Animals ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; pharmacology ; Humans ; Research Design

Congenital Abnormalities ; diagnosis ; therapy ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans

Objective To observe the transport of hepatitis B virus (HBV)-infected peripheral blood mononuclear cells (PBMC)through placental barrier set up by choriocarcinoma trophoblast cells (Bewo cells),and to explore the biological role of PBMC as a carrier for HBV transport.Methods Bewo cells and PBMC were cultured and their proliferation and activity were detected by cell counting kit (CCK)-8.One hundred μL serum containing 5 ×10 6 copy/mL HBV DNA was used to infect PBMC,and cells infected with HBV were labeled by fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE).A co-culture model of Bewo cells and HBV-infected PBMC was set up by transwell chamber. The migration of HBV-infected PBMC was detected by flow cytometry.Realtime fluorescence quantitative polymerase chain reaction method was used to detect HBV DNA contents of PBMC under transwell chamber.Results PBMC and Bewo cells proliferated at around 24 h and entered into growth stagnation at around 120 h.The contents of PBMC labeled by green fluorescent at 0,12,24 and 48 h during co-culture under chamber were (0.445 ±0.021)%,(21 .180 ± 4.653 )%,(34.830 ± 7.156 )% and (64.185 ± 3.161)%,respectively.The amount of PBMC marked green fluorescence increased over prolonged incubation time (F =68.983,P =0.001 ).PBMC HBV DNA contents at 24 and 48 h of co-culture under chamber were (1.925±0.431)×103 copy/mL and (2.565 ±0.361)×103 copy/mL,respectively,indicating that PBMC under chamber were infected with HBV.Conclusions PBMC may be a target for HBV infection in extrahepatic tissues.Placental trophoblastic barrier built by transwell chambers may provide new ideas to investigate HBV transmission across the placenta in vitro .

OBJECTIVE:To provide suggestions for the government decision of payment price of medicare drugs. METHODS:Investigation was conducted for the beginning of payment price policy of medicare drugs,effect of the implementation of policy in other countries and the encountered problems. The payment pricing mechanism was explored based on the combination of new medi-cal reform with theoretical analysis and empirical research. RESULTS:The reform of payment pricing mechanism of medicare drugs will have effect on current drug price,especially for R&D oriented pharmaceutical industries. The policy can decrease drug expendi-ture in medicare expenditure,and it is not sure whether it can decrease total medicare expenditure. CONCLUSIONS:It is suggest-ed to notice the related measures and government should keep balance between drug availability and encouraging R&D innovation.

Objective To observe the anti‐metastatic effect and mechanism of baicalin on the growth of HeLa cells was measured by MTT assay ,and cell migration baicalin on human cervical cancer HeLa cells .Methods The effects of baicalin on the proliferation and invasion of HeLa cells were analyzed by MTT method and Transwell assay .Moreover ,Real‐time PCR was used for investigating the expressions of MMP‐2 and MMP‐9 at the RNA level .Western blot was used for investigating the expressions of MMP‐2 ,MMP‐9 ,P38 and p‐P38 at the protein level .Results Baicalin could significantly inhibit the proliferation of HeLa cells in the dose‐dependent manner at the concentration above 60 μg/mL . Anti‐metastatic signaling induced by baicalin was characterized by down‐regulating the RNA and protein expressions of MMP‐2 and MMP‐9 ,and down‐regulating the phosphorylation level of P38 . Pre‐treatment of P38 signal pathway inhibitor could enhance the inhibitory effect of baicalin on the expressions of MMP‐2 and MMP‐9 .Conclusion These results indicate that baicalin‐induced anti‐metastatic effect involves the inhibition of MMP‐2 and MMP‐9 in HeLa cells through P38 signal pathway .

