OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets， followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established ， and the zebrafish were treated with L. ruthenicum Murr. extract （LRME） at concentrations of 0.1， 0.2 and 0.4 mg/mL， respectively； 24 h later， behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 （IL-6）， IL-1β， IL-10， tumor necrosis factor-α （TNF-α）]， oxidative stress markers [superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， catalase （CAT）]， and neurotransmitters [5- hydroxytryptamine （5-HT）， γ-aminobutyric acid （GABA）， glutamic acid （Glu）， dopamine （DA）， and norepinephrine （NE）] were measured. The protein expression levels of protein kinase B1 （AKT1）， phosphorylated AKT1 （p-AKT1）， epidermal growth factor receptor （EGFR）， B-cell lymphoma 2 （Bcl-2）， sarcoma proto-oncogene，non-receptor tyrosine kinase （SRC）， and heat shock protein 90α family class A member 1 （HSP90AA1） in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these， apigenin， naringenin and others were recognized as core active compounds， while AKT1， EGFR and others served as key targets； EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-， medium-， and high-dose groups were 54.60%， 69.03% and 77.97%， 开发。E-mail：hjp_yft@163.com respectively， while the inhibition ratios of locomotor distance were 0.57， 0.83 and 0.95， respectively. Compared with the model group， the number of resting counts， resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups （P＜0.05）. Neuronal damage in the brain was alleviated. Additionally， the levels of IL-6， IL-1β， TNF-α， MDA， Glu， DA and NE， as well as the protein expression levels of AKT1， p-AKT1， EGFR， SRC and HSP90AA1， were markedly reduced （P＜0.05）， while the levels of IL-10， SOD， GSH-Px， CAT， 5-HT and GABA， as well as Bcl-2 protein expression， were significantly elevated （P＜0.05）. CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects， and its specific mechanism may be related to the regulation of inflammatory responses， oxidative stress， neurotransmitter balance， and the EGFR tyrosine kinase inhibitor resistance signaling pathway.

Objective To investigate the effects of harmine(HM)on the expression level of mitochondrion fusion related proteins and mitochondrial function injury in PC 12 cells.Methods PC 12 cells were divided into cell control group,HM group,mitochondrion mitosis inhibitor Mdivi-1 group,HM+Mdivi-1 group,mitochondrion fission agonist WY14643 group,HM+WY14643 group,with drug concentrations of 1,10,25,50,100 μmol·L-1.After 24 h treatment,the MTT method was used to detect the cell survival rate,and a microscope was used to observe the cell morphology,MitoTracker Red probe staining was used to observe the mitochondrial morphology and the length ratio of vertical and horizontal axes,JC-1 staining was used to detect the mitochondrial membrane potential,and a kit was used to detect ATP level and lactate dehydrogenase(LDH)activity.Immunofluorescence staining and Western blotting were used to assess the expression levels of caspase-3,apoptosis-promoting protein(Bax)cytochrome C(cyt-c),mitochondrial fusion protein(Mfn2)and mitochondrial mitotic protein(Drp-1).The interference sequence of Drp1 was transfected by electroporation,and the siRNA sequence with good transfection effect was screened.The related indicators were detected by fluorescence method,MTT method,and immunoblotting method in cooperation with drug intervention.Results MTT results showed that compared with the cell control group,the survival rate of HM group,Mdivi-1 group,HM+Mdivi-1 group,WY14643 group and HM+WY14643 group decreased significantly(P＜0.01),and the EC50 were(11.48±2.32),(12.35±1.67),(14.88±2.07),(39.14±3.25),(20.09±1.97),respectively.According to this,subsequent experiments selected 20 μmol·L-1for HM,WY 14643 and HM+WY14643 as working concentrations to construct PC 12 cell model.Microscopic observation and MitoTracker Red probe staining showed that the cell density in the drug group decreased in varying degrees,and a transition from branched to round morphology in the drug-treated groups was observed.The morphology of mitochondria tended to be round,and the ratio of the length of the longitudinal axis to transverse axis was(3.33±0.72)in the cell control group,(2.19±0.58)in the HM group,(2.45±0.44)in Mdivi-1 group,and(1.43±0.62)in HM+Mdivi-1 group,respectively.The results of JC-1 staining showed that compared with the cell control group,the mitochondrial mode potential of the HM group significantly decreased(P＜0.01).ROS significantly increased(P＜0.01)and ATP levels decreased(P＜0.01),and LDH enzyme activity increased(P＜0.01).Immunofluorescence staining and Western blotting results showed that compared with the cell control group,the expression levels of proapoptotic proteins Bax,cytochrome C,and caspase-3 in the HM group were significantly increased(all P＜0.01).Compared with the cell control group,the expression level of mitochondrial fission related protein Drp1 in HM group was significantly higher(P＜0.01).The expression level of mitochondrial fusion related protein Mfn2 significantly decreased(P＜0.01).After specific interference with Drp1 and synergistic intervention with HM,the survival rate of PC 12 cells in each interference group decreased compared to each drug intervention group.The expression of Drp1 and Mfn2 was downregulated,and the differences were statistically significant(P＜0.05 or P＜0.01).Conclusion HM can reduce the mitochoudrial membrane potential and ATP levels by accumulating ROS,there by activating the caspase-3 apoptosis pathway and promoting cell apoptosis.Mitochondrial fusion division may be involved in the damage of PC12 cells caused by HM,initiating apoptosis through the mitochondrial pathway.

