Hyperplasia of smooth muscle cells is a major aspect of airway remodeling in asthma .In recent years.It is found that the epigenetic regulation plays an important role on the proliferation of airway smooth muscle cells and the secretion of inflammatory cytokines , including DNA methyltransferase inhibitors to inhibit the phenotype trans-formation;histone acetylation related with hypertrophy .In addition , the microRNA is potentially related with the regulation of a variety of physiological functions of airway smooth muscle cells of asthma model , including inhibition of proliferation and release of inflammatory factors .Hope that epigenetics can become a new target for the treatment of asthma .

In recent years, with the rapid development of antibody drugs, the antibody-based therapies have gradually expanded from the cancer and autoimmune diseases to metabolic and infectious diseases and so on. However, the development of antibody-based anti-infective drugs is much slower as there are only two kinds of drugs in the market. This is due to the complex infective mechanism of viruses, bacteria and other pathogens, and the monovalent character of monoclonal antibodies that greatly limit the anti-infection effect of antibody drugs. The development and application of novel technologies, such as recombinant polyclonal antibody technology, will greatly accelerate the development of antibody-based anti-infection drugs. This article will introduce the application and trends in the development of antibody-based drugs in the field of anti-infection therapy.
Anti-Infective Agents ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Drug Design ; Humans

Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Purpura, Thrombotic Thrombocytopenic ; etiology ; Transplantation, Autologous

Objective:To observe the clinical effects of acupuncture plus external medicine application for cervical radiculopathy.Methods:A total of 98 patients with cervical radiculopathy were randomly divided into an observation group and a control group based on the random digital table.The observation group (50 cases) was treated by acupuncture plus external medicine application and the control group (48 cases) was treated by acupuncture alone.The patients received the treatment every day.Ten sessions made a course.Cervical spondylosis symptom scale was used to assess the cases before and after the treatment.The clinical efficacy was compared between the two groups after the treatment.Results:The total effective rate was 98.1％ in the observation group versus 87.6％ in the control group with a statistically significant difference between the two groups (P＜0.05).The cure rate was 52.0％ in the observation group versus 18.8％ in the control group with a significant difference between the two groups (P＜0.05).There was no significant difference in the score of cervical spondylosis symptom scale between the two groups before the treatment (P＞0.05).It increased with a significant difference in both groups after the treatment (both P＜0.05),while a significant difference was found in the results between the two groups after the treatment (P＜0.05).Conclusion:Acupuncture plus external medicine application is more effective than acupuncture alone for treating cervical radiculopathy.The method also effectively improves the clinical symptoms.

Objective To construct and identify the efficacy of a lentiviral vector harboring RNAi sequence targe-ting NSBP1 gene. Methods Three siRNA targeting the NSBP1 mRNA were designed, the pGCSIL-GFP-NSBP1 lentivirus vectors were constructed and confirmed by DNA sequencing. A total of 293T cells were co-transfected with pGCSIL-GFP-NSBP1, pHelper1.0 and pHelper2.0 for the virus stocks produced, the titer of the virus was test-ed. After lentivirus transfecting into DU145 ceils, Western-blot and MTT methods were used to determine the ex-pression and biological activity of NSBP1 gene, the cells were transplanted into nude mice, then inhibitive effect was observed. Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the hu-man NSBP1 gene was successfully inserted into the lentiviral vector. The titer of the recombinant]entiviral vector was 2 × 10~8TU/mL. NSBP1 protein expression level in transfected cells was significantly decreased and growth rate of cells transfected with lentivirus was decreased by MTT assay, the downregulation of NSBP1 reduced growth rate of transplantated tumor, whereas tumorgenicity was not influenced. Conclusion The construction of the]entiviral vector of NSBP1 has been successfully prepared and NSBP1 plays an important regulatory role in androgen-inde-pendent prostate cancer cell proliferation.

