Objective To investigate the effect of recombinant chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia trachomatis(Ct) after Vp1 was co-cultured with Ct (reference strains and clinical strains).Methods The recombinant chlamydiaphage phiCPG1 capsid protein Vp1 was expressed and purified.Equal amount of Ct standard strains (E/UW-5/Cx and D/UW-3/Cx) or clinical strains,which had been incubated with Vp1 protein at the concentration of 53 μg/ml for 3 h at room temperature,were inoculated into McCoy.After cell culture,idione stain and transmission electron microscope were used to observe the effect of Vp1 on the Ct.The effect of Vp1 protein on the cell line McCoy was determined by MTT assay,the responses of Escherichia coli BL21 and DH5α toward Vp1 protein were determined using broth microdilution assays.Results Vp1 had obviously inhibitive effect on Ct,the inhibition ratios were about 40％-70％in clinical strains,72％ in reference strain D and 78％ in E,respectively.Abnormally enlarged RBs were observed after Vp1-treatment and Vp1 could arrest chlamydial developmental cycle using electron microscope.There was no effect of Vp1 on McCoy cells or bacteria BL21 or DH5α.Conclusion The recombinant Vp1 from phiCPG1 has obviously inhibitive effect on the growth of Ct,it will be helpful for the treatment of Ct infection in clinic.

BACKGROUNDDisequilibrium of Th1/Th2 is known as an important cause of allergic asthma with a biased Th2 type response. It has been shown that lipopolysaccharide (LPS) administration during post-sensitization modified the inflammation of asthma via upregulating the Th1 response that decrease the Th2 immunity. We would like to know if, during pre-sensitization, the elevated Th1 response is necessary for LPS exposure to modify the asthmatic response.

METHODSDuring pre- or post-sensitization, 40 microg LPS were intraperitoneal injected (i.p.) to asthmatic mice sensitized and challenged by Dermatophagoides farinae (D. farinea). Inflammation was assessed by examining bronchoalveolar lavage fluid (BALF) for the number and identity of cells and by cytokine titers measured by ELISA. Semi-quantified RT-PCR was used to evaluate the level of Toll-like receptor 4 (TLR4) mRNA in dendritic cells (DCs) from bone marrow (BMDCs).

RESULTSThese investigations demonstrated that LPS exposure during pre-sensitization inhibited the Th2 cytokine and inflammatory infiltration, the same as with LPS exposure during post-sensitization in allergic asthma mice. Contrary to post-sensitization LPS exposure, the Th1 cytokines were not upregulated by pre-sensitization with LPS. Finally, the study failed to show any significant difference between TLR4 mRNA expressed in BMDCs with the two times of LPS exposure.

CONCLUSIONSOur data suggest that elevated Th1 immunity is not required for the modification of the Th2 response induced by LPS exposure during pre-sensitization in asthmatic mice and that pre-sensitization differs from post-sensitization. Immune modulation with treatment is independent of TLR4 expression in BMDCs. This study implicates a potential way to protect from allergic disease and an inflammatory response.

Animals ; Asthma ; chemically induced ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Cytokines ; immunology ; metabolism ; Dendritic Cells ; immunology ; Dermatophagoides farinae ; immunology ; Female ; Lipopolysaccharides ; immunology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Toll-Like Receptor 4 ; genetics

China is the cradle of Chinese herb medicines,with rich plant resources. However, traditional processing methods have many disadvantages, such as high comsumption of organic solvent, long extraction time and high loss of effective constituents. For the purpose of rational use of Chinese herb medicines and accurate analysis on their constituents,the sample pre-treatment method with magnetic nanoparticles as the carrier brought new opportunities in recent years. after consulting literatures,the essay summarizes traditional extraction methods of Chinese herb medicines, characteristics of magnetic materials and their application in the analysis on Chinese herb medicines.
Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnetics ; methods ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry

Objective To construct RhoA siRNA plasmid expression vector.Methods According to the computer aided design,RhoA-specific siRNA gene was synthesized and cloned into the RNAi-Ready Pgenesil-1 Vector.The constructed RhoA-RNAi plasmid were transfected into human HEPG2 cell.Western blot was used to detect the effect of RhoA-RNAi plasmid.Results The recombinant was cloned and the se- quence was obtained.RhoA-RNAi plasmid can down-regulate the expression of RhoA in human hepatocel- lular carcinoma cell line HEPG2.Conclusion Successfully cloning the recombinant makes it possible for searching new mechanism of RhoA in hepatocellular carcinoma.

OBJECTIVETo investigate the method of harvesting of superficial cervical artery island skin flap, and its clinical application in the repair of the maxillofacial and cervical scars.

METHODSThe origin, course, branches and distribution of the superficial cervical artery were studied in 10 adult cadavers. The superficial cervical artery could be divided into three segments: i.e. the segment before entering the trapezius muscle, the segment in the trapezius muscle, and the segment emerging from the trapezius muscle. Fourteen patients with maxillofacial and cervical scar contracture were enrolled in the study. The postburn scars were removed, and the wounds were covered by superficial cervical artery island skin flaps. The survival of the flap after the operation and the recovery of the neck movements were observed.

