Objective To optimize and verify the size exclusion chromatography-high performance liquid chromatography(SEC-HPLC) method for the determination of recombinant human growth hormone(rhGH)-Fc immunofusion protein polymer.Methods The multimer content of rhGH-Fc immunofusion protein was detected by SEC-HPLC.The detection conditions(salt concentration,mobile phase pH,flow rate,column temperature and column model) were optimized to observe the separation effect of the target proteins and polymers.The system suitability,specificity,linearity and range,precision,accuracy and limit of quantification of the method were verified.Results The optimized method was to use TSK-gel G2000SW_(x1)column(5 μm,7.8 mm × 300 mm),mobile phase of 50 mmol/L phosphate buffer(pH 6.80),detection wavelength of280 nm,injection volume of 100 μL,flow rate of 0.6 mL/min and column temperature of 45 ℃.The resolution of rhGHFc immunofusion protein and polymer,the theoretical plate number and the tailing factor all met the requirements;the peak time of rhGH-Fc immunofusion protein was the same as that of the control,while the peak time of GH national standard was different from that of the control,and the protein buffer showed no peak;the concentration of rhGH-Fc immunofusion protein was in the range of 0.307～1.842 mg/mL with good linear correlation between the peak area integral value and the injection volume(R~2=0.999 4);the RSD of peak area and purity in repeatability verification were 0.7% and 0.1%,respectively;the RSD of intermediate precision verification was 0.8%;the average recovery rate of accuracy verification was 99.1% with the RSD of 1.9%;the limit of quantification was 6 μg/mL.Conclusion The optimized SEC-HPLC method was used to detect the content of polymer in rhGH-Fc immunofusion protein with improved accuracy,and the column efficiency and separation were in accordance with the relevant requirements of Chinese Pharmacopoeia(Volume Ⅳ,2020edition),which could be used for the detection of polymer content in samples.

Objective To observe the changes of serum bilirubin(BIL) and high-sensitivity C-reactive protein(hs-CRP) after percutaneous coronary intervention(PCI) in acute myocardial infarction(AMI) patients.Methods The TBiI,IBil,DBil and hs-CRP levels in serum were detected in 60 AMI patients within 6 hours after attack with immunoturbidimetry before PCI,at the point of operation,6h,12h,24h,72h and 7d afer PCI.30 healthy persons were chosen as normal control.The two groups were compared.Results The hs-CRP level was increased gradually with time in AMI patients after PCI.The peak value was at 72h after PCI and it was significantly higher than those in the normal group( P ＜ 0.05 ).The TBil,IBil,DBil levels at pre-PCI point were significantly lower than the normal group (P ＜ 0.05).These index were gradually recovered to the normal group and no significancet differences between them (P ＞ 0.05 ).The coefficient correlation of hs-CRP and TBil,IBil,DBil were 0.44 ( P ＞ 0.05 ).Conclusion The TBil,IBil,DBil and hs-CRP levels in short time after attack of AMI with PCI presented a dynamic changing and recovered to the normal level.No associativity was observed between them,but as the follow-up index,it was significant for the disease turnover.

Objective To explore the sedative effects of a low dose of dexmedetomidine (DEX) during percutaneous kyphoplasty (PKP) in the elderly patients.Methods Forty elderly patients with ASA grade Ⅱ-Ⅲ grade who scheduled PKP were divided into DEX group and control group by random digits table method,each group 20 cases.DEX group received an infusion of 0.5 μg/kg for 10 min before the operation,then the infusion speed was adjusted to 0.2 μ g/ (kg·h).An equivalent volume of 0.9％ sodium chloride was administered in control group.Systolic blood pressure(SBP),diastolic blood pressure(DBP),heart rate (HR),and pulse oxygen saturation (SpO2) were recorded at the beginning of DEX infusion (T0),10 min(T1),30 min(T2),60 min(T3) after DEX infusion and the end of operation (T4),Ramsay scores was also recorded.Results HR,SBP,DBP in DEX group at T1-T3 was significantly lower than that T0 and the same time in control group [(65.6 ± 14.6),(63.4 ± 10.2),(66.7 ± 10.5) times/min vs.(81.4 ± 11.2) times/min and (75.3 ±6.9),(72.8 ± 11.4),(73.8 ±9.6) times/min; (127.5 ± 10.2),(125.4 ± 12.6),(122.4 ±7.8) mm Hg(1 mm Hg =0.133 kPa) vs.(152.5 ± 11.8) mm Hg and(145.1 ± 10.6),(137.2 ± 11.3),(142.6 ± 8.8) mm Hg; (73.7 ± 8.9),(70.3 ± 9.9),(70.3 ± 11.5) mm Hg vs.(90.5 ± 11.0) mm Hg and(86.2 ± 11.6),(82.6 ± 10.9),(78.8 ±8.7) mm Hg,P ＜0.05].The Ramsay scores in DEX group at T1-T3 were significantly higher than those T0 and the same time in control group [(3.6 ± 0.8),(4.2 ± 0.8),(3.7 ± 0.5) scores vs.(2.0 ± 0.0)scores and (1.7 ± 0.6),(1.9 ± 0.4),(1.6 ± 0.5) scores,P ＜ 0.05].HR,SBP,DBP in DEX group at T4 was significantly lower than that T0,HR in DEX group at T4 was significandy lower than that in control group,difference was statistically significant(P ＜ 0.05).Bradycardia and applications vasopressors did not occur in two groups.There was no significant difference in SpO2 between DEX group and control group(P＞ 0.05).Conclusion It is both feasible and safe that sedative effect of a small dose of DEX during PKP in the elderly patients.