Objective To assess the diagnostic value of CT-gnided pereutaneous fine-needle aspiration biopsy (FANB) for the space-occupying lesions of the pancreas. Methods The authors retrospectively reviewed the medical records of 68 patients with pancreatic lesions who underwent CT-guided percutaneous fine-needle aspiration biopsy. The lesions were located at the pancreatic head (n = 49), body (n = 12) or tail (n = 7). The maximum diameter of the 68 lesions ranged from 2 cm to 7 cm. The lesion's diameter was < 3 cm in 10 cases and was ≥ 3 cm in 58 cases. A 20-gauge fine-needle was used to puncture and to make the aspiration in all patients. Both plain and contrast-enhanced abdominal CT scanning were performed before the biopsy procedure. The technical key points of aspiration biopsy included selection of the appropriate cross-section, the entry of the needle, and multiple insertions in fan-like fashion with fast manipulation of puncture and aspiration. All biopsy specimens were sent for pathologic evaluation. Results All the lesions were safely and successfully punctured through the anterior approach without major complications. The technical success of needle puncturing was 100%. Of 68 patients, the pathological results obtained from the biopsy included malignant tumor in 46, benign lesion in 17 and no abnormal finding in 5, of which follow-up observation found no abnormality in 2 and malignant process was eventually confirmed in remaining three. The overall accuracy, specificity and sensitivity of the biopsy were 96%, 95% and 100%, respectively. The accuracy of CT-guided biopsy for malignant and benign lesions was 94% and 100% respectively (P > 0.05). The diagnostic accuracy for lesions larger than 3.0 cm (97%) or located at the pancreatic tail (100%) was a little higher than that for lesions smaller than 3.0 cm (90%) or located at the pancreatic head (96%) or body (92%), although the difference was not statistically significant (P > 0.05). No serious complications occurred in our series. Conclusion CT-guided percutaneous fine-needle aspiration biopsy is a safe and effective technique for the diagnosis and differentiation of the space-occupying lesions of the pancreas.

Objective To investigate the character and location of a novel gene R049 and its expressed protein in uropathogenic Escherichia coli(UPEC) strain 132 isolated in China. MethodsThe chromosome library of UPEC132 was constructed by a shotgun strategy and the sequence analysis was carried out by a high-throughput pyrophosphate sequencing. Sequence reads were assembled with the Newbler program.The characters of R049-associated specific fragment were analyzed using the bioinformatics methods. Outer and inner membrane proteins of UPEC132 were extracted and then detected by SDS-PAGE and Western blot analysis together with the whole-cell lysates. ResultsThe 169 022 bp contig containing gene R049 was obtained and its sequence was very similar to the chromosome associated sequence of UPEC strain 536. It showed that a 20 773 bp fragment including R049 replaced the pathogenicity island PAI Ⅲ536 of UPEC536 in above 169 022 bp contig. The fragment had a lower GC content (46.97％) and 16 bp direct repeats in two ends. Significantly it also was adjacented to thrW tRNA, insertion element and genes coding integrase. Thus the 20 773 bp fragment was named R049 genome island(R049-GI). There were 25 ORFs in R049-GI, and gene R049 was located in the thirteenth ORF. The results of SDS-PAGE and Western blot revealed gene R049 encoded an outer membrane protein in the size of 47.0× 103. ConclusionGene R049, encoding an outer membrane protein, was a component part of the genome island in UPEC 132 chromosome acquired by horizontal gene transfer.

Objective To investigate the effect of angiotenisn ⅡI (Ang Ⅱ) on RIN-m β-cell,and to explore the mechanism of β-cell function impairment caused by Ang Ⅱ.Methods RIN-m cells were cultured with various concentrations of AngⅡ (0.1,1,10,100 nmol/L).After incubation for 24 hours,the basal(3.3 mmol/L) and glucose-stimulated(16.7 mmol/L) insulin secretion(GSIS)were detected by radioimmunoassay,mRNA and protein expressions of uncoupling protein 2(UCP2)were determined by RT-PCR and Western blot,respectively.The intracellular ATP content was measured by luciferase bioluminescence.The mitochondrial membrane potential and cellular Ca~(2+) concentration were detected by flow cytometry.Results (1) Various concentrations of Ang Ⅱ had no significant influence on the basal insulin secrection of RIN-m cell(F=0.644,P = 0.634).Except for 0.1 nmol/L AngⅡ,the other concentrations of Ang Ⅱ markedly reduced GSIS of RIN-m cells(F= 118.528,P = 0.000).(2) Compared with the control group,Ang Ⅱ significantly increased mRNA and protein expression of UCP2(F= 1 370,P = 0.000;F=675.175,P = 0.000).(3)Except for 0.1 nmol/L Ang Ⅱ,the other concentrations of Ang Ⅱ significantly decreased the mitochondrial membrane potential,cellular ATP content,and cellular Ca2+ concentration of RIN-m cell(F=4.035,P=0.008;F=3.353,P = 0.013;F=5.867,P = 0.001).Conclusion Ang Ⅱ impairs GSIS of p-cell,the mechanism of impairment may be interpreted that Ang Ⅱ can increase the expression of UCP2,furthermore,it can reduce mitochondrial membrane potential,decrease the content of cellular ATP and the concentration of cellular Ca~(2+),can finally impair the function of β-cell.

