Objective To explore the application of self-made guided wire operating-tray,which is designed for percutaneous tranluminal coronary angioplasty (PTCA),in performing coronary intervention (PCI).Methods A total of 40 patients who were planned to receive PCI were randomly divided into the study group (n=20,using guided wire operating-tray) and the control group (n=20,using traditional method to operate guide wire).Under local anesthesia PCI was carried out in all patients of both groups.The frequency of guide wire winding,the degree of surgical sheet contamination within the operation area,and the comfort and convenience of operating guide wire were assessed,the results were compared between the two groups.Results The frequency of guide wire winding and the degree of surgical sheet contamination within operation area in the study group were remarkably lower than those in the control group (P＜0.05).The comfort and convenience of operating guide wire in the study group was better than that in the control group (P＜0.05).Conclusion In performing PCI,the use of self-made guided wire operating-tray can effectively avoid guide wire winding and reduce the degree of surgical sheet contamination within the operation area.

Objective To investigate the correlation between the reason of heterotopic ossification after cervical artificial disc replacement and the degeneration of cervical facet joints.Methods From May 2009 to May 2012,133 patients who had undergone cervical artificial disc replacement were included into this study.There were 74 males and 59 females with an average age of 42.63±4.15 years old (range,23-56 years old).There were 109 patients who had undergone single level implant,and 24patients double level implant.Cervical spine A-P and the flexion-extension X-rays were taken to assess the range of motion (ROM)of the surgical level.Degeneration degree of patients' cervical facet joints were evaluated by Park grading standards with the preoperative cervical CT scan images on GE-PACS system.Heterotopic ossification (HO) situation were evaluated by McAfee classification through cervical lateral X-ray film.Patients were divided into two groups (HO group and non-HO group) according to whether heterotopic ossification appeared during the follow-up period.The data were collected before surgery to the latest follow-up.Results The average follow-up time was 2.9 years (range,2.0-4.8 years).HO was detected in 25 patients (18.80％,25/133) at latest follow-up.The ROM of surgical level of patients in HO group was significantly lower than non-HO group (6.8°±3.9° vs 9.1 °±2.4°).In addition,the degeneration degree of cervical facet joints of patients in HO group was significantly higher than that in non-HO group.There was a significant correlation between preoperative cervical facet joint degeneration and the occurrence of HO after CDR (r=0.683,P=0.033).Conclusion There was a significant correlation between preoperative cervical facet joint degeneration and the occurrence of HO after CDR.Patients present HO after surgery have significantly higher degeneration degree of cervical facet joints than patients without HO.Patients with higher degree of HO may have a higher level of cervical facet joints degeneration.

Objective cyclin D1 gene plays a significant role in regulating cell cycle progression.Suppression of cyclin D1 protcin expression can effect on cellular proliferation,distribution of cell cycle and apoptosis.This study was to determine whether this effect also existed in chronic leukemia ceil line K562 by inhibiting the expression of cyclin D1 protein through RNA interference in vetro.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan,cyclin D1 protein was examined by using Western blot analysis.Inhibition of cellular proliferation was evaluated hy soft agar colony formation assay.The cell cycle and apoptosis were determined by flow cytometry.Results Expression of cyclin D1 protein was markedly down-regulated and capability of colony formation was suppressed after transfection with pshRNA-419 and pshRNA-575 at 48h.Down-regulation of cyclin D1 protein could effect on distribution of cell cycle arrested at G_0/G_1 phase and markedly induce apoptosis of K562 cells.But there had no above biological effects ob- served after transfection with blank vector and control vector of m-pshRNA-790.Conclusion Down-regulation of cyclin D1 expression can inhibit growth of K562 cells,and effect on distribution of cell cycle arrested at G_0/G_1 phase.The primary results suggest that cyclin D1 gene might serve as an effective target for the treatment of leukemia.

Aged ; Carcinoma, Squamous Cell ; pathology ; surgery ; Cholecystectomy ; Gallbladder Neoplasms ; pathology ; surgery ; Humans ; Male

A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Salvia miltiorrhiza ; enzymology ; genetics

Objective To investigate dental caries in 3-year-old children in Jing′an District of Shanghai and analyze the influence of oral cleaning behavior on caries in children. Methods Six hundread 3-year-old children from 12 kindergartens in Jing′an District, Shanghai were randomly selected by cluster sampling method for oral examination, and a self-administered questionnaire survey on children′s oral cleaning behavior was completed by parents.SPSS 22.0 software package was used for statistical analysis. Results Among the 600 children, the prevalence of dental caries was 42.50%, and the average dmft index was 1.76±2.92.Children who did not brush their teeth had a statistically significant increase in rate of dental caries(χ²=29.995, P < 0.001).Children who brushed their teeth for more than three minutes each time had a statistically significant lower rate of caries(χ²=3.918, P=0.048).Children whose parents helped their children brush their teeth every day had a lower rate of caries than those who brushed teeth by themselves, and the difference was statistically significant (χ²=8.718, P=0.013).Logistic regression analysis showed that age of beginning brushing teeth, times of brushing teeth per day and length of brushing teeth per time were all influence factors for caries in children (P < 0.05). Conclusion For the prevention and control of dental caries in 3-year-old, proper oral cleaning behavior is very important.We should make parents as a priority group for doing oral hygiene in children.Schools should strengthen oral health education for parents.Medical institutions should guide and monitor the effectiveness of oral hygiene in children.It is important to reduce the rate of dental caries in preschool children from three aspects:family, school and medical institutions.

