Aim To study the effect of high salt diet during pregnancy on the development of renal vessels in offspring rats and its mechanism.Methods Natural pregnant Sprague-Dawley rats were randomly divided into high-salt group and control group.The pregnant rats in the high-salt group were given high-salt diet of 8% NaCl content , while the control group normal diet with 1% NaCl content.In both groups, pregnant rats were given normal drinking water.After delivery, all mothers returned to normal diet and all neonatal rats were breast-fed until one month old.The adult male off springs were used as experimental animals.The vessel tone of renal interlobar arteries and electrophysiological behavior of single vascular smooth muscle cells (VSMCs) were detected respectively.Results The contractile response of renal arteries to phenylephrine(Phe) in high-salt group was stronger than that in the control group(P<0.05).The effect of protein kinase C(PKC) non selective blocker GF109203X on Phe-induced contraction in two groups also showed significant difference(P<0.05).In electrophysiology experiments, Phe inhibited high-conductance Ca2+-activated K+ channel(BK channel) currents in renal arteriolar smooth muscle cells in two groups, and the inhibitory effect was more pronounced in high-salt offsprings (P<0.05).GF109203X could eliminate the response of Phe on BK currents in both groups(P>0.05).Conclusions High-salt diet during pregnancy could increase the sensitivity of renal interlobar arterial contractile response to Phe in adult male offsprings, which is associated with PKC-mediated BK channels pathway.Maternal high-salt diet during pregnancy may increase the risk of renal vascular diseases in adult offsprings.

In this paper, optimization of the conditions of microwave technique in extraction process of Guizhi Fuling capsule in the condition of a pilot scale was carried out. First of all, through the single factor experiment investigation of various factors, the overall impact tendency and range of each factor were determined. Secondly, L9 (3(4)) orthogonal test optimization was used, and the contents of gallic acid in liquid, paeoniflorin, benzoic acid, cinnamic acid, benzoyl paeoniflorin, amygdalin of the liquid medicine were detected. The extraction rate and comprehensive evaluation were calculated with the extraction effect, as the judgment basis. Theoptimum extraction process of Guizhi Fuling capsule by microwave technology was as follows: the ratio of liquid to solid was 6: 1 added to drinking water, the microwave power was 6 kW, extraction time was 20 min for 3 times. The process of the three batch of amplification through verification, the results are stable, and compared with conventional water extraction has the advantages of energy saving, time saving, high efficiency advantages. The above results show the optimum extracting technology of high efficiency, stable and feasible.
Capsules ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; Microwaves ; Technology, Pharmaceutical ; methods

B-type natriuretic peptide (BNP) is a plasma marker of left ventricular dysfunction and cardiac volume overload. Currently it is mainly used in the cardiovascular field. BNP is an intrinsic regulator of the embryonic stem cell proliferation, and the reduction in BNP can increase the apoptosis rate. The epitope of N terminal pro-brain natriuretic peptide-BNP is most stable. BNP1-32 has the strongest biological activity but with lower plasma level in heart failure patients. The plasma BNP level plays an important role in the diagnosis, prognosis, hospital admission and mortality of heart failure, and can be used as a monitoring indicator in the treatment of heart failure. The deficiency of corin enzyme in patients with heart failure can cause the increase of cracking pro-BNP. BNP can also provide diagnostic and prognostic information for other populations and diseases. Genetic studies on BNP and its receptors also provide important information. Nesiritide, neutral endopeptidase inhibitors, and vasopeptidase inhibitors of the natriuretic peptide synthesis have been used for the treatment of cardiovascular disorders. However, more reliable and accurate approaches for detecting BNP and N terminal pro-brain natriuretic peptide-BNP require further investigations.
Cardiovascular Diseases ; blood ; diagnosis ; drug therapy ; Humans ; Natriuretic Peptide, Brain ; blood ; physiology ; therapeutic use

