The etiological agent of severe acute respiratory syndrome (SARS) was identified as a new coronavirus, termed SARS-CoV. Establishment of an efficient and sensitive diagnostic system of SARS-CoV genetic materials is crucial for SARS control. In this study, we quantified SARS-CoV mRNAs in both infected cell culture lysate and in supernatant by using Real-time quantitative revere transcription-PCR based on EvaGreen鈩?dye and Taqman-MGB probes. For extensive evaluation of sensitivities and specificities, 13 pairs of primers and 4 probes were designed based on different genes of SARS-CoV. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was selected as the internal control gene. Results showed that S-gene-specific PCR was the most sensitive for detection, but because of its sequence variability in the different viral strains, primers and a probe based on the N gene were suitable substitutions. Meanwhile, we found the mRNA concentrations in cell culture lysates were much higher than in cell supernatant and facilited more sensitive detection of the SARS-CoV.

A novel targeting drug carrier (FA-BO-PAMAM) based on the PAMAM G5 dendrimer modified with borneol (BO) and folic acid (FA) molecules on the periphery and doxorubicin (DOX) loaded in the interior was designed and prepared to achieve the purposes of enhancing the blood-brain barrier (BBB) transportation and improving the drug accumulation in the glioma cells. 1H NMR was used to confirm the synthesis of FA-BO-PAMAM; its morphology and mean size were analyzed by dynamic light scattering (DLS) and transmission electron microscope (TEM). Based on the HBMEC and C6 cells, cytotoxicity assay, transport across the BBB, cellular uptake and anti-tumor activity in vitro were investigated to evaluate the properties of nanocarriers in vitro. The results showed that the nanocarrier of FA-BO-PAMAM was successfully synthesized, which was spherical in morphology with the average size of (22.28 ± 0.42) nm, and zeta potential of (7.6 ± 0.89) mV. Cytotoxicity and transport across the BBB assay showed that BO-modified conjugates decreased the cytotoxicity of PAMAM against both HBMEC and C6 cells and exhibited higher BBB transportation ability than BO-unmodified conjugates; moreover, modification with FA increased the total uptake of DOX by C6 cells and enhanced the cytotoxicity of DOX-polymer against C6 cells. Therefore, FA-BO-PAMAM is a promising nanodrug delivery system in employing PAMAM as a drug carrier and treatment for brain glioma.

Objective To clarify the inhibitory effect of low molecular weight chitosan(LMCTS)on hepatic fibrosis induced by carbon tetrachloride (CCl4 )in the rats, and to investigate its effect on the expression of Toll-like receptor-4(TLR4 )and to lay the foundation for the development of the clinical candidate drug of liver fibrosis. Methods 72 male Wistar rats were randomly divided into blank control group, CCl4 group (model group), glycyrrhizinate (DG)group,50,100 and 150 mg·kg-1 LMCTS groups (low,middle and high doses of LMCTS groups).In addition to blank control group,the rats in the remaining groups were given 40% CCl4-vegetable oil (1.75 mL·kg-1 ),2 times per week for 8 weeks,by intraperitoneal injection to establish the model of rat hepatic fibrosis.And the rats in blank control group were injected with the same amount of 100% vegetable oil agent. From the ninth week, the rats in DG and LMCTS groups were given DG and LMCTS by intragastric administration, 1 time/week for 4 weeks. Then all rats were sacrificed, the activities of serum glutamic acid aminotransferase (ALT),aspartate aminotransferase (AST)and alkaline phosphatase (ALP)were detected with ELISA kit;the pathological changes in liver tissue were observed under light microscope, and the TLR4 expressions were detected by RT-PCR and Western blotting method. Results The serum ALT, AST and ALP activities in middle and high doses of LMCTS groups were lower than those in model group (P<0.05).The serum ALT activity in middle dose of LMCTS group was lower than that in low dose of LMCTS group (P<0.05),but the activities of AST and ALP had no statistically significant change(P>0.05).There were statistically significant differences in the serum ALT, AST and ALP activities between high dose of LMCTS group and middle dose of LMCTS group (P<0.05).There were obvious hepatocyte steatosis,inflammatory cell infiltration,collagen fiber hyperplasia and hepatic lobule damage in the rats in model group.However,all the changes in liver tissue of the rats in LMCTS group were significantly reduced, especially in high dose group. The results of RT-PCR and Western blotting method showed that the expression of TLR4 was declined in LMCTS groups compared with model group (P<0.05,P<0.01),especially in high dose of LMCTS group,and there were statistically significant differences between different doses of LMCTS groups (P<0.05).Conclusion High dose of LMCTS can decrease the serum transaminase activity of liver fibrosis rats and improve liver function,and this process may be related to declining the expression of TLR4 .