Objective:To study the effect of the Shengmai injection combined with norepinephrine on cellular immunity in rats with multiple organ dysfunction syndrome .Methods:40 healthy male rats were randomly divided into the control group ,the Shengmai injection group ,the norepinephrine group and the Shengmai injection combined with norepinephrine group of 10 rats in each group after MODS rat model were established by intraperitoneal injection of zymosan .The plasma level of TNF-α, IL-1β, IL-12 and IL-2 were detected by ELISA ,and the protein expression of caspase-3,caspase-9 and caspase-8 in spleen were tested by IHC during 6 h,24 h,48 h.And the lymphocyte number of apoptosis and the ratio of Th 1/Th2 were tested by flow cytometry.Results: ① Compared with the control group, the plasma concentrations of TNF and IL-1βwere not significant increased in the Shengmai injection group , the norepinephrine group and the Shengmai injection combined with norepinephrine group ,as the extension of treatment time ,the plasma concentration of TNF and IL-1βwere significant lower in treatment groups than control group .While the plasma concentration of IL-12 and IL-2 were significantly increased in treatment groups , and the combined treatment group was more obvious .② Compared with control group,the protein expression of caspase-3,caspase-9 and caspase-8 in spleen were decreased in the Shengmai injection group , the norepinephrine group and the Shengmai injection combined with norepinephrine group ,and the combined treatment group was more obvious.③The lymphocyte number of positive apoptosis were significantly decreased , and the Th1/Th2 ratio were increased in the Shengmai injection group ,the norepinephrine group and the Shengmai injection combined with norepinephrine group ,and the combined treatment group was more obvious .Conclusion: The Shengmai injection combined with norepinephrine can reduce the inflammatory reaction and improve the immune function in the rat with multiple organ dysfunction .

Objective To isolate and cultivate mouse hepatic progenitor cells (mHPCs) from E14.5 mouse fetal liver in vitro and induce mHPCs differentiation into cholangiocytes.Methods Isolation of mHPCs from mouse fetal liver was based on the cell surface antigen delta-like protein 1/preadipocyte factor 1 (Dlk/Pref-1) by a fluorescence-activated cell sorter (FACS).Then mHPCs isolated were co-cultured with/without mouse embryonic fibroblasts (MEFs) by using Transwell.The cell antigen alpha-fetoprotein (AFP), albumin (ALB) and cytokeratin19 (CK19) expression in freshly isolated DLK1+cells or co-cultured for 4 days and 6 days were observed with immunocytochemical method.Results When co-cultured with MEFs, the division and proliferation were observed in most of DLK1+ cells and grape-like aggregation was formed.Cells began to adhere to growth and began to become spindle-shaped on 4th day.The DLK1+cells isolated freshly by FACS were expressed AFP and low levels of ALB but not expressed CK19.But, these cells expressed CK 19 and weak expression of ALB on 4th day.In addition, the expression of CK19 increased and the expression of ALB almost not detected on 6th day.Conclusions Most of DLK1+ cells, isolated from E14.5 fetal livers by FACS, are proved to be mHPCs.Furthermore, these cells can proliferate quickly and differentiate into cholangiocytes by co-culture with MEFs.