Objective To investigate the expression of aquaporin-4 (AQP4) in contused brain and contralateral side and correlation of AQP4 level with brain edema.Methods A total of 70 health adult SD rats were divided into sham operation group (n =10) and brain contusion group (n =60) according to the random number table.Except for no impact in sham operation group,the remaining operations were the same in both groups.Ten rats in brain contusion group were separately sacrificed at 1,6,24,48 and 72 hours,and 7 days.Pathological changes in brain tissues of rats were detected and immunohistochemistry staining and Western-blot were used to measure AQP-4 expression.Results There were no abnormalities of brain tissue structure in sham operation group with AQP4 expressed mainly in glial cells and vascular endothelial cells.With respect to the injured side in brain contusion group,mixed brain edema that consisted of angioedema and cellular edema occurred along with tissue necrosis,massive inflammatory cell infiltration,and microgial proliferation.AQP4 was apparently expressed in gliocytes and vascular endothelial cells,which showed a fall at 1 hour,the lowest at 6 hours,the peak at 24 hours,slow fall at 48 hours,a second peak at 72 hours,and then a fall until 7 days.Level of AQP4 other than that at 1 hour had significant difference compared with sham operation group (P ＜ 0.01).Whereas the contralateral side in brain contusion group showed no pathological abnormalities at 1 hour,but cellular edema emerged at 6 hours followed by mixed brain edema mainly characterized as cellular edema at 24 hours,continued cellular edema but alleviated angioedema at 48 and 72 hours,and alleviated cellular edema at day 7.AQP4 was up-regulated in gliocytes and vascular endothelial cells,which decreased at 1 hour (1.313 ± 0.01),minimized at 6 hours (0.922 ± 0.03),peaked at 24 hours (2.848 ± 0.020),fell at 48 hours,peaked again at 72 hours (2.662 ± 0.02) and fell to almost normal level at day 7.By contrast with sham operation group,level of AQP4 had significant difference other than that at 1 hour and 7 days (P ＜ 0.01).Conclusions After brain contusion,angioedema and subsequent cellular edema emerge in the contused side.Pathological changes are delayed in non-contused side with cellular edema ahead of angioedema.Level of AQP4 is closely related with traumatic brain edema.

Objective To explore the effect of Buyanghuanwu decoction on blood glucose control and renal function protection in patients with early diabetic nephropathy.Methods 170 cases of patients with early diabetic nephropathy from May 2012 to June 2013 in wenzhou hospital of integrated traditional chinese and western medicine were selected,all patients were divided into observation group and control group,the control group was given routine treatment,the observation group treatment with Buyanghuanwu decoction based on control group .The clinical effective rate was compared between the two groups after treatment,the levels of blood lipids, 24 h urine microalbumin and hemorheology were compared between the two groups before and after treatment.Results After treatment, the total effective rate of the observation group was 90.59%,which was significantly higher than the control group 76.47% (P<0.05).Compared with before treatment,the levels of triglycerides and cholesterol decreased in two groups,high-density lipoprotein levels increased,24 h urine microalbumin decreased,whole blood viscosity,whole blood viscosity trimming and fibrinogen decreased,the difference was statistically significant(P<0.05).Compared with the control group,the levels of triglycerides and cholesterol in observation group was lower,high-density lipoprotein was higher,24 h urine microalbumin was lower,whole blood viscosity,whole blood viscosity trimming and fibrinogen were lower,the difference was statistically significant(P<0.05).Conclusion Buyanghuanwu decoction in the treatment of diabetic nephropathy is effective in treating diabetic nephropathy,it can effectively reduce blood lipids and blood viscosity,significantly reduce urinary protein excretion and protect the kidneys.

Objective To explore advantage of RNA guiding activity in vitro cleavage assay and intracellular antivi-ral effects for hepatitis C, basing on one external guide sequence (EGS)sequence of the conserved region of 5′UTR of HCV genome .Methods We analyzed the sequence and structure of the conserved region of 5′UTR of HCV ge-nome with computer software and designed one EGS which was direct at the potential targeting site ( C67-G68 ) .We tested the gene expression of HCV in Northern blot and Western blot and its antiviral activity by fluorescence quan -titative PCR in the Huh7.5.1 cell which was transfected by EGS-C67 .Results EGS-C67 had obvious advantage of RNA guiding activity in vitro cleavage assay .EGS-C67 obviously inhibited the gene expression of HCV and inhibit the proliferation of HCV and down-regulate the titer of HCV about 150 times( P<0.01 ) .Conclusions EGS-C67 , the new antiviral RNA has notably antiviral activity in cultured cells , thus could provide a new promising approach for development of anti-HCV strategy .