Objective To explore the clinical significance of urine trace albumin in early diagnosis of diabetic nephropathy(DN). Methods A total of 35 patients with DN in DN group,32 patients with type 2 diabetes mellitus (DM)in DM group and 36 people accepted physical examination in control group.The urine trace albumin,blood urea nitrogen,serum creatinine were detected and compared.Results Urine trace albumin level in DN group was significant higher than those of DM group and control group,the differences were statistical significant(P <0.05),blood urea nitrogen and serum creatinine levels were slightly higher than those of DM group and control group,but there were no statistical significant differences (P >0.05).The urine trace albumin,blood urea ni-trogen and serum creatinine between DM group and control group had no statistical significant differences (P >0.05).Conclusion Urine trace albumin could be used as an early diagnostic index for early diagnosis of DN.

Objective To establish a method to prepare anti-sodium estrone sulfate monoclonal antibody ( ESS-Mab) . Methods Balb/c mice were immunized by ESS. Immune methods were screened. The blood serum potencies were measured by indirect ELISA and the best consistence of antigen and the first antibody were confirmed with method of titration. Cell fusion was carried by using PEG method and McAb hybridoma was screened with the indirect ELISA. Results The best immunization method of mice was subcutaneously multi-point injection in mouse back with the dose of 200/100 μg ESS antigen five times. The fusion rate was 90. 2%. Hybridoma positive rate of ELISA screening was 4. 4%. Finally two cell lines 2C8 and 8A7 with good specificity and sensitivity were obtained. Conclusion The best immunization way is selected and indirect ELISA is set up effectively and reliably for screening and presenting ESS McAb. the hybridoma technique is able to prepare monoclonal antibody of anti-ESS successfully.

Objective To explore the prognostic risk factors of low level malignant obstructive jaundice treated with transhapetic biliary drainage.Methods The clinical data of 142 patients with low level malignant obstructive jaundice received percutaneous transhapetic biliary drainage management from January 2010 to June 2013 were retrospectively analyzed.The study parameters included gender,age,tumor type,preoperative obstructive time,preoperative infection,drainage method,Child-Pugh grade,serum total bilirubin (TBIL),albumin (ALB),serum creatinine (SCr),the postoperative declining degree of bilirubin and postoperative antineoplastic therapy.The prognostic risk factors were evaluated.Results Single variable analysis showed that preoperative infection (P =0.006),Child-Pugh grade (P =0.004),SCr (P =0.043),the postoperative declining degree of TBIL (P =0.001) and postoperative antineoplastic therapy (P =0.015) were the related factors for survival time.The further Logistic regression analysis showed that preoperative infection (OR =3.729,95％ CI 1.332-6.363,P =0.040),Child-Pugh grade ≥ 10 scores (OR =0.513,95％ CI 0.375-1.276,P =0.018) and postoperative antineoplastic therapy (OR =0.668,95％ CI 0.210-2.026,P =0.038) were the related factors for survival time.Conclusion In treating of low level malignant obstructive jaundice with transhapetic biliary drainage,the preoperative infection,Child-Pugh grade and postoperative antineoplastic therapy may be the important related factors that affect the patient's survival time,to evaluate the prognosis of these patients has important reference meaning.