RESULTSThe length of superficial cervical artery before entering the trapezius muscle was 5.1 +/- 0.4 cm, while that inside the trapezius muscle was 2.1 +/- 0.5 cm, and that after emerging from the trapezius muscle was 4.7 +/- 0.7 cm, respectively. The internal perforating branch was located 7.3 +/- 0.6 cm beside the seventh cervical vertebrae, and 3.9 +/- 0.7 cm above the spine of scapula. The flap was from 16 cm x 7 cm to 35 cm x 12 cm in size. All the flaps survived, except one with partial necrosis in the distal part (3.0 cm x 1.5 cm) of the flap. Follow-up observation for 4 months to 3 years showed satisfactory results.

CONCLUSIONSuperficial cervical artery island skin flap is simple in the flap preparation, and there is no necessity to cut the pedicle, thus it is particularly suitable for the reconstruction of severe postburn maxillofacial and cervical contracture.

Adolescent ; Adult ; Carotid Arteries ; anatomy & histology ; Child ; Cicatrix ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Maxillofacial Injuries ; surgery ; Neck Injuries ; surgery ; Reconstructive Surgical Procedures ; methods ; Scalp ; surgery ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply

OBJECTIVETo screen and identify differentially expressed genes between a fertile patient and another infertile patient who belonged to a large Chinese pedigree affected with androgen insensitivity syndrome (AIS).

METHODSWe constructed the forward and reversed subtracted libraries using genital skin fibroblasts (GSF), which were obtained from the fertile patient MJ and infertile patient ZGJ, as tester respectively. Candidate clones were screened with colony in situ hybridization, dot blot, and Southern blot analysis step by step and conformed with Northern blot analysis. The potential positive clones were sequenced and the homology of the sequences was analyzed.

RESULTSThe forward and reversed subtracted libraries containing differentially expressed pattern of two GSF cell lines were constructed. Two positive clones identified by Northern blot were obtained in the reversed subtracted library. Eleven candidate clones from the two libraries that failed to hybridize with both RNA populations were obtained simultaneously, which might represent differentially expressed low abundance transcripts. Sequencing results and homology analysis demonstrated that the two positive clones were significantly homologous with the genes of autotaxin-t and calcium binding protein calcyclin (S100A6), respectively.

CONCLUSIONSTwo positive clones and eleven clones showing no hybridization signals may represent differentially expressed genes between the two GSFs. This finding may be useful to elucidate the molecular mechanisms leading to phenotypic variation and preserved fertility of the AIS pedigree.

Androgen-Insensitivity Syndrome ; complications ; genetics ; Blotting, Northern ; Fertility ; genetics ; Fibroblasts ; cytology ; Gene Expression Profiling ; Gene Library ; Genitalia, Male ; cytology ; Humans ; In Vitro Techniques ; Infertility, Male ; etiology ; genetics ; Male ; Nucleic Acid Hybridization ; methods ; Pedigree ; Polymerase Chain Reaction ; Skin ; cytology

objecfive To know and compare the intelligence level of children born in different time periods in regions with iodine deficiency disorders(IDD)in Liaoning province.Methods All 7-14 year-old children from ten schools were chosen as the subjects respectively from six villages in each of the six counties and in regions with iodine deficiency,who were respectively born at the initialization of iodinated salt supplying period(1978-1980);non-iodinated salt supplying period(1981-1990);recovery of supplied iodized salt period(1991-1995);universal iodized salt period(1996-2000),respectively.Intelligence quotient(IQ)was measured by Combined Ravens Test in China(CRT-C)and Combined Ravens Test-the Rural,in China,2nd edition(CRT-RC2).Results IQ of children during the non-iodized salt period(91.9±14.3)was significantly lower than the initial supply of iodized salt period(95.8±14.6,q=8.60,P<0.01),recovery of supplied iodized salt period(99.7±14.7)was significantly higher than the initial supply of iodized salt period, non-iodized salt sales period(q = 9.53, 18.13, all P < 0.01 ),universal salt iodization( 104.3 ± 14.9) was significantly higher than the initial supply of iodized salt period, non-iodized salt sales period, recovery of supplied salt iodization(q = 20.00,28.00,10.46, all P < 0.01). Children's rate of mental retardation (IQ≤69) was higher in non-iodinated salt supplying period (6.7%, 88/1314 ) than the initial supply of iodized salt (4.4%, 21/471, χ2 = 3.85, P < 0.05), recovery of supplied iodized salt period(3.3%,48/1470) was significantly lower than non-iodinzed salt supplying period (χ2 = 15.37, P < 0.01), universal salt iodization period(2.7%, 36/1344) was lower than the initial supply of iodized salt period(χ2 = 4.41, P < 0.05) and non-iodinzed salt supplying period(χ2 = 26.34, P < 0.01 ). The IQ and intelligent retarded rates in children born during the initial years of iodinated salt supplying period were not different. The IQ of the children during ten years of non-iodized salt supplying period fluctuated in a "∪" curve, while the intelligent retardation rates in a "∩" curve.The children born during the period of recovery supplied iodized salt increased their IQ and lowered the retardation rates year after year. The IQ of the children in universal iodized salt period kept on increasing while intelligent retarded rates reduced to the lowest level. Conclusions The intelligence level of children born in regions with IDD during non-iodized salt supplying period is remarkably lower than that of the beginning years of iodinated salt supplying period. The intelligence level of children born after universal iodized salt period is remarkably higher than that of the initial iodinated salt supplying period and recovery of supplied iodized salt period, respectively.