Through the study on traditional Chinese medicine (TCM) data mining methods, the traditional mining model was revealed to have certain limitations. New ways and means should be searched in order to tap the potential, deep level and greater value of TCM knowledge. In view of the situation, advantages of strategy pattern thought for medicine data mining were discussed. And deep research was carried on. New ideas on TCM data mining based on strategy pattern was proposed.

Hepatitis C virus (HCV), one of the major pathogens of viral hepatitis, causes significant hazards in humans. Interferon treatment in combination with ribavirin is used as the first line clinical treatment for HCV infection. However, good response to this treatment has only been observed in few patients and repeated recurrence has also been reported frequently. Therefore, new antiviral agents and therapies are in urgent demand. Here, we report a newly constructed Escherichia coli RNase P based M1GS ribozyme that can specifically and efficiently target the core gene of HCV. The guide sequence (GS) of this M1IGS was designed according to the sequence of the core coding region of HCV genome. The GS was then covalently linked to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The specification of this sequence-specific ribozyme, M1GS, was then examined using an in vitro cleavage assay. The cytotoxicity and its activity in inhibition of HCV gene expression and viral proliferation were further studied in vivo. Our results show that the reconstructed M1GS ribozyme displayed obvious catalytic activity in cleaving target mRNAs fragment in vitro. Notable reduction in the expression of HCV core protein and a 1 000-fold reduction in viral growth were also observed in cultured HCV infected Huh7.5.1 cells expressing the functional M1GS ribozyme. This study demonstrated a direct evidence for the antiviral activity of the customized M1GS-HCV/C141 ribozyme, and thus provided a promising new strategy for clinical treatment of HCV infection.
Antiviral Agents ; pharmacology ; Escherichia coli ; genetics ; Genetic Engineering ; Hepacivirus ; genetics ; physiology ; RNA, Catalytic ; genetics ; pharmacology ; RNA, Guide ; genetics ; Ribonuclease P ; genetics ; Viral Core Proteins ; genetics

Objective To compare the characteristics of plantar pressure between old and young people walking on flat. Methods 30 persons aged 55-65 years and 30 students aged 18-23 years were measured with a kind of plantar pressure measurement Footscan when they walked naturally, including impulse, max force, load rate of 10 regions, and distribution of the contact area of medial and lateral heels. Results The impulse was significantly different in 7 regions between both sides in the old people, and in 3 regions in the students. The impulse was significantly less in the old people than in the students (P<0.05). The max force was significantly different in 6 regions between both sides in the old people, and in 3 regions in the students. The max force was significantly less in the old people than in the students (P<0.05). The load rate was significantly different in 7 regions between both sides in old people, and in 3 regions in students. The load rate was significantly less in the old people than in the students (P<0.05). The contact area was larger in the medial heel than the lateral heel in old people group (P<0.05), and it was different between left and right foot. While the contact area was smaller in the medial heel than the lateral heel in the students (P<0.05), and there was no difference between left and right foot (P>0.05). Conclusion The impulse, max force, load rate decrease in the old people with the imbalance of distribution of the pressure between left and right.

Better governance of private hospitals is key to deepening the health reform.In view of the present development of private hospitals in Hubei province,this paper expounded the characteristics development of private hospitals and the governance of specific measures in the province and analyzed the defects found in the legal system and mechanism.The authors also raised such recommendations as perfecting the legal system and improving the mechanism of information disclosure and evaluation.

Objective To investigate the therapeutic effect of eperisone on patients with subacute spinal cord neuropathy with incomplete spasm.Methods Forty-eight patients with Subacute spinal cord neurosis combined degeneration of both lower extremity incomplete spasm in Department of Neurology,Zhejiang Quhua Hospital were selected and randomly divided into control group and study group.The control group were treated with folic acid and mecobalamin,the study group received more with eperisone,and 14days for one course,two groups were treated for two course.Ashworth spasm Scale,simple Fugl-Meyer score scale,improved Barthel index,bilateral H tibial nerve reflex status,clinical efficacy and complications were compared before and after the treatment.Results After treatment,levels of the flexion and extension of knee joint in 2 groups increased,and the scores of Ashworth spasticity scale decreased,scores of FMA and MBI increased,the latency of H reflex latency increased,levels of Hmax/Mmax decreased(P<0.05),compared with the control group,levels of the flexion and extension of knee joint in the study group were higher,and the scores of Ashworth spasticity scale were lower,scores of FMA and MBI were higher,the latency of H reflex latency were higher,levels of Hmax/Mmax were lower(P<0.05),the effective rate of the control group(66.67％)was lower than the study group(91.67％)(P<0.05),the non-performing rate in 2 groups had no difference(25.00％Vs 20.83％).Conclusion Eperisone has definite effect on patients with subacute spinal cord neuropathy with incomplete spasms.it can decrease muscle tension and spasticity and improve neurological function.

