Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two-week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC).

To establish an HPLC mehod for the analysis of pharmacokinetics of salvianolic acid B in rats. METHODS: The biological samples were extracted with acetic ether. The chromatographic conditions were as follows: Hypersil ODS column (200 mm×4.6 mm, 5μm) was used. The mobile phase was acetonitrile-water(with Ammoniom Acetate 0.25 mol/L) was set at 328 nm. RESULTS: Salvianolic acid B was injected intravenously at doses of 1.6, 3.2, 6.4 mg/kg. The terminal elimination half-life(t1/2) of α phase and β phase was (3.1±0.1) min and (31.5±3.2) min. The extents of excrement,urine and biliary excretion of salvianolic acid B were 1.43%±0.90%, 0.77%±1.01% and 8.82%±4.11%. The tissue concentration of salvianolic acid B was as followed in order: Cheart>Cliver>Clung>Cintestine>Ckidney>Cspleen>Cstomach. The plasma protein binding rate of salvianolic acid B in human plasma and in rat was similar(89.2%±1.8%,92.5%±1.5%). CONCLUSION: The method is accurate, stable and reliable, and can be used for the investigation of salvianolic acid B in pharmacokinetics research. Salvianolic acid B eliminates fast and it shows a high plasma protein binding rate, the mainly excretion way of salvianolic acid B is from biliary.

Aim To study the effect of proteasome inhibitor MG132 on expression of Caspase-3 and UPP associated genes in the apoptosis of human hepatocelluar cancer cells.Methods HepG2 cells were treated with MG132 (2,5,10 ?mol?L -1) for 24 h. The apoptotic cells were determined with flow cytometric analysis. The levels of E1, E2, E3 and Caspase-3 mRNA expression were detected with RT-PCR. Caspase-3 protein expression was detected with immunohistochemistry. Results The results showed that the increase of the degree of HepG2 cell apoptosis was concentrationly dependent. RT-PCR showed that E1, E2 and E3 gene expressions were decreased in the treatment group. MG132 down-regulated the gene expression of E1, E2, E3 and up-regulated the gene/protein expression of Caspase-3. Conclusion Proteasome inhibitor MG132 may induce HepG2 cells apoptosis by inhibitting UPP activity,up-regulating the gene/protein expression of Caspase-3.

AIM: To discuss the clinical efficacy of fresh amniotic membrane ( AM ) during the microscopic adjustable suture surgery in children's intercommunity strabismus, in order to guide clinical treatment.
METHODS: With the clinical randomized control study (RCT), 60 (112 eyes) cases of patients in childhood who received microscopic strabismus surgery in our hospital were divided them into two different groups from Jan. 2010 to Oct. 2015. According to the application of AM on the basis of ophthalmology outpatient number, 30 cases (58 eyes) in group A were treated with rectus muscle recession surgery combined adjustable suture combined with AM. The other 30 cases (54 eyes) in group B were treated with rectus muscle recession surgery combined adjustable suture only. All patients in two groups were followed-up over 6mo after the strabismus surgery.
RESULTS:Twenty-seven cases ( 48 eyes ) of all the strabismus patients must be adjusted after strabismus surgery, and the eye position adjustment rate was 42.9%. At 1mo after surgery, eye position of 18 cases (29 eyes) can be adjusted in all patients, and 44. 8% (16 cases, 26 eyes ) in group A with the average of adjustment lengths was 2. 56±0. 64mm, and 5. 6% ( 2 cases, 3 eyes ) in group B, with the average of adjustment lengths was 0. 52±0. 28mm, the differences of the adjustment rate and the average of adjustment amount were both high statistically significant (χ2 =22.477, P<0. 01; t=16. 502, P<0. 01 ) between the two groups. Except of 3 cases who couldn't cooperate with eye position adjustment, they all received eye position adjustment in different degrees in one month after strabismus surgery,and after eye position adjustment, 27 cases (53 eyes) in group A got normal eye position, and the correction rate of eye position was 91. 4%, and 16 cases (28 eyes) in group B got normal eye position after eye position adjustment, the correction rate was 51. 9%, the differences of the correction rate were statistically significant (χ2=21. 827, P<0. 01) between the two groups.
CONCLUSION: The application of fresh AM in the microscopic adjustable suture strabismus surgery is exactly effective in treatment of children's intercommunity strabismus. It can significantly extend the adjustment time and increase the adjustment amount, and it also can statistically improve the controllability and achievement ratio for children's strabismus surgery.