Objectives:The purpose of this study was to explore the relationship between the of pericardial adipose tissue (PAT) volume and cardiovascular disease risk factors and energy consumption. Methods:From September 2014 to December 2015,we enrolled 152 inpatients underwent coronary artery CT examination due to suspected coronary heart disease in the Hospital on Integration of Chinese and Western Medicine Affiliated to Nanjing University of Chinese Medicine.The volume of PAT was assessment by 64-slice CT, the energy consumption was assessment by the International Physical Activity Questionnaire. The clinical data, biochemical indexes, PAT volume were analyzed. Results : (1) PAT volume was significantly correlated with age, gender, BMI, HOMA-IR, FPG, HDL-C, TG, total physical activity energy consumption, total middle intensity physical activity energy consumption, total energy consumption of sedentary (P<0.05 or P<0.001); (2) PAT volume of patients with insulin resistance was significantly higher than those without insulin resistance; (3) Multiple regression analysis revealed that age, gender, BMI, total middle intensity physical activity energy consumption were independent predictors of PAT volume. Conclusions: Pericardial adipose tissue (PAT) volume is associated with age, gender, systolic blood pressure, BMI, total middle intensity physical activity energy consumption.

Objective To express the recombinant IL-37b protein and to investigate its role in the regulation of immune responses. Methods The prokaryotic expression vector pET28/IL-37b was constructed. The recombinant IL-37b protein was induced to be expressed and was purified using Ni2+-NTA gel column. C57BL/6 J mice were treated with IL-37b and injected with lipopolysaccharide(LPS)to induce septic shock,and the expression levels of IL-1β,IL-6,IFN-γ in the serum of the mice were detected. Dendritic cells from bone marrow of the mice were isolated and cultured, and were treated with IL-37b. After LPS-induced activation, the expression levels of marker molecules such as CD40, CD80 and MHCII on the cell surface, and cytokines such as IL-1β, IL-10, IL-12 and TNF-α in the culture supernatants were detected by flow cytometry. The CD4 +T cells from mice were isolated and the inhibitory effects of IL-37b on the expression of IFN-γ,TNF-α and IL-10 in the culture supernatants of T cells after induction by LPS were detected. Results IL-37b reduced the expression levels of proinflammatory cytokines in the serum of septic shock mice. IL-37b also inhibited the expression of co-stimulatory molecules and proinflammatory cytokines of the mouse dendritic cells, and suppress the activation of CD4 +T cells in vitro. Conclusions Purified recombinant IL-37b protein has high bioactivity, and can alleviate septic shock in organisms through inhibiting the activation of dendritic cells and related T cell immune responses.

Objective To explore the effects of magnesium sulfate in combination with astragalus membranaceus injection on the birth outcomes and expression of placenta tissue related gene of patients with gestational hypertension. Methods Seventy-six gestational hypertension patients were selected and randomly assigned to control group (n = 38) and treatment group (n = 38). The control group was given intravenous injection of 25％ magnesium sulfate (60 mL) once daily for 7 days,while the treatment group was give intravenous injection of astragalus (60 mL) on the basis of the control group once daily for 7 days. The changes of blood pressure,mean arterial pressure (MAP) and 24 h urine protein content of the two groups were compared. The placenta tissue of the two groups were collected after childbirth. Results After treatment,the total effective rates of the control group and the treatment group were86. 84％ (33/38) and 97. 37％ (37/38) ,respectively; the difference was statistically significant (P < 0. 05). There were significant differences between the control group and the treatment group in systolic blood pressure [(147. 21 ± 20. 01) mmHg vs (128. 46 ± 18. 43) mmHg],diastolic blood pressure [(90. 25 ± 15. 46) mmHg vs (73. 14 ± 14. 53) mmHg],MAP [(126. 76 ± 9. 65) mmHg vs (108. 15 ± 9. 57) mmHg] and the 24 h urine protein content [(2. 65 ±0. 87) g vs (1. 34 ±0. 79) g](P < 0. 05). Conclusion Magnesium sulfate combined with astragalus membranaceus injection can reduce the blood pressure of gestational hypertension patients,improve the pregnancy outcome; the action mechanism maybe related to the up-regulation of PLGF and MMP-9 protein expression of placenta tissue.