Objective The establishment of the model of myocardial ischemia-reperfusion injury (MIRI) in SD rats involves lengthy time, severe damage, and death of the rats.We aimed at an efficient and rapid method for establishing an MIRI model in rats to ensure a high success rate of modeling and survival of the animals.Methods Using the trachea intubation-assisted breathing machine, we fully exposed the hearts of 80 adult SD rats (8-10 weeks old and weighing 250-300 mg) by mutilating the 2-3 intercostal muscles.Then, we rapidly positioned and ligated the left anterior descending (LAD) coronary artery for an hour, and established the model of MIRI at 48 hours after releasing the slipknot.We performed electrocardiography (ECG) before, during, and at 1 and 48 hours after operation, ligated the same part again at 48 hours postoperatively, and measured the size of the myocardial ischemia and infarction areas using Evans-TTC double staining.Results The survival rate of the 80 rats was 90% and the success rate of MIRI modeling was 86.25%.After ligation of the LAD coronary artery, ECG showed the ST segment and T wave elevation, followed by gradual decrease of the ST segment and R wave voltage, the myocardium and cardiac apex muscle grey or dark grey below the ligation slipknot.The heart rate myocardial motion amplitude were obviously reduced after ligation.Evans-TTC double staining revealed an evident myocardial infarction area at 48 hours after modeling and the ratio of the ischemia area + infarction area to the total myocardial area of the left ventricle was 0.43 to 0.55 (P>0.05) and that of the infarction area to the ischemia area + infarction area was 0.35 to 0.45 (P>0.05).Conclusion The modified rat model of MIRI can be fast and efficiently established, with a high success rate of modeling and a low mortality of the animals.

Objective To compare the effects of celecoxib and amniotic membrane suspension (AMS) on corneal neovascularization (CNV) area and expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloprotein-ase-9 (MMP-9) in the growth of corneal neovascularization after thermal burn in rabbits,and provide a theoretical basis of celecoxib for the clinical treatment of corneal neovascularization.Methods Left corneas of 36 rabbits were burned by the home-made burning-device,and randomly divided to three groups:negative control group (n =12),AMS group (n =12) and celecoxib group (n =12),were respectively sub-conjunctival injected by 90 g · L-1 saline (0.1 mL),AMS (0.1 mL) and 8 mg · mL-1 celecoxib solution (0.1 mL).The histological morphology,growth condition and area of CNV were compared under slit lamp microscope at 4 days,7 days and 14 days after thermal-burned.At 7 days after thermal-burned,four appropriate corneas were randomly taken to detect the expression of MMP-2 and MMP-9 by immunohistochemistry,and the results were analyzed by computer image analysis system.Results At 4 days,7 days,14 days after thermal-burned,the areas of neovascularization were (11.32 ± 1.11)mm2,(38.49 ± 4.64) mm2,(43.30 ± 4.39) mm2 in negative control group,(9.69 ± 1.30) mm2,(31.15 ± 4.85)mm2,(37.19 ± 5.27) mm2 in AMS group,(8.47 ± 1.20)mm2,(30.31 ± 4.93) mm2,(36.69 ± 3.54) mm2 in celecoxib group,respectively.At different time points,neovascularization area in AMS group or celecoxib group was significantly lower than negative control group (all P ＜ 0.05).There was no difference between AMS and celecoxib group (all P ＞ 0.05).At 7 days after thermal-burned,the expression of MMP-2 and MMP-9 was not different between AMS group and celecoxib group (all P ＞ 0.05),and significantly lower than negative control group (all P ＜ 0.05).Conclusion Celecoxib and amniotic membrane suspension can all effectively inhibit CNV after thermal-burned,which may be related to the down-regulated expression of MMP-2,MMP-9 in thermal-burned corneas.