Objective To select the optimum conditions of Echinococcosis ELISA kit and kit preparation by using orthogonal experiments,then to test their detection performance.Methods Taking the absorbance of the samples as the indicator,orthogonal experiments of three factors four levels were taken to optimize the conditions of hydatid disease detection ELISA kit.Through L16 (43)orthogonal experiments,the concentration of the antigen(A),the dilution multiple of the sample (B) and the dilution multiple of the enzyme labeled antigen (C) were studied.The specimen were provided by The First Affiliated Hospital of Xinjiang Medical University specimen library collected between March 2011 and June 2013,including 36 sera confirmed alveolar Echinococcus by the gold standard,56 sera confirmed Echinococcus granulosus disease by the gold standard and 72 sera as the control group(including healthy people,cirrhosis,hepatitis,etc).Detecting sensitivity and specificity were compared using F test for statistical analysis.Results Orthogonal design showed the size proportion of three factors of echinococcosis ELISA kit:antibody dilution ＞ sample dilution ＞ antigen coating concentration.The optimal preparation conditions were A1 B2 C4,that was,the concentration of antigen was 1 mg/L,sample dilution of 1∶ 100,dilution of secondary antibody of 1∶160 000.Through the 3-factor and 4-level orthogonal design using F-test analysis,the fact of the concentration of the kit antigen(A),F =1.181,P =0.393; the fact of the dilution multiple of the sample (B),F =2.544,P =0.152,the fact of enzyme labeled antigen dilution (C) F =2.544,P =0.039.The sensitivity of the kit to Echinococcus granulosus disease and alveolar Echinococcus were 83％ (30/36),and 91％ (51/56),respectively; the specificity was 91％ (29/32) and 83％ (33/40); the misdiagnosis rate of 17％ (6/36) and 9％ (5/56) ; the misdiagnosis rate 9％ (3/32) and 18％ (7/40) ; positive likelihood ratio 8.86％ (83/9) and 5.20％ (91/18) ; negative likelihood ratio of 0.18％ (17/91) and 0.11％ (9/83).Conclusion Orthogonal design is a good method that can find out optimal conditions of preparation for Echinococcosis ELISA kit and improve the test performance.

Objective To observe the clinical curative effect of selective bronchial arterial embolization for severe hemoptysis.Methods Thirty-eight hospitalized patients with severe hemoptysis was enrolled in this study.Of which 31 patients were recurrent hemoptysis,27 patients were with bronchiectasis,4 patients were with bronchial lung cysts,4 patients had unknown causes,3 patients were with tuberculosis.All patients were diagnosed by chest imaging examination,fiber bronchoscopy,bronchial arteriography,and they were all treated by selective bronchial arterial embolization.Results The most times of embolization was 3,and bronchial arterial imaging were performed for vascular malformation.After having bronchial arterial embolization,35 patients were immediately released of bleeding.Postoperative 24 h,8 patients had hemoptysis again,of which 1 case was performed with conservative treatment and subsequently had pneumonectomy,5 patients had effective conservative treatment,2 cases were performed embolization again after failed conservative treatments.Hemoptysis completely disappeared within 1 week.The total effective rate was 97.4％(37/38).Patients were followed up for 1 year,of which 13 patients relapsed again,11 patients'annual and each time's quantity of hemoptysis reduced 50％.Two patients had pneumonectomy after a number of embolization.The total effective rate was 94.7％ (36/38).After treatment,3 patients had mild chest pain in short time,2 patients had shoulder pain,3 patients had chest tightness,and 3 patients had subcutaneous passive congestion.After being treated for short-term,these symptoms disappeared eventually.Conclusion Selective bronchial arterial embolization for the treatment of acute or recurrent severe hemoptysis is very effective,and can avoid the risk of surgery.It is effective for recurrent cases and worthy of clinical application.