Deafness ; Diabetes Complications ; diagnosis ; epidemiology ; Diabetes Mellitus ; epidemiology ; Hearing Loss ; diagnosis ; etiology ; Humans

Objective To explore the respective advantages of different direction Hegu needlings by using them to treat the pain stage of scapulohumeral periarthritis (periarthritis of shoulder) and investigating their clinical therapeutic effects. Method A randomized controlled trial was carried out. Sixty patients with scapulohumeral periarthritis in the pain stage were allocated into group Ⅰ (30 cases) and group Ⅱ (30 cases). Group Ⅰ received acupuncture at points Jianyu(LI15), Jianliao(TE14) and Jianzhen(SI9) in the direction parallel to the meridian course and group Ⅱ , in the direction perpendicular to the meridian course. Treatment was given 30 min once for a total of 12 times. The simplified Mcgill Pain Questionnaire score and the Japanese Orthopaedic Association (JOA) score were recorded before and after treatment. Result There were no statistically significant differences in the pain, joint activity and joint function between the two groups before, during and after treatment. Conclusion Two different direction Hegu needlings both have a marked therapeutic effect on the pain stage of scapulohumeral periarthritis. The two have no significant difference.

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

Objective To investigate the protective effect of Ambroxol against experimental lung ischemia reperfusion (I/R) injury in a situ hilar clamp model.Method Left lung of rat was rendered and ischemic for 90 minutes,and reperfused for up to 2 hours,as the model.Twenty-four SD rats were randomly divided into 3 groups with 8 rats each group:control group,I/R group,I/R and Ambroxol treatment group (AMB group).Rats of AMB group received Ambroxol (25 mg/kg) Intraabdominally 30 minutes before ischemia and intravenously 5 minutes before reperfusion.After 2 hours of reperfusion,blood-gas analysis,the serum level of IL-1?,IL-8 and TNF-?from carotid artery were delected.The wet/dry ratio of lung,the activity of erythrocuprein (SOD),the content of malonaldehyde (MDA) and the activity of myeloperoxidase (MPO) were determined and pathematology changes were observed in the left lung tissue.Differences within the groups were analysed using two-sample t-test. Results After 2 hours of repeffusion,there were no significant changes of artery partial pressure of oxygen (PO_2) and partial pressure of carbon dioxide (PCO_2) among three groups.The wet/dry ratio of lung,the activity of MPO (U/g) and the content of MDA (nmol/mgprot) of I/R group were (5.3?0.5),(1.30?0.26) and (0.66?0.16),significantly higher than those of the control group (P

OBJECTIVE:To analyze the utilization of different dosage forms of essential drugs in the clinical practice in our sanatorium and the tendency of drug use of patients.METHODS:The utilization of essential drug in our sanatorium were analyzed statistically in respect of drug category,constitution ratio of drug use.A questionnaire survey was carried out to survey the tendency of drug use of patients.RESULTS:Tablet was the most common dosage form in the clinic.Antibiotics and drugs for clinical therapy had complete dosage forms.The common dosage forms used patients were tablets and capsule,etc.CONCLUSIONS:Patients of our sanatorium select drugs tendentiously.The tendentious selection of drug is no difference in each age group.

Objective To investigate the prevalence and correlation factors of cardiovascular damage in patients with diabetic nephropathy (DN) and non-diabetic nephropathy (NDN).Methods A total of 278 chronic kidney disease (CKD) patients admitted to the First Affiliated Hospital of Jinan University from January 2014 to May 2016 were enrolled,including 78 case of DN and 200 case of NDN.Patients had cardiac and carotid ultrasonography test by colour doppler ultrasonography,and their clinical and biochemical data were collected.Multiple linear regression analysis and multivariable logistic regression analysis were applied to study the correlation factors of cardiovascular damage in CKD patients.Results Mean age was 48.22 years in the 278-patient cohort,which included 178(64.03％) men.Compared with NDN group,DN patients had higher left atrial dimension,interventricular septal thickness,left ventricular end-diastolic dimension,left ventricular posterior wall thickness,left ventricular mass index (LVMI),carotid intima-media thickness (cIMT) and carotid plaques ratio.Their estimated glomerular filtration rate (eGFR) and the ratio between the peak speed of the early filling wave and that of the atrial contraction wave (E/A ratio) were however lower (all P ＜ 0.05).Prevalence of left ventricular hypertrophy (LVH),left ventricular relaxant function reduction and cIMT thickening in DN group were 67.95％,70.27％ and 57.14％,higher than those in NDN group (40.00％,42.31％ and 17.39％,respectively) (all P＜ 0.05).Along with the progress of CKD,LVMI and LVH proportion in patients with DN and NDN increased gradually.LVMI and LVH proportion in DN patients in CKD 1-2 phase and CKD 3-4 phase were higher than those in NDN patients (all P ＜ 0.05).In all CKD phases,cIMT and cIMT thickening proportion in DN group were higher than those in NDN group (all P ＜ 0.05).Just in CKD 1-2 phase,DN group had lower E/A ratio and higher proportion of left ventricular relaxant function reduction than NDN group (all P ＜ 0.05).After multiple linear regression analysis,gender,BMI,hemoglobin,eGFR and DN were related with LVMI;age,serum calcium and DN were related with E/A ratio;age and DN were related with cIMT (all P ＜ 0.05).In multivariate logistic regression,DN,hemoglobin and eGFR decrease were independently associated with LVH;age and BMI were independently associated with reduction of left ventricular relaxant function;age and DN were independently associated with cIMT thickening in all CKD patients (all P ＜ 0.05).Conclusions DN patients have more severe cardiovascular damage than NDN patients,and DN may be associated with LVMI,E/A ratio,cIMT,LVH and cIMT thickening in all CKD patients.

