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.

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.

In order to explore the role of acetylcholine in the pathogenesis of Parkinson's disease (PD), the changes in the concentration of acetylcholine (Ach) in the striatum, the apoptosis of substantia nigra cells, the ultrastructure and the changes of Nissl cells in rats during the morbidity of PD, and the corresponding behaviors in rats with PD were observed. Rat PD model was established by using the modified Thomas method. Eighty-one rats were randomly divided into normal control, sham operation and PD groups and their behavior features were observed at post-operative day (POD) 7, 14 and 21 as three subgroups (n=9 each). The concentration of Ach in the striatum was determined by using high-performance liquid chromatography. The apoptosis of substantia nigra cells was assayed by using TUNEL method. The ultrastructural changes in the substantia nigra were observed under the electron microscopy, and the survival of neurons in the substantia nigra area was examined by using Nissl staining. In PD group at POD 7 to 21, the damage in the substantia nigra area was gradually aggravated, the concentration of Ach, apoptosis rate and turns of rotation were gradually increased, and the number of Nissl cells was gradually reduced over the time as compared with the normal control and sham operation groups (all P<0.05). It was concluded that there exist dynamic changes in Ach concentration, ethology and apoptosis of the substantia nigra cells during the morbidity of PD, suggesting the contribution of apoptosis to the morbidity of PD, and critical role of Ach in the pathogenesis of PD.

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.

Objective:To explore the effect of family history of cancer on clinical features and prognostic factors in nasopharyngeal car-cinoma (NPC) patients. Methods:The clinical data of 89 NPC patients with a family history of cancer and 388 NPC patients without a family history of cancer were retrospectively reviewed. Univariate and multivariate survival analyses were performed to identify possi-ble prognostic factors. Results:The clinical characteristics of NPC patients with and without family history of cancer were compared. The gender, age, TNM stage, pathological type, and hemoglobin radiotherapy concentration before treatment did not significantly dif-fer between the two groups (P>0.05). NPC patients with a family history of cancer had better 3-year overall survival than those with-out family history of cancer (91.6%vs. 85.5%), but no statistically significant difference was observed (P=0.211). Both univariate and multivariate analyses showed that T, N, and TNM stages were the important prognosis factors affecting 3-year overall survival (OS), progression free survival (PFS), and distant metastasis-free survival (DMFS) of NPC (P<0.05). However, neither family history of cancer nor family history of NPC in 3-year OS, PFS, LRFS, and DMFS was significant in NPC patients (P>0.05). Conclusion:NPC patients with family history of cancer had better 3-year OS than those without family history of cancer, but no statistically significant observation was found. Large T stage or high lymph node stage contributed to poor survival of NPC. Family history of cancer had no significant in-fluence on the survival of NPC patients.

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 evaluate the changes of the waveform of the photopic negative response in flash-electroretinogram, visual acuity and central retinal thickness in the treatment of intravitreal injections of bevacizumub. Design Retrospective self-comparative case series. Partidpants 8 subjects (9 eyes) with exudative age-related macular degeneration and 3 subjects (3eyes) with proliferative diabetic retinopathy. Method Evaluation protocol included examinations of the Early Treatment Diabetic Retinopathy study visual acu-ity, visual field, intraocular pressure, fundus fluorescein angiography, optical coherence tomography and flash-electroretinogram. Intravit-real injections of bevacizumab, 1.25 mg (0.05ml), were given under an operating microscope and aseptic conditions. All the subjects were followed-up one month later. Main outcome Measure The amplitudes of PhNR, visual acuity and central retinal thickness. Re-sult At 1 months, the mean amplitudes of PhNR and mean visual acuity in all cases had no obvious change (n=12, P>0.05).The central retinal thickness reduced obviously (n=12, P<0.05), but it was neither significantly correlated with PhNR (r=0.294, P=0.145) nor with visual acuity(r=-0.358, P=0.073). Conclusion The single intravitreal injection of bevacizumab is showed promising in absorption of in-traretinal edema and subretinal fluid in patients with exudative age-related macular degeneration and proliferative diabetic retinopathy, but the changes of visual function (including PhNR) might need further investigation. (Ophthalmol CHN, 2009, 18: 243-246)

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.