Objective To optimize spray drying process of Anshen bumian granules. Methods By using orthogonal design method, moisture content, powder rate and berberine hydrochloride content were indexes, and optimum technological conditions were determined. Results Proportion of microcrystalline cellulose, lactose and maltodextrin was 2??1??2. The extract was fed in at 6%. Optimum feeding in/out temperature for spray drying is 145 ℃ /85 ℃, liquid velocity was 30 mL??min-1 , and relative density of extract was 1. 15. Conclusion The process is reasonable and practical, has strong stability and good controllability, and it provides a basis for industrialized production.

Objective To research the fibrosis characteristics of Atrial Fibrillation(AF) patients under microscope and its effect to post-operative sinus conversion of Radiofrequency ablation treatment.Methods Based on Lefi Atrial Dimension (LAD).Left atrial appendage tissue samples of 60 AF patients who received bipolar radiofrequency ablation treatment during open heart surgery between July and December 2011,were divided into 4 groups:group a (LAD≤50 mm),group b (50 mm ＜ LAD≤60 mm),group c (60 mm＜ LAD ≤70 mm),group d (LAD ＞ 70 mm),15 patients in each group.The levels of atrial muscle fibrosis and cell size differences are compared for colored slides of samples.Research are done for the difference of sinus rhythm restoration rate among the 4 groups based on ECG records at immediate,leaving hospital,and 3,6,12 months postoperative are collected for all groups.Results Differences of CVF and atrial muscle cell size measured under microscope are statistically meaningful(P ＜ 0.001).Sinus rhythm restoration rate differences at 6 month(P =0.039) and 12 month (P =0.037) post-operative are statistically meaningful.Further,immediate,leaving hospital,and 3,6,12 months postoperative sinus rhythm restoration rates are 93.3％,93.3％,93.3％,100％,100％ for group a,80.0％,73.3％,80.0％,80.0％,80.0％ for group b,66.7％,66.7％,66.7％,73.3％,66.7％ for goup c,and 53.3％,53.3％,53.3％,60.0％,60.0％ for group d respectively.Conclusion For rheumatic heart valve diseases patients who at 6 and 12 month post-operative,the greater the LAD,the higher the atrial fibrosis level,the greater the cell size,the lower the sinus rhythm restoration rate is.

Objective To compare clinical outcomes of modified hand-assisted versus total laparoscopic splenectomy and esophagogastric devacularization for treatment of portal hypertension due to cirrhosis.Methods From Jan 2007 to Dec 2011,modified hand-assisted laparoscopic splenectomy plus esophagogastric devascularization (MHLSED) and total laparoscopic splenectomy and esophagogastric devascularization (LSED) were performed on 47 and 38 patients with portal hypertension due to cirrhosis,respectively.For the MHLESD group,we performed hand-assisted laparoscopic splenectomy first,then converted during operation to totally laparoscopic esophagogastric devascularization.The operating time,intra-operative blood loss,postoperative complications and postoperative hospitalization time were analyzed.Results MHLSED were performed on 47 patients successfully without any need to convert to open surgery,and LSED were performed on 36 patients with 2 patients having to convert to open surgery.The mean operative time [(154 ±32)min] and mean intra-operative blood loss [(115± 73)ml] in the HLSED group were significantly lower than the LSED group [(212±45)min and (172±57)ml,respectively].There was no mortality.There were no significant differences in the time period for gastrointestinal function to recover,postoperative hospital stay and overall complication rate between the two groups.Conclusions MHALSD is a relatively safe and efficacious treatment for portal hypertension due to cirrhosis.It combines the advantages of hand-assisted and totally laparoscopic operations.

ObjectiveTo investigate the effects of sleep deprivation on expressions of Mir-132,mir-134 in the different regions of rat brain.MethodsAll the male SD rats were divided into control group ( normal sleep group),sleep deprivation (SD).The modified multiple platform method (MMPM) was used to establish sleep deprivation model.Mir-132,mir-134 level was detected by real time PCR.ResultsMir-132 were significantly increased in SD groups in hippocampus compared with the control groups ( 51.87 ± 8.13 vs 67.25 ± 7.59 ) (P ＜0.01 ).Mir-134 were significantly decreased in SD groups compared with the control groups( 1.82 ±0.15 vs 1.45± 0.12 )(P ＜ 0.01 ).There were no statistically significant differences in cortex and thalamus (P ＞ 0.05 ).Cortex mir-132 level in SD group and control group was 1.57 ±0.10,1.48 ±0.11 respectively,and it was 1.37 ±0.09,1.36 ±0.11 in thalamus;Cortex mir-134 level in SD group and control group was 98.26 ± 5.17,100.80 ±4.15respectively,and it was 97.56 ± 6.28,91.01 ± 4.07 in thalamus.ConclusionThe upregulation of mir-132 and downregulation of mir-134 implies that two miRNAs did opposite actions in the processes of sleep deprivation.This findings indicate that hippocampus mir-132,mir-134 levels in the SD rat may reflect associated depressive patho-physiological processes.