Objective To express and purify mammalian glycosylation modified trimeric Ebola virus trimeric glycoprotein (EBOV-GP) using novel glycoengineered Pichia pastoris.Methods The EBOV-GP and EBOV-GPΔMLDΔTM genes were cloned into the pPICZ-αA vector,electrochemically converted to glycoengineered Pichia pastoris,and compared with EBOV-GP expressed in HEK-293T cells.Glycosylation was analyzed by PNGaseF and EndoH digestion,while the target protein was purified by affinity chromatography and ion exchange chromatography.N-terminal sequencing was used to determine whether the signal peptide was correctly cleaved during protein translation and gel column analysis was used to find out whether the trimeric structure was formed.Results The results of PNGaseF showed that EBOV-GP expressed by glycoengineered Pichia pastoris and HEK-293T cells had the same relative molecular mass and N-glycosylation degree.EndoH digestion showed that the N-glycosylation modification of EBOV-GPΔMLDΔTM was in a non-high mannose form.N-terminal sequencing showed that the signal peptide of the GP protein itself was correctly excised.Gel column analysis showed that the purified protein was in a trimeric form.Conclusion An EBOV-GP is obtained with complex glycosylation modification based on Glycoengineered Pichia pastoris.

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

In order to solve the problems of erratic drug absorption and low bioavailability after oral administration for poorly-water soluble drugs due to low solubility, a series of novel pharmaceutical dosage forms as solid dispersion, liposome, microemulsion, vesicle, cyclodextrin inclusion complexes and drug nanocrystal have been developed in recent years. Among which drug nanocrystal attracts more attentions for its simpler preparation method, higher drug loading and easier manufacturing technology in the design of dosage forms suitable for different administration routes. In this paper, the nanocrystals of the poorly-water soluble drugs prepared based on bottom-up and top-down technologies were introduced. The characteristics and applications of the nanocrystal-based dosage forms as suspension, tablet and capsule were also introduced and carefully evaluated with the focus on their pharmacokinetics, pharmacodynamics and tissue targeted drug distribution after delivery by oral administration, intravenous injection and pulmonary inhalation. The advantages of drug nanocrystals in their therapeutics effects over the bulk drugs were discussed together with the inherent mechanism. Finally, the problems existing in basic research and scaled-up manufacture of drug nanocrystal as well as the possible ways of solution were listed out so as to make the nanocrystal-based preparations exert their maximum therapeutic effect after clinical application.

Objective To investigate the mechanism and significance of low concentration nitric oxide (NO) inhalation in the treatment of pulmonary thromboembelism in swine. Method The pulmonary thromboem-bolism(PTE) model was made in 15 healthy infantile swines which were subsequently assigned to either control group (n = 8) or NO group (n = 7). Swines of the control group were not treated with any medicine, while 10 ppm of NO was administered by continuous inhalation for 2 hours to swines of NO group. Volume of physiological dead space (VDphy), volume of alveolar dead space (VDalv), intrapulmonary shunt (Qs/Qt), mean pulmonary arterial pressure (PAP), systolic blood pressure (SBP), heart rate (HR), cardiac output (CO), arterial blood pH (pH), arterial partial pressure of carbon dioxide (PaCO2) and arterial partial pressure of oxygen (PaO2) were measured 30 min before and 0 min, 30 min, 60 min, 120 min and 180 min after establishment of VIE. Results The post-FIE VDphy, VDalv, Qs/Qt and PAP in both groups increased markedly after PTE compared with the cor-responding pre-PTE measurements (P < 0.01). Post-FIE PaO2 of both groups decreased significandy (P <0.05 and P <0.01), while significance difference was found between pre- and post-PTE HR, SBP, CO, pH or PaCO2 in neither groups (P > 0.05). Both post-PTE PAP and VDalv in NO group were markedly lower(P <0.05 and P <0.01) and beth PaCO2 and PaO2 were much higher than those of the control group (P <0.05). No signi-fieant difference were found in other measurements between two groups. Conclusions Pulmonary arterial pressure may be lowered, alveoli dead space may be reduced and PaCO2 increased by low concentration NO inhalation for the treatment of PIE without decline in haemodynamic status.