Objective To evaluate the effects of hepatitis B virus on liver function,liver fibrosis,and liver pathological staging at different immune stages.Methods We made a retrospective analysis of 657 patients with chronic hepatitis B diagnosed in the First Hospital of Lanzhou University.Their liver function parameters,liver fibrosis parameters,and hepatitis B virus load were measured by automatic biochemical analyzer,automatic gammaradiation immunity analyzer,and quantitative PCR analyzer,respectively.Effects of hepatitis B virus on liver function,liver fibrosis in different immune stages were analyzed by variance analysis.Effects of hepatitis B virus on liver pathological staging at different immune stages were analyzed by linear trend chi square test analysis.Results In ALT normal chronic hepatitis B patients group,viral load had mild effects on liver function and liver fibrosis parameters.However,in ALT abnormal chronic hepatitis B patients group,viral load had a significant effect on liver function and liver fibrosis parameters,and the effect was most obvious in ALT＞double upper limit of normal group.The specific manifestation was that with viral load increasing,liver function parameters including ALT,AST,TBiL,DBiL,and IBiL increased,while TP and ALB decreased.Liver fibrosis parameters HA,LN,PcⅢ,and CIV all increased (P＜0.05).In ALT normal chronic hepatitis B patients group,viral load had no relationship with liver pathological staging.However,in ALT abnormal chronic hepatitis B patients group,especially ALT≥double upper limit of normal group,viral load was significantly related to liver pathological staging.Conclusion The effects of hepatitis B virus on patients' liver function at different immune stages were different,thus providing evidence-based medicine support for clinical antiviral treatment.

Objective To evaluate the efficacy of dual stent placements for the treatment of occlusion/stenosis of subclavian artery associated with stenosis of adjacent vertebral artery initial site. Methods The clinical data of 9 patients with occlusion/stenosis of subclavian artery associated with stenosis of adjacent vertebral artery initial site were retrospectively analyzed. Stent implantations in the affected subclavian artery and vertebral artery were separately performed; the patients were followed up for 3-12 months after the treatment. The therapeutic efficacy was evaluated with the clinical symptoms and Doppler ultrasonic examination. Results Successful implantation of two stents was accomplished in all 9 patients, with the technical success rate of 100%. After the treatment, the blood flow in both the subclavian and vertebral arteries was unobstructed. Following-up examination showed that the subjective symptoms were obviously improved in all 9 patients, and no serious procedure-related complications occurred. Doppler ultrasound examination showed that no in-stent restenosis or stent displacement was observed. Conclusion For the treatment of occlusion/stenosis of subclavian artery associated with stenosis of adjacent vertebral artery initial site, dual stent placement technique is a safe and feasible means with satisfactory effect in improving vertebro-basilar arterial insufficiency.

Objective To analyze the clinical manifestations,diagnosis and treatment of idiopathic left ventricular tachycardia(ILVT)in infants and young children.Methods The clinical data,including manifestations,lab results and treatments of 37 cases of ILVT in infants and young children were analyzed retrospectively.Results ILVT were seen more in young children older than 1 year old and male infants.A total of 37 cases with ILVT showed diverse clinical manifestations,5 children had cardiac insufficiency with oliguria and edema,2 children had syncope.There were 8 children without any symptoms,and 22 children had some symptoms,which mainly included agitation and crying,weakness and sweaty,vomitting and pale com-plexion.Holter showed incessant ILVT in 20 cases(54.1%),and paroxysmal in 17 cases(45.9%).Of the 37 cases,9 cases presented tachycardia-induced cardiomyopathy(TIC),8 of them were secondary to the in-cessant ILVT.Children with TIC compared with those without TIC,the ventricular rate were faster[(206.2 ± 38.7)beats/min vs.(171.8 ±38.7)beats/min,P<0.05],the duration were longer[(22.6 ± 15.3)days vs. (9.8 ±6.0)days,P<0.05].A total of 34 cases were given by antiarrhythmic drugs,verapamil was higher effective in reversion to sinus rhythm.Radiofrequency ablations were conducted in 8 cases,resulting in ven-tricular tachycardia elimination.All of children were fine during following up periods.After successful control of tachycardia,left ventricular end-diastolic diameter and left ventricular ejection fraction in patients with TIC gradually recovered[(35.4 ± 7.2 )mm vs.(28.9 ± 5.6 )mm,P <0.05;(46.7 ± 4.0 )% vs.(70.1 ± 1 .5 )%,P<0.0 1 ] .Conclusion ILVT in infants and young children are rare,and present different manifes-tions.An incessant ILVT with faster ventricular rate and longer duration are more frequently complicated by TIC.Radiofrequency ablation is the effective way of radical cure of the disease.The diagnosis and treatment for the tachycardia are particularly important in early time,and the prognosis are good.