Objective To localize the sinoatrial node (SAN) of the newborn rabbits in vivo and cut it for purifying cultivation and study the morophologic characters of primary cultured pacemaker cells of SAN under light microscope and transmissional electron microscope. Methods Hearts of the newborn rabbits were embedded in paraffin for HE-staining and observed the location, form of SAN under optical microscope; SAN cells isolated from neonatal rabbits cultured and purified with the method of differential attachment and BrdU-treatment.Results SAN localized in the anterior wall of the superior vena cava and the posterior-lateral atrial wall.There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinctly different types of cells were observed among the cultured cells of SAN: spindle, araneiform and polygon. The spindle cells covered the greatest proportion of the cultured cells of SAN (59.6%?7.3%). The frequency of spontaneous contraction of spindle cells was the highest among the constrcting cells (145 ?9)time/min. The results of ultrastructure observation showed that myofibrils and other organelles in spindle cells were poorly organized and significantly decreased in number compared with araneiform cells. There was no significant difference between araneiform cells isolated from SAN and from atrial muscle.Conclusion Among the cultured cells from neonatal rabbits SAN, the spindle cells are the pacemaker cells of SAN.

Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.

OBJECTIVETo evaluate the application of side-to-side anastomosis of the lesser curvature of stomach and jejunum in laparoscopic Roux-en-Y gastric bypass (LRYGB).

METHODSClinical data of 29 patients with type 2 diabetes mellitus (T2DM) undergoing side to side anastomosis of the lesser curvature of stomach and jejunum in LRYGB from May 2012 to November 2012 in Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University were analyzed retrospectively.

RESULTSAll the procedures were successfully completed without conversion to laparotomy. The side-to-side anastomosis of the lesser curvature of stomach and jejunum avoided the laparoscopic suture. No gastrojejunostomy anastomotic bleeding, fistula, obstruction and other complications occurred after operation and no complications of gastrojejunostomy anastomosis were found during a follow up of 1 to 7 months.

CONCLUSIONSSide-to-side anastomosis of the lesser curvature of stomach and jejunum in LRYGB can manipulate the size of anastomosis accurately and avoid the laparoscopic suturing. It is simple and easy to learn.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Follow-Up Studies ; Gastric Bypass ; methods ; Humans ; Jejunum ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach ; surgery ; Treatment Outcome

OBJECTIVETo observe the effect of Wenyang Huazhuo Tongluo Recipe (WYHZTLR) on the proportion of T helper 17 cells (Thl7)/regulatory T cells (Treg), and serum levels of IL-17 and IL-10 in peripheral blood of systemic sclerosis (SSc) patients with yang qi insufficiency and turbidity induced collaterals blockage syndrome (YQITICBS).

METHODSTotally 82 SSc patients were randomly assigned to the Western medicine group (as the control group) and the integrated Chinese and Western medicine group (as the treatment group), 41 cases in each group. All patients took methotrexate (MTX) tablet and prednisone tablet. Patients in the treatment group additionally took WYHZTLR. The treatment course for all was six consecutive months. Besides, another 70 healthy volunteers were recruited as a healthy control group (as the healthy group). Percentages of Th17 and Treg in peripheral blood were detected by flow cytometry. Serum levels of IL-17, IL-10, von Willebrand factor (vWF), aminoterminal propeptide of type l procollagen (PIIINP), and cross-linked carboxyterminal telopeptide of type I collagen ( I CTP) were measured by double antibody sandwich ELISA. The correlations between Th17/Treg and levels of vWF, PIIINP, I CTP, skin score, and disease activity index were observed by Pearson correlation analysis.

RESULTSThe percentage of Th17 in peripheral blood, ratios of Th17/Treg, and the serum level of IL-17 were significantly higher, but the percentage of Treg and the serum level of IL-10 were significantly lower in SSc patients, when compared with those of the healthy group (P <0. 05, P <0. 01). Compared with the same group before treatment, the percentage of Thl7, ratios of Thl7/Treg, and levels of IL-17, vWF, and PIIINP all decreased in the two groups after treatment (P <0.05, P <0.01), but the percentage of Treg and the IL-10 level increased (P <0. 05, P <0. 01). Meanwhile,the level of I CTP was higher in the treatment group after treatment (P <0. 05). The improvement of all indices except the percentage of Th17 was more obvious in the treatment group than in the control group (P <0. 05, P <0. 01). The ratio of Th17/Treg was positively correlated with levels of vWF, PIIINP, skin score, and disease activity index before and after treatment respectively (P <0. 01), but with no obvious correlation with the level of I CTP (P >0. 05).