ObjectiveTo evaluate the safety and efficacy of a modified hand-assited laparoscopic splenectomy (HALS) plus pericardial devascularization for the treatment of cirrhotic portal hypertension.MethodsFrom March 2009 to Dec 2011,modified hand-assited laparoscopic splenectomy plus pericardial devascularization was performed on 47 patients with portal hypertension and liver cirrhosis.We performed HALS first, thenconvertedtototallylarparocopicpericardialdevascularizationduring operation.ResultsAll patients received modified HALS plus pericardial devascularization without convertion to open surgery,the mean operative time was ( 154 ± 32) min,the mean intraoperative blood loss was ( 115 ±73) ml,and the mean postoperative hospitalization was (9.2 ± 1.6) days.The perioperative complications included plural effusion in 3 cases,ascites in 4 cases,pancreatic leakage in 1 case and wound dehiscence in 1case. Therewasnoperioperativemortality.ConclusionsModifiedHALSpluspericardial devascularization is a relatively safe and effective procedure in the treatment of portal hypertension due to liver cirrhosis,it has the advantage of hand-assisted and totally laparoscopic procedures.

Objective: To observe the effectiveness and safety of electroacupuncture (EA) plus Luohua Anshen oral liquid for patients with perimenopausal insomnia. Methods: A total of 66 participants who met the inclusion criteria were enrolled in the randomized controlled trial and allocated to a treatment group and a control group at a ratio of 1:1, with 33 cases in each group. Both groups were given Luohua Anshen oral liquid as a basic treatment. The treatment group was additionally given EA every other day, three times a week. Both groups were treated for four weeks and a four-week follow-up was conducted. The scores of Pittsburgh sleep quality index (PSQI), Kupperman index (KI) and traditional Chinese medicine sleep syndrome scale (TCMSSS) were recorded at pre- and post-treatment, and at the follow-up. Meanwhile, adverse effects were monitored and recorded. Results: After four-week treatment, the global scores of PSQI, KI and TCMSSS in both groups declined significantly (all P<0.05), and the decreases in the treatment group were more significant than those in the control group (allP<0.05). The global scores of PSQI, KI and TCMSSS in both groups at the follow-up visit were significantly different from the corresponding baseline (allP<0.05), while insignificantly different from those assessed at post-treatment (allP>0.05). The total effective rate was 93.9％ in the treatment group, significantly higher than 72.2％ in the control group (P<0.05). No significant adverse event was reported in this trial excepted one patient experienced slight dizziness in the first acupuncture treatment. Conclusion: EA plus Luohua Anshen oral liquid is safe for perimenopausal insomnia with satisfactory short- and long-term effectiveness, and it shows certain advantage compared with using Luohua Anshen oral liquid alone.

Objective To analyze the prognosis and risk factors for antineutrophil cytoplasmic antibody?associated vasculitis (AAV) patients on maintaining dialysis. Methods AAV patients on maintaining peritoneal dialysis (PD) or hemodialysis (HD) in First Affiliated Hospital Zhejiang University from June 2007 to June 2015 were included, and were followed up until death, kidney transplant, changed dialysis modalities or January 31, 2016. Patients were divided into PD group and HD group for comparison. Their survival rates and risk factors were analyzed by Kaplan?Meier analysis and COX regression model respectively. Results A total of 123 cases were chosen, with a median duration of dialysis for 854 (388, 1573) days, and with 88 cases (71.5%) on HD and 35 cases (28.5%) on PD. Fifty?two patients (42.3%) were more than 65 years old. At the median follow?up time of 36 months, 39 patients (31.7%) died. The main causes of death were cardiovascular events (30.8%) and infection (23.1%). COX regression analysis showed that patients older than 65 years old (HR=3.289, P=0.001), with cardiovascular disease (HR=3.241, P=0.003) and interstitial pneumonia (HR=2.173, P=0.048) at the dialysis onset were independent risk factors affecting survival. Conclusions Factors including age (older than 65 years), pre?dialysis cardiovascular disease and interstitial pneumonia were independent risk factors affecting survival of AAV patients on maintaining dialysis, then infections and cardiovascular events were the main causes of death.