This review introduced the anti-tumor effects of non-steroid anti-inflammatory drugs (NSAIDs) and summarized their possible molecular mechanisms according to recent abroad literatures and our research results. Some evidence showed that the anti-tumor mechanisms of NSAIDs were different in various tumors.NSAIDs decreased the biosynthesis of PGE_2 and regulated the expressions of downstream correlated genes and proteins through restraining abnormal expression of COX-2 in certain neoplasms,which resulted in the inhibition of tumor angiogenesis and proliferation as well as induced apoptosis. But in other cancer cells, NSAIDs, as activators of peroxisome proliferator-activated receptor ? (PPAR?), induced COX-2 expression, promoted the biosynthesis of cyclopentenone prostaglandins (cyPGs). cyPGs further induced tumor cell apoptosis with PPAR? dependently or PPAR? independently. Since their special mechanisms of anti-proliferation and pro-apoptosis, NSAIDs revealed significant synergistic effects with other anti-tumor treatments.

Objective To explore the clinical efficacy and safety of transcatheter absolute ethanol injection treatment on varicose veins of lower extremity.Methods twenty-there patients with 25 varicose veins of lower extremity were treated by puncture of great saphenous vein above 1—2 cm of complicated inner ankle,perforating catheter to the point below the 3—4 cm of the conjunction of great saphenous vein and Femoral vein and pressing the conjunction of these two veins.Under the monitor of DSA,inject the absolute ethenal slowly while retrieve the catheter little by little(one limb with varicose veins injected total volume 15—20 ml),in the mean time,using contrast agent to monitor the level of embolism until the formation of total embolism in the all great saphenous veins.Results All the cases were retrospectively followed up with CDFI examination after 3—12 months of the surgery,No blood flow were seen in the 25 embolismic great saphenous vein.Clinical symptom were alleviated obviously after 2—3 weeks of treatment;varicose veins were collapse after 3 to 7 days.Two eases of leg ulceration were healed after 4 to 6 weeks of operation.20 limbs were found mild swelling in the 2 day after the surgery.However,all the cases were disappeared after 1 to 2 weeks;4 treated limbs developed delayed paresthesia in the 3 day after the surgery,and recovered totally in the 2 weeks.No complications of deep vein thrombosis,lung thrombosis etc al,were found after operation.Conclusions Using transcatheter injection of absolute ethanol to treat varicose veins of lower extremity has the advantage of less invasion,more safety and low appearance of complications.The short term efficacy is solid while the long term effect needs further evaluation.

OBJECTIVE: To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema. METHODS: The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4. RESULTS: Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased. CONCLUSION The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.
Animals ; Aquaporin 4/metabolism* ; Astrocytes ; Blood-Brain Barrier/metabolism* ; Brain ; Brain Edema/metabolism* ; Brain Injuries/metabolism* ; Cell Membrane Permeability/genetics* ; Disease Models, Animal ; Permeability ; Rats ; Water

AIMTo study the effects of various liposomes formulations and preparation methods on the stability of acyclovir palmitate (ACV-C16) liposomes on storage at 4 degrees C and 25 degrees C over a 6 months period.

