In order to improve the expression level of segment of GABAA receptor a1 subunit in E. coli, growth conditions of the recombatant, which influence the final yield of protein expression, including growth medium, inoculation ratio, temperature, pH, rotation speed, inducing time and concentration of IPTG and so on, were studied in shaking flasks. The results indicated that, with 3% inoculation ratio, cultured 3.5 hours at 37℃, and then induced 5 hours by IPTG at 32℃, the yield of GABAA receptor protein was 95mg/L and the biomass was 3.25 g/ L. In contrast, using a 16 L stirred fermentor instead of shaking flasks, the highest level of the protein expression, 136mg/L with 4.95g/L of biomass, was achieved after fermenting 5.5 hours.

Objective To observe the ultrastructure changes of detrusor of urinary bladder following partial outlet obstruction. Methods 14 New Zealand white male rabbits were divided into "A" and "B" group.The "A" group rabbits underwent operation to induce bladder outlet obstruction,"B" group rabbits served as controls.The ultrastructures including rough endoplactmic reticulum and mitochondrion of detrusor cell of each bladder were examined after 5 weeks. Results The average area of rough endoplactmic reticulum in the obstructed group was (5.377?2.318) ?m 2 in each unit cytoplasm area (115.28 ?m 2) versus (0.476?0.319) ?m 2 in the controls ( P

Objective To observe the effect of electroacupuncture on improvement of learning and memory ability and the expression of connective tissue growth factor(CTGF)mRNA and protein in hippocampus in diabetic rats with cognitive impairment.Methods The rat diabetes model was induced by injecting streptozotocin(20 g/L),and then the rats were randomly divided into three groups:electro-acupuncture group(EA),diabetes-mellitus-untreated group(DM)and control group(CN).After four weeks of electroacupuncture treatment,blood glucose level was determined and the effect of electroacupuncture on learning and memory was examined with the device of Morris water maze.RT-PCR was used to detect CTGF mRNA level,and immunohistochemistry was used to detect CTGF protein expression.Results Blood glucose level and the latency period in DM group were increased compared with those in EA and CN groups(P

Blood Coagulation ; Blood Viscosity ; China ; Diagnosis, Differential ; Humans ; Medicine, Chinese Traditional

Blood Coagulation ; Blood Viscosity ; Diagnosis, Differential ; Humans ; International Cooperation ; Medicine, Chinese Traditional

OBJECTIVE To investigate the immunoregulatory role of tolerogenic dendritic cells(TolDC) in allergic airway inflammation in mouse model of allergic rhinitis(AR).METHODS A total of 24 Balb/c mice were equally and randomly divided into 4 groups with 6 mice in each group. AR group was established with oval bumin(OVA) sensitization and challenge, meanwhile the physiological saline(PBS) sensitization and challenge as the control group. AR group were treated by adoptive transfer of TolDC as treatment group. And treatment group were injected intraperitoneally with TGF-β/IL-10R neutralizing antibody as blockade group. In each group, allergic nasal symptoms score, inflammatory cell infiltration in lung, the expression of Th1/Th2-derived cytokines in the bronchoalveolar lavage fluid(BALF) and nasal lavage fluid(NALF), the expression of OVA-specific serum IgE and the expression of Treg in lung were measured.RESULTS The mouse model of OVA-induced AR was successfully developed. Compared with AR group, treatment group exhibited lower allergic nasal symptoms score, a decrease in inflammatory cell infiltration in lung, lower expression of Th1/Th2-derived cytokines in BALF and NALF and OVA-specific serum IgE, as well as up-regulation of Treg in lung, which were abolished by TGF-β/IL-10 neutralizing antibody shown in blockade group.CONCLUSION TolDC suppress airway inflammation in AR by inducing regulatory T cells through TGF-β/IL-10-dependent mechanisms.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

Objective To investigate the role and potential mechanism of interleukin-17A (IL-17A) in the inflammatory response to traumatic brain injury (TBI) in rats.Methods The adult male Wistar rats were randomly (random number) divided into seven groups:control group (n =6),sham operation group (n =6),TBI group (n =24),sham operation + normal saline group (n =6),sham operation + Y320 (an immunomodulator acts as an inhibitor of IL-17A) group (n =6),TBI + normal saline group (n =6) and TBI + Y320 group (n =6).The TBI model of rat was established by using free-falling-body impact device.The levels of IL-17A and nuclear transcription factor kappa B p65 (NF-κB p65) in the cerebral cortex were assayed by using Western Blot.The capability of leaming and memory of rats was assessed by Morris water maze.The beam balance test was employed to evaluating the neurological motor performance and the capability of balance.Results Compared with the sham operation group,the levels of IL-17A and NF-κB p65 in the cerebral cortex of TBI,TBI + saline and TBI + Y320 groups increased significantly (P ＜0.05) and peaked at the 3rd day after TBI.Compared with TBI + normal saline group,the level of NF-κB p65 was significantly down regulated by Y-320 (P ＜ 0.05) at the 3rd day after TBI in TBI + Y320 group.The lengths of latency time required for rats to escape to the platform area in TBI + normal saline group were (57.72±3.29) s,(55.63±3.85) s,and (55.02±3.92) sat the3rd,5th and7th days after TBI,respectively;while those in TBI + Y320 group were (35.45 ± 3.04) s,(30.98 ± 2.92) s,and (23.90 ±2.51) s at the 3rd,5th and 7th days after TBI,respectively.Thus,the capability of learning and memory of rats in TBI + Y320 group was improved significantly 3d,5d and 7 days after TBI (all P ＜ 0.01).Conclusions This study shows IL-17A is involved in the process of secondary brain injury after TBI,and associated with inflammation by activating the NF-κB p65 signaling pathway.

Calcinosis ; Fibula ; Free Tissue Flaps ; pathology ; Humans ; Reconstructive Surgical Procedures

Objective To evaluate the application of microvascular coupler device (MVCD) in the free-tissue flap transfer for head-and-neck defects reconstruction.Methods Its a retrospective study of the clinical data of 743 patients who received 763 free-tissue flap transfer for head and neck defects reconstruction in Hunan Cancer Hospital from January,2014 to January,2016.For microvascular anastomoses,413 were done manually (the manual group) and 350 done with MVCD (the MVCD group),of which the latter included end-to-end venous anastomosis in 159,endto-side venous anastomosis in 190 and end-to-end arterial anastomosis in 1.The time for anastomosis,rate of blood leakage from the anastomosis and venous thrombosis,and the survival rate of flaps were compared between the manual group and the MVCD group.The impacts of types of anastomosis of end-to-end vs end-to-side (both with MVCD),on the formation of venous thrombosis and survival of flaps were also recorded.The analysis was performed under t-test and chi-square test using SPSS software 19.0 with P ＜ 0.05 for differences with a statistical significance.Results Time for anastomosis was significantly shorter in the MVCD group (4.43±0.51min) than in the manual group (14.75± 2.43min,P＜0.05).The rates of anastomosis leakage were 0 in the MVCD group and 9.7％ in the manual group(P＜0.01),flap necrosis were 0.57％ and 1.69％ (P=0.15),the rate of venous thrombosis were 1.15％ and 3.63％(P=0.02),respectively.There was no significant difference in the time for anastomosis,the rate of venous thrombosis and the survival rate of flaps with end-to-end anastomosis and end-to-side anastomosis with MVCD.Conclusion The application of microvascular coupler device is valuable in the reconstruction of head-and-neck defects with free-tissue flaps because it can significantly shorten the time for anastomosis,decrease the ocurrence of anastomosis leakage and venous thrombosis,reduce the risk of flap failure.