Objective To observe the protective effects of roscovitine on the podocyte injury induced by endoplasmic reticulum stress (ERS) caused by tunicamycin. Methods The differentiated podocytes cultured at 37℃were randomly di-vided into:(1) Control group, DMSO group and tunicamycin group (TM, 1.0μmol/L). The treatment was given for 3, 6 and 12 hours in three groups. (2) For control group, tunicamycin group, tunicamycin+roscovitine group (20, 40μmol/L, TM+ROS), the treatment was given for 12 hours. The podocyte apoptosis was detected by flow cytometry and TUNEL method. The ex-pressions of Cdk5, GRP78, Caspase-12 and CHOP were detected by Western blot assay. Results (1) Compared with con-trol group and DMSO group, the podocyte apoptosis was increased significantly in a time dependent manner after tunicamy-cin treatment in TM group;the protein expressions of Cdk5, GRP78, Caspase-12 and CHOP were also up-regulated signifi-cantly in TM group (P<0.05). (2) Flow cytometry and TUNEL analysis showed that tunicamycin induced apoptosis in podo-cytes, which was significantly inhibited by roscovitine in a concentration dependent manner in TM+ROS group as compared to that of TM group (P<0.05). The protein expressions of GRP78, Caspase-12 and CHOP were also significantly decreased in a concentration dependent manner in TM+ROS group compared to those of TM group (P<0.05). Conclusion Roscovi-tine, the inhibitor of Cdk5, can reduce the podocyte apoptosis induced by tunicamycin. The protective effects of roscovitine on podocytes can be a novel approach of treating diabetic nephropathy.

Objective:To investigate roles of autophagy in ameliorating sepsis-induced acute lung injury by allicinin in mice.Methods:A total of 152 male Balb/c mice (8-week old) were randomly divided into a sham group,a septic model group,an allicin treatment group,and an autophagy inhibition group.Septic mouse model was established by cecal ligation and puncture (CLP).Mice in the allicin treatment group were given allicin (30 mg/kg,intra-peritoneal injection) at 6 and 12 h,while those in the autophagy inhibition group were given autophagy inhibitor 3-MA (15 mg/kg,intra-peritoneal injection) at half an hour after allicin administration.Mice in the model and sham group were administered with the same amount of saline.Twenty mice in each group were randomly chosen to observe the 7 d survival rate.The other 12 mice were killed at 24 h,and the bronchoalveolar lavage fluid (BALF) (n=6) and lung tissues (n=6) were collected.ELISA was used to detect the tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the BALF.Hematoxylin-eosin staining was preformed to show the morphological changes in the lung tissues.Malondialdehyde (MDA) content and the activity of superoxide dismutase (SOD) in the lung tissues were examined.The expression of LC3B and Beclin-1 was determined by immunohistochemical analysis.Results:Compared with the sham group,the 7 d survival rate and lung SOD activity were decreased in the CLP group (P<0.05);the lung morphological damage score,the levels of TNF-α and IL-6 in the BALF,MDA content in the lung,and expression of LC3B and Beclin-1 were increased greatly in the CLP group (P<0.05).Compared with the CLP group,the 7 d survival rate,lung SOD activity and the expressions of LC3B and Beclin-1 were increased significantly in the allicin treatment group (P<0.05);the lung morphological damage scores,the levels of TNF-α and IL-6 in the BALF and MDA content in the lung were decreased obviously in the allicin treatment group (P<0.05).Compared with the allicin treatment group,the 7 d survival rate,lung SOD activity,and the expressions of LC3B and Beclin-1 were decreased in the 3-MA group (P<0.05);the lung morphological damage scores,the levels of TNF-α and IL-6 in the BALF,and MDA content in the lung were increased significantly in the 3-MA group (P<0.05).Conclusion:Allicin may ameliorate sepsis-induced acute lung injury in mice by enhancing the level of autophagy.