METHODSThe mean particle size, Zeta potential, pH and leaking ratio of ACV-C16 liposomes were the parameters chosen to indicate the stability of liposomes. All of the parameters were compared among various lipid compositions [egg lecithin/cholesterol/hosphatidylserine (PC/CH/PS), egg lecithin/cholesterol/stearylamine (PC/CH/SA), egg lecithin/cholesterol/cholesteryl sulphate (PC/CH/CS), bovine brain ceramides/cholesterol/palmitic acid/cholesteryl sulphate (CM/CH/PA/CS)], different preparation methods (film dispersing, reverse phase evaporation, dehydration/rehydration), charges (positive, negative), as well as among multilamellar vesicles liposomes (MLV), large unilamellar vesicles liposomes (LUV) and dehydration/rehydration vesicles liposomes (DRV).

RESULTSAn analysis of various parameters led to the conclusion that the stability of liposomes followed the order of PC/CH/CS > CM/CH/PA/CS > PC/CH/PS > PC/CH/SA at the same storage conditions; the positively charged system showed the most unstable delivery system of liposomes as compared to the other three systems. As far as stability was concerned, LUV liposomes proved to be superior to MLV liposomes and DRV liposomes, and the modified reverse phase evaporation method of Szoka provided the best preparation method. The stability in systems was enhanced when systems were stored at 4 degrees C as compared to storage at 25 degrees C.

CONCLUSIONThe stability of liposomes was significantly interrelated with lipid composition of various liposomes, preparation method and different storage conditions.

Acyclovir ; administration & dosage ; Chemistry, Pharmaceutical ; Drug Carriers ; Drug Delivery Systems ; Drug Stability ; Liposomes ; chemistry ; Palmitates ; chemistry ; Particle Size ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate effectiveness of scopolamine on the acute severe chlorphenamidine poisoning patients.

METHODS72 cases of acute severe chlorphenamidine poisoning patients were divided into I and II groups by the principle of a 1:1 sampling according to the order of admission. The I group (36 cases) were treated with traditional multimodality therapy, including gastrolavage, catharsis, using reductant-oxidant (methylthioninium chloride and vitamin C), and symptomatic treatment. The II group were treated with traditional multimodality therapy and scopolamine at the same times. Blood methemoglobin were measured at 0, third, seventh, twelfth, twenty-fourth hour, serum troponin I (CTnI) and creatine kinase isoenzyme (CK-MB) levels at third, seventh, twenty-fourth, forty-eighth hour, hepatic and renal functions at third, twenty-fourth, forty-eighth hour, and electrocardiogram (ECG) were evaluated every 4 hours in 3 days after hospitalization on all patients. The two groups of patients were compared the efficacy and change detection of targets.

RESULTS31 patients (86.11%) recovered and 5 patients (13.89%) died in I group. All 36 cases recovered in II group. The recovery rate of II group was distinctively higher than that in I group (P < 0.05) and the difference was statistically significant (P < 0.05). The average recovery time and the length of hospital stay in II group were sharply shorter than those in I group (P < 0.01) and the difference was statistically significant (P < 0.05). Serum CTnI levels between seventh hour and forty-eighth hour, serum CK-MB levels between third hour and forty-eighth hour and methemoglobin concentration at third, seventh, twelfth, twenty-fourth hour were apparently lower in II group, and the difference was statistically significant (P < 0.05). The abnormal rates of hepatic and renal functions in II group were distinctively lower than those in I group and the difference was statistically significant (P < 0.05). The abnormal rates of ECG in the second and third day in II group were respectively 38.89% and 11.11%, and were lower than those in I group (64.71%, 38.71%). The difference was statistically significant (P < 0.05).

CONCLUSIONScopolamine has the excellent treatment effect on acute severe chlorphenamidine poisoning patients and protec their hearts, livers, and kidneys. It complements the deficiency of reductant-oxidants, and combination of the two drugs can form the synergy effect.

Adult ; Chlorphenamidine ; poisoning ; Female ; Humans ; Male ; Scopolamine Hydrobromide ; therapeutic use ; Treatment Outcome

Animals ; Captopril ; pharmacology ; Heart Failure ; metabolism ; Male ; Myocytes, Cardiac ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism