL and OPG were up-regulated by 17β-estradiol and raloxifene while the ratio of OPG/RANKL kept constant. SB202190 (5 μmol/L) inhibited the highly expressed RANKL and OPG in osteoblasts, and obviously decreased the ratio of OPG/RANKL. Conclusions p38MAPK inhibition blocked the differentiation of osteoblasts and decreased the up-regulated OPG and RANKL expressions in osteoblasts significantly (P<0.05).

BACKGROUND:Over-expression of serum interleukin-6 and interleukin-8 may be involved in tourniquet-induced limb ischemia-reperfusion injury to the lung ventilation function. OBJECTIVE:To evaluate the tourniquet effect on serum interleukin-6 and interleukin-8 levels of the rat limb within the safety time limit. METHODS: Sixty rats were randomly divided into control and experimental groups, 30 rats in each group. Rats in the control group had no ischemic preconditioning and were directly subjected to stop bleeding for 2, 3, 4 hours; rats in the experimental group were subjected to ischemic preconditioning (short-time hemostasis for several times within 1 day before ischemia-reperfusion injury), and then underwent 2-, 3-, 4-hour hemostasis at the 2nd day. At 1, 3, 7, 14 days after the recovery of limb blood flow, blood samples were extracted to detect serum interleukin-6 and interleukin-8 levels using ELISA method. RESULTS AND CONCLUSION:The levels of interleukin-6 interleukin-8 showed an increasing and decreasing trend in the two groups, which both reached the peak at the 3rd day (P< 0.05). The levels of interleukin-6 in the experimental group was significantly higher than that in the control group at 1, 3, 7, 14 days (P < 0.05), and only at 7 days, the level of interleukin-6 in the rats undergoing 3-hour hemostasis was significantly lower than that in the control group (P< 0.05). At 7 days, the level of interleukin-8 in the rats undergoing 3-hour hemostasis was lower than that in the control group, but there was no significant difference (P > 0.05), and at 3, 7, 14 days, the level of interleukin-8 in the rats undergoing 4-hour hemostasis was significantly higher than that in the control group (P< 0.05). At 1 and 3 days, the levels of interleukin-6 and interleukin-8 in the experimental group had an increasing trend with the bleeding time and ranked as folows: 2-hour hemostasis < 3-hour hemostasis < 4-hour hemostasis, and there was a significant difference; while in the control group, there was also an increasing trend in the levels of interleukin-6 and interleukin-8, but there was no statistical difference. These findings indicate that the tourniquet preconditioning treatment is preferred at 3 days after limb ischemia-reperfusion injury, when the inflammatory response was the most obvious in rats, and this treatment can dramaticaly reduce inflammatory response. Additionaly, the inflammatory become more obvious with the bleeding time.

0.05).⑥Complications:1 case in the prosthetic disc nucleus replacement group had migration of the device in 1 year after the surgery,but no symptom and did not need revision;1 case complained of bilateral leg pain and 2 cases developed right leg pain in the fusion group,which were treated with oral medications and resolved after the 3-month follow-up.All patients with leg symptoms had surgery at the L5S1 disc space.CONCLUSION:①Quantitative clinical outcomes following prosthetic disc nucleus replacement are at least equivalent to clinical outcomes with traditional lumbar spinal fusion.These results support earlier reports in the literature.②Prosthetic disc nucleus replacement is a safe and effective alternative to fusion for the surgical treatment of symptomatic lumbar disc herniation in properly indicated patients.