One hundred male weanling rats were fed a zinc deficient diet(ZD) containing 7.9ppm Zn for 4 weeks. 56 rats selected as ZD models were divided into 7 groups in random. The rats in 6 test groups were supplied a diet with 40, 60, 80 ppm of zinc enokitake (ZE) and zinc sulfate (ZS) respectively, while the remainder were fed the ZD diet continuously. 5 weeks later, the plasma zinc in 60,80 ppm ZE groups was significantly higher than that in ZS groups in the same doses. The correct rate of learning and memory showed the same trend.Though after that zinc in 60 ppm groups was added to 120 ppm, but the 120 ppm Zn group failed to show greater effect. And the rats in 80ppm ZE group also had the highest level of femur zinc. The RBA of ZE was 207% for plasma and 181% for femur to ZS.The study shows that ZE has a higher biological availability than ZS.

Objective To investigate the mechanisms for cytopathic effect(CPE) of wide-type rubella virus(RV)in ECV304 endothelial-like cells.Method ECV304 cells were cultured in MEM supplemented with 10% fetal calf serum,glutamine,sodium bicarbonate,penicillin and streptomycin at 37℃ with 5% CO-2 in a humidified incubator,and then grown in maintenance medium with 2% fetal calf serum to avoid overstimulating cell growth.Infected cells and control cells were harvested at day 2,4, and 6 after infection for following experiments.RT-PCR and indirect immunofluorescence were used to detect RV infection.Phase-contrast and electron microscopy were performed to analyze morphology changes in infected and uninfected groups.TUNEL assay was carried out to study wide-type RV-induced apoptosis.Result In RV-infected ECV304 cells,cytopathic effects were first observed approximately at 2-3 days post-infection.The majority of cells with CPE changes were of detachment from the monolayer,rounding of cells,cell shrinkage and membrane blebbing were seen.At 4-6 days post-infection,the detached cells increased clearly;the CPE regions on the monolayer were extended,and the infected cells in the CPE regions showed in spindly form with wide spacing.RT-PCR and indirect immunofluorescence results indicated RV infection in ECV304 endothelial-like cells.Using electron microscopy,RV particles and its cytopathic effects including the condensed chromatin,fragmented nuclei and clustering of mitochondria with dramatic changes around nuclear were further examined.TUNEL results showed that the difference of apoptosis index between the infected group and uninfected group was significant(P

Objective To explore the diagnostic value of cystatin C(CysC) on early kidney damage in primary hypertension. Method To detect the CysC, BUN, Scr, UA in Ⅰ (31), Ⅱ (31), Ⅲ (31) stage of primary hypertension patients and 40 control cases ,then compared these results with normal control. Results The level of CysC in serum of Ⅰ , Ⅱ, Ⅲ stage of primary hypertension was(0. 81 ± 0. 16), (0. 94 ± 0. 23), (1.19 ± 0. 46) mmg/L, all of them were higher than normal control (0. 75 ± 0. 10) mg/L. The difference of CysC between Ⅱ、Ⅲ stage of primary hypertension and normal control was significant (t = 4. 195, t = 4. 446, P ＜ 0.01). The level of CysC in Ⅲ stage of primary hypertension was higher than in Ⅰ , Ⅱ stage(t = 4. 382 ,t = 4. 250,P ＜ 0.01). Significant positive correlation was observed between CysC and Scr(r = 0. 713,P＜0.01) ,CysC and UA(r=0. 45,P ＜0.01). Conclusion Serum CysC was a sensitive and reliable indicator on early kidney function damage in primary hypertension.

