Objective Respiratory syncytial virus (RSV) is the most common cause of lower respiratory infection in infants.It is very important to quantitative assay of RSV titer in the study on RSV pathogenesis,candidate vaccine and antiviral treatment.Therefore,we develop Real-time Quantitative PCR (Q-PCR) assay and enzyme immunospots (EIS) for titrating RSV and compare them with traditional 50% tissue culture infectious doses (TCID_(50)).Methods Q-PCR,based upon the RSV-L genes,and EIS were utilized to titrate samples from RSV culture supernatants and RSV infected mouse lungs.Then,the results were compared with TCID_(50).Results For the samples from RSV culture supernatants,the ratio of Q-PCR and EIS (plaque forming unit,pfu) was 10:1 and the ratio of EIS and TCID_(50) was 10:1 when TCID_(50) was converted as pfu.For the samples from RSV infected mouse lungs,the ratio of Q-PCR and EIS was 1000:1 and the ratio of EIS and TCID_(50) was 5:1.Conclusion We have successfully established Q-PCR and EIS to titrate RSV and compared them with TCID_(50).We concluded EIS is a cost-effective method to titrate RSV.

Objective To explore the impact of breast conservation surgery and modified radical mastectomy combined with intensity modulated radiation therapy respectively on the living quality and sexual life for patients with early-stage breast cancer.Methods From January 2011 to December 2016, 89 patients with breast cancer were admitted and divided into two groups according to different surgical methods.Patients in the conservation group (n=24) received breast conservation surgery combined with intensity modulated radiation therapy, while patients in the modified radical group (n=65) received breast radical surgery combined with intensity modulated radiation therapy.The quality of life and sexual satisfaction of patients were measured by questionnaire investigation.And the data were calculated by SPSS 16.0.Results Different surgical methods combined with radiotherapy affectted the score of living quality a lot.The average score was (87.500±7.940) points in the conservation group, while it was (65.350±8.490) points in the modified radical group, the difference was statistically significant (P<0.01).At the same time,the degree of self acceptance and sexual satisfaction in the conservation group after surgery was better than that in the modified radical group,and the difference was statistically significant (P<0.01).Conclusion Breast conservation surgery combined with intensity modulated radiation therapy for breast cancer patients could receive better quality of life and sexual satisfaction compared with modified radical mastectomy combined with intensity modulated radiation therapy.

To study the preventive effect of Grifola frondosa on nonalcoholic steatohepatitis (NASH). The rat model of NASH was established by feeding high-fat diets for 12 weeks and intervened with 0.5 g · kg(-1) · d(-1) and 1.0 g · kg(-1) · d(-1) of C. frondosa powder suspensions. The degrees of hepatocyte fatty degeneration and inflammation were observed under the optical microscope with routine HE staining. The NAFLD activity scores (NAS) were calculated. Serum ALT, AST and hepatic TG and CHOL were tested by the biochemical method. The hepatic MDA was examined by thiobarbituric acid method. The hepatic SOD was tested by the xanthine oxidase test. The hepatic GSH-PX activity was determined by the dithio-nitrobenzoic acid method. Hepatic TNF-α and IL-6 were detected by the enzyme-linked immunosorbent assay (ELISA). The NASH model group induced by high-fat diets showed higher hepatic NAS, ser- um ALT, AST, CHOL and hepatic TG, CHOL, MDA, TNF-α, IL-6 (P < 0.01 or P < 0.05) and lower serum TG and hepatic SOD, GSH-PX (P < 0.01, P < 0.05) than the normal control group. After being intervened with different doses of G. frondosa, the NASH group revealed significantly lower hepatic NAS, serum ALT and hepatic TG, CHOL, MDA, TNF-α and IL-6 (P < 0.05) and higher hepatic SOD, GSH-PX (P < 0.05) than the model group. G. frondosa may prevent the further development of NASH by improving the disorder of lipid metabolism in rats with NASH induced by high-fat diets, relieving the level of oxidative stress and reducing the generation of inflammatory cytokines.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Grifola ; chemistry ; Humans ; Interleukin-6 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

Study the serum level of HGF, Cys C and TGF-beta1 in type 2 diabetic nephropathy (DN), the infect of Pingshen decoction on those index. Selected 69 cases of 2 type DN and randomly divided into therapy group (36 cases) and control group (33 cases). The therapy group were treated with Pingshen decoction 1 dose/d, bid po. The control group were treated with NephritisShu tablet, 6 tablet, tid po. 8 weeks was a course. Before and after treatment, we examine the serum level of HGF, Cys C and TGF-beta1 by ELISA and immunonephelometry, and compare with 30 cases of healthy control group. The study demonstrates that before treatment, the serum level of HGF in both groups were significantly lower than healthy control group (P < 0.01), but Cys C, TGF-beta1 were significantly higher (P < 0.01). After treatment, the serum level of HGF of both groups were increased. The serum level of HGF of therapy group were significantly higher than of control group (P < 0.01), but the serum level of Cys C and TGF-beta1 were significantly lower than control group (P < 0.01). The serum level of HGF was correlated negatively with Cys C,TGF-beta1. In control group, the UAER, urine beta2-MG and quantity of 24-hour urine protein were significantly decreased after treatment (P < 0.01). The index of urine of therapy group were significantly lower than control group (P < 0.01). Results indicate that test of serum level of HGF and Cys C,TGF-beta1 of diabetic nephropathy have important clinical significance. Pingshen decoction can effectively intervene in the serum level of HGF and Cys C, TGF-beta1 and index of urine.
Aged ; Aged, 80 and over ; Case-Control Studies ; Cystatin C ; blood ; Diabetic Nephropathies ; blood ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hepatocyte Growth Factor ; blood ; Humans ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

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.