CONCLUSIONWYHZTLR could achieve its therapeutic effect on SSc patients by regulating Th17/Treg imbalance, lowering levels of vWF and PIIINP, and elevating the level of I CTP.

Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-10 ; Interleukin-17 ; Methotrexate ; Peptide Fragments ; Procollagen ; Scleroderma, Systemic ; drug therapy ; T-Lymphocytes, Regulatory ; Th17 Cells

"Wise men could recognize similarities, but the fool only recognizes differences" in Su-wen, which expounded clinical thinking methods of Chinese medicine (CM). "To recognize similarities and differences simultaneously" is of important clinical significance in understanding the laws of diseases. CM pays much attention to recognize similarities, while modern medicine emphasizes the differences observed. In order to develop integrative medicine (IM), similarities recognition and differences identification must be combined together to innovate new thinking methods of IM.
Humans ; Integrative Medicine ; standards ; Medicine, Chinese Traditional

Adult ; Bile Duct Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; pathology ; surgery ; Cholangiocarcinoma ; diagnosis ; metabolism ; pathology ; surgery ; Cholecystectomy ; methods ; Diagnosis, Differential ; Hepatectomy ; methods ; Humans ; In Situ Hybridization ; Keratin-19 ; metabolism ; Keratin-7 ; metabolism ; Keratin-8 ; metabolism ; Magnetic Resonance Imaging ; Male ; RNA, Viral ; metabolism ; Tomography, X-Ray Computed

The purpose of this study was to evaluate whether the DNA vaccine containing idiotypic gene fragment of human B-cell lymphoma cell line Namalwa could elicit the specific anti-idiotypic immune response in vivo. The candidate gene fragment of the lymphoma cell, variable region of heavy chain (VH) of the membranous immunoglobulin, was amplified using Ig superfamily primers by means of RT-PCR. Also, the intact cDNA of murine monocyte chemoattractant protein (MCP-3) was cloned and used as the adjuvant molecular. The two gene fragments of VH and MCP-3 were fused together by 8aa linker peptide with recombinant PCR. Subsequently, the fusion gene fragment was cloned into eukaryonic expression vector pcDNA3.1 to construct DNA vaccine plasmid. Prior to the immunization, the transient transfection coupled with RT-PCR was performed to prove that the recombinant plasmid could express in eukaryonic cells in right way. Then two groups of mice were immunized by intramuscular injection with DNA vaccine and mock plasmid pcDNA3.1 respectively. Three times of injection were performed with 100 micro g plasmid respectively at the beginning of the experiment and 2, 4 weeks after the first injection for all the groups. FACS analysis was chosen to detect the antibodies recognizing lymphoma cells at different time following vaccination. The results demonstrated that specific anti-idiotypic antibody could be detected in the group of DNA vaccine immunized mice as early as eight weeks after the first immunization. Further test demonstrated that the anti-idiotypic antibody could maintain for at least twenty weeks with high titer. Anti-idiotypic antibodies were elicited in three of five mice of the DNA vaccine immunized group. The Abs of DNA vaccine immunized mice could only recognize Namalwa cell line instead of another unrelated human cell line A549. There is no cellular response detected in the DNA vaccine immunized mice. It is concluded that the DNA vaccine containing fused MCP3-VH sequence could elicit specific anti-idiotypic antibody against B-cell lymphoma in vivo and could be used in further study of DNA vaccine against B-cell lymphoma. The results would provide the basis for further studies and optimization of this therapeutic strategy on patients with B-lymphoproliferative disease.
Animals ; Antibodies, Anti-Idiotypic ; blood ; COS Cells ; Cancer Vaccines ; immunology ; Chemokine CCL7 ; Cytokines ; Humans ; Immunization ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Lymphoma, B-Cell ; immunology ; Mice ; Mice, Inbred BALB C ; Monocyte Chemoattractant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; immunology