Objective To evaluate the effects of intravenous analgesia with dezocine combining sufentanil in hip and knee arthroplasty for elderly patients.Methods Ninety patients with ASA Ⅱ ～ Ⅲ and aged ≥ 65 years who were scheduled for hip and knee arthroplasty under CSEA and treated with intravenous analgesia post-operation were selected and divided into three group (20 cases):group Ⅰ (sufentanil 2.5μg/kg + droperidol 2.5 mg),group Ⅱ (sufentanil 2.0 μ g/kg + dezocine 10 mg + droperidol 2.5 mg) and group Ⅲ (intramuscular dezocine 10 mg and sufentanil 2.0 μ g/kg + droperidol 2.5 mg),which were diluted to be 100 ml by adding saline.VSA,Ramsay score as well as side effects were recorded at 6,12 and 24 h after operation.75 mg meperidine was administered intramuscularly as rescue medication when VAS was above 5.Results Compared with group Ⅰ and Ⅲ,group Ⅱ showed better outcomes regarding to the VAS and less side effects.Ramsay score 6 h after operation and the amount of patients who needed meperidine in group Ⅲ were higher than those in the other two groups (P ＜ 0.05).Conclusion Dezocine combining with sufentanil in elderly patients underwent low limb arthroplasty shows better postoperative analgesia effects with less side effects,and it's worth for clinical application.

OBJECTIVE:To develop the method for the determination of riluzole concentration in blood plasma.METH?ODS:Riluzole was extracted with diethyl ether.The residues were analyzed with a reverse phase HPLC system(Diamonsil TM C 18 column,250mm?4.6mm,5?m),with mobile phase of MeOH-0.03mol/L and NH 4 H 2 PO 4 (80∶20,V/V),UV detection wave length of265nm,flow rate of0.8ml/min.RESULTS:The linear coverage of Riluzole was5～1000ng/ml;the lowest detectable concentration was5ng/ml.The average recoveries of riluzole were99.51%,95.74%and97.12respectively.The within-day and between-day relative standard deviations were1.17%、6.48%(n=5)respectively.CONCLUSION:The met_ hod is sensitive,accurate,simple and reliable,and it can be applied to phamacokinetic studies.

AIM: To study the effects of Huoxue Huayu and Yiqi Wenyang compound prescription (composition; ginseng, radix astragali, plantaginis semen, lignum sappan, chuanxiong, salvia miltiorrhiza, radix aconiti lateralis preparata, bitter orange, cassia twig) of TCM on the congestive heart failure in rats. METHODS:The rats were given doxorubicin hydrochloride by intraperitoneal injection to establish the model of congestive heart failure and were randomly divided into control group, congestive heart failure model group, Xinbao treatment group ( XB treatment) and Huoxue Huayu and Yiqi Wenyang compound prescription treatment group (QXYTM treatment). Normal saline, liquid medicine Xinbao and liquid medicine Huoxue Huayu and Yiqi Wenyang compound prescription were administered respectively to four groups by gavage. Radioimmunoassay, semi - quantitative RT - PCR and flow cytometry methods were used to determine the expressions of plasma atrial natriuretic peptide ( ANP) , renal medulla aquaporin - 2 ( AQP2 ) and heat shock protein70 (HSP70) in congestive heart failure rats before and after treatment. RESULTS: In congestive heart failure rats treated with Huoxue Huayu and Yiqi Wenyang compound prescription, the cardiac output and left ventricle systolic pressure increased significantly, the left ventricular end diastolic pressure decreased, and the maximum ascending and declining rate of left ventricular pressure ameliorated, thus the hemodynamics was effectively improved. Meanwhile, the level of plasma cardionatrin decreased, the abnormal expression of renal medulla AQP2 was markedly restored, and the transcription and expression of HSP70 gene were also increased. CONCLUSION: Huoxue Huayu and Yiqi Wenyang compound prescription promotes the recovery of the pathogenetic conditions of congestive heart failure in rare. The mechanism may be in association with a decrease in plasma ANP level, correction of renal medulla AQP2 abnormal expression and enhancement of HSP70 expression.

Objective To investigate the clinical significance and relationship of preS1, HBV DNA and HBV-M. Methods PreS1 and HBV-M was detected by ELISA method,and HBV DNA was detected by PCR. Then the results were analyzed. Results In HBV patients,the positive rates of preS1 and HBV DNA had no statistically significant ,they had fine dependability. The detection rate of preS1 in HBeAg(+) group(80.3%) and HBeAg(+)group( 56.3% ) had statistically significant. In some patients,though HBeAg had become negative, HBV still replicated. In HBV DNA replicated patients(≥103 copies/ml) ,the detection rate of HBeAg and preS1 were 51.5% and 70.9% ,they had statistically significant. Conclusion HBV DNA and PreS1 had fine dependability,preSl could reflect the replication of HBV sensitively than HBeAg,it could be used as a reliable new marker of HBV replication in vivo.