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.

The human epidermal growth factor receptor (EGFR) extracellular region (residues 1-621) consists of four subdomains, i.e. L1, S1, L2, and S2. The L2 domain (EGFR-L2) is composed of residues 311-479 and plays a major role in ligand-binding. Due to the high content of cysteine residues (42 cysteines) in the S1 and S2 domains, it is quite difficulty to get a correctly refolded product of the complete EGFR extracellular domain. In contrast, only 4 cysteine residues are present in EGFR-L2 domain. The aim of the present study is to prepare a soluble EGFR-L2 domain from the recombinant protein inclusion body overexpressed in Escherichia coli (E. coli). DNA fragment encoding EGFR-L2 containing a polyhistidine-tag at the carboxyl terminus was amplified by PCR from the cDNA of EGFR extracellular region, and was inserted into pET-3c to construct the prokaryotic expression vector. The target protein was highly expressed in E. coli BL21 (DE3) strain and was only present in the inclusion body as revealed by immunoblotting analysis. No soluble product could be refolded through dilution or stepwise dialysis strategies. However, on-column refolding of denatured EGFR-L2 bound to Ni2+ -NTA produced a soluble one. Furthermore,the soluble EGFR-L2 was simultaneously purified to high purity (>95%) through eluting from the same Ni2+ -NTA column with a linear imidazole gradient. The refolded EGFR-L2 had specific binding activity with the cognate ligand EGF, although its affinity was low. These results suggest that a polyhistidine-tag fused with a recombinant protein facilitate not only the purification but also the renaturation of the target product through on-column refolding. Besides, this refolding strategy may be suitable for the preparation of those recombinant proteins which are hard to refold through conventional approaches.
Escherichia coli ; genetics ; metabolism ; Humans ; Inclusion Bodies ; genetics ; metabolism ; Protein Folding ; Receptor, Epidermal Growth Factor ; biosynthesis ; genetics ; isolation & purification ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

The interaction of CD40 with its cognate ligand, CD40L (CD154), plays important roles in immune responses. Blockade of CD40-CD40L signal pathway can protect the progression of antibody- and cell-mediated autoimmune diseases, and reduce allograft rejection thus prolonging graft survival, even engendering long-lived antigen-specific tolerance. The present study aims to enhance the binding activity of CD40 by incorporating an isoleucine zipper (IZ) trimeric motif into CD40 ectodomain to promote the formation of soluble CD40 trimers, which would be useful for blocking CD40-CD40L interaction. A prokaryotic expression vector for soluble human CD40 ectodomain fused with an IZ motif and a hexa-histidine (His6) tag at its carboxyl terminus (sCD40IZ) was constructed by multiple round PCR using cloned CD40 cDNA as a template. The recombinant sCD40IZ protein was expressed highly in Escherichia coli (E. coli) with a molecular weight of 27kD, which is consistent with its theoretical value. It mainly existed in inclusion bodies. After refolding from inclusion bodies, soluble sCD40IZ protein was purified by gel filtration. Its molecular weight in solution was about 91kD when determined by gel filtration, suggesting that it most probably existed in the form of trimers. Moreover, this protein could bind to CD40L expressed on Jurkat T cells and its binding activity was significantly higher than that of soluble CD40 without an IZ motif. These results suggest that incorporation of an IZ motif at the carboxyl terminus of soluble CD40 can facilitate the formation of trimers and enhance its binding activity with CD40L. Thus, the trimeric CD40 protein may be used to block CD40-CD40L signal pathway, suggesting that it may have potential application in preventing autoimmune diseases and transplantation rejection.
CD40 Antigens ; biosynthesis ; genetics ; metabolism ; CD40 Ligand ; metabolism ; Escherichia coli ; genetics ; metabolism ; Humans ; Isoleucine ; biosynthesis ; genetics ; Leucine Zippers ; genetics ; Protein Multimerization ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; metabolism

The way of "extracting-salting-chromatography" was used to purify the phycoerythrin and phycocyanin from Porphyra yezoensis in process scale-up.First,by comprehensive comparison of efficiency,the Sephadex G-25 was selected from four resins (Sephadex G-25、G-100、S-300 and CL-6B) as the best choice used in crude extract desalting of phycobiliprotein.Then the preparation process of phycobiliprotein was scaled-up with raw material(Porphyra yezoensis) increased from 1g to 20g,and finally to 400g.The results indicated that the yields of purified phycoerythrin and phycocyanin (absorption spectra purity above 3.2) increased during according to process scale-up,with 0.323% phycoerythrin and 0.148% phycocyanin obtained from 400g frozen Porphyra yezoensis blades respectively.It is no doubt that the process involved in the experiment is a potential way for large scale preparation of phycobiliproteins of high purity.

To determine the role of vagi in heart rate variability, conscious rabbits were employed and electrocardiogram was recorded under conditions of bilateral vagi intact, unilateral vagotomy, and bilateral vagotomy. The variability of RR intervals (RRI) was analyzed using power spectrum and approximate entropy (ApEn). The results showed that the values of high frequency power (HF) component, low frequency power (LF) component and ApEn in animals with bilateral vagi intact were the highest, but the LF/HF ratio was the lowest; unilateral vagotomy decreased ApEn, right vagotomy increased LF/HF ratio but left vagotomy did not; the LF/HF ratio increased while ApEn decreased significantly in animals with bilateral vagotomy. It is suggested that the variability of RRI is mainly regulated by the vagi and the role of right vagi is more important than that of the left. When measuring heart rate variability, the results obtained with conventional method are consistent with those with nonlinear method.
Animals ; Entropy ; Heart Rate ; physiology ; Male ; Rabbits ; Vagus Nerve ; physiology