OBJECTIVETo observe the anti-inflammatory effect of total saponins of Panax japonicus on LPS-induced RAW264. 7 macrophages.

METHODThe effect of total saponins of P. japonicus of different concentrations on RAW264. 7 cell viability was determined with the MTT method. The NO kit assay was adopted to detect the NO release of total saponins of P. japonicus to LPS-induced RAW264. 7 cells. The enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). The reverse transeriptase-polymerase chain reaction (RT-PCR) was used to determine the expression of inducible nitric oxide synthase (iNOS) ,TNF-alpha,IL-1beta. The protein expression of nuclear transcription factor-kappaB p65 (NF-kappaB p65) was tested by Western blot.

RESULTThe safe medication range of total saponins of P. japonicus was less than 80 mg x L(-1). Compared with the LPS model group, total saponins of P. japonicus high, middle and low dose groups (0.1, 1, 10, 40 mg x L(-1)) could significantly reduce the secretion of NO, TNF-alpha, IL-1beta of LPS-induced RAW264. 7 cells, and inhibit the expressions of iNOS, TNF-alpha and IL-1beta mRNA and the protein expression of NF-kappaB p65.

CONCLUSIONThis study preliminarily proves the protective effect of total saponins of P. japonicus on LPS-induced RAW264.7 macrophages. Its action mechanism may be related to NF-kappaB signal pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; Interleukin-1beta ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Nitric Oxide ; immunology ; Nitric Oxide Synthase Type II ; genetics ; immunology ; Panax ; chemistry ; Protective Agents ; pharmacology ; Saponins ; pharmacology

Objectives To understand the effects of different doses of vitamin D supplementation on serum calcium,phosphorus,alkaline phosphatase and 25-hydroxyvitamin D levels in very low birth weight infants (VLBWI) and to provide guidance for early prevention of metabolic bone disease in VLBWI.Methods A total of 90 VLBWI hospitalized in the Department of Neonatology,Huzhou Maternal and Child Healthcare Hospital between January 2014 and January 2016 were enrolled and randomly divided into two groups:highdose group and low dose group.High-dose group was given vitamin D 900 U/d orally and low-dose group was given 400 U/d since the eighth day after birth.Serum calcium,phosphorus and alkaline phosphatase levels were detected at 1,7,21 and 42 days of age and serum 25-hydroxyvitamin D levels were detected at 7,21and 42 days of age.Two-sample t-test,Chi-square test,one-way analysis of variance and LSD or Dunnett's T3 test were used for statistical analysis.Results No significant differences in serum calcium,phosphorus and alkaline phosphatase levels were found between the two groups at 1 and 7 days of age,nor were found in serum 25-hydroxyvitamin D level at 7 days of age (all P＞0.05).At 21 days of age,high dose group had higher serum calcium,phosphorus and 25-hydroxyvitamin D levels than low-dose group [(2.38 ± 0.09) vs (2.04 ± 0.15) mmol/L,t=2.421;(1.80±0.50) vs (1.71 ±0.60) mmol/L,t-0.637;(45.58± 18.43) vs (42.53± 16.33) nmol/L,t=0.421],but lower alkaline phosphatase level [(505.12± 185.61) vs (588.32± 168.72) U/L,t=5.314] (all P＜0.05).The same trends were found at 42 days of age.In high-dose group,serum calcium level increased at 7,21 and 42 days of age as compared with that at 1 day of age [(2.43±0.13),(2.38±0.09),(2.39±0.08) vs (2.06±0.57) mmol/L];serum phosphorus level at 7 days of age was lower than that at 1,21 and 42 days of age [(1.31 ±0.26) vs (1.89±0.39),(1.80±0.50),(1.98±0.30) mmol/L];serum alkaline phosphatase level at 7,21 and 42 days of age was higher than that at 1 day of age [(475.18± 133.73),(505.12± 185.61),(538.43 ± 168.16) vs (296.15 ± 99.41) U/L] and a significant increase was observed at 42 days of age as compared with that at 7 days of age;serum 25 hydroxyvitamin D level at 21 days of age was higher than that at 7 days of age,and that at 42 days of age was higher than that at 7 and 21 days of age [(73.55±23.65) vs (30.63± 12.66) and (45.58 ± 18.43) nmol/L];the differences were all statistically significant (LSD or Dunnett's T3 test,all P＜0.05).Conclusions Vitamin D supplementation from the eighth day after birth can improve calcium and phosphorus metabolism in VLBWI and the dose of 900 U/d is more effective than 400 U/d.

ObjectiveTo investigate the effect of Glutamine (Gln) on the expression of PCNA in intestinal tissue of neo-natal rats with necrotizing enterocolitis (NEC), and to explore the protective mechanism of Gln in intestinal mucosa.Methods Forty-eight neonatal rats at the age of 48 hours were selected, and divided into 4 groups, control group, Gln group, NEC group, NECGln group. Each group had 12 rats. Control group were fed mice milk substitutes; Gln group were fed mice milk substitutes mixed with Gln; NEC group were fed mice milk substitutes and had cold/ hypoxia exposure twice a day for 3 days; NECGln group were exposed to cold stress, hypoxia and treated with Gln mixed in the milk. The expression of PCNA was detected using immunohistochemical method.Results Compared with control group were and Gln group, the general condition was worse, and the weight was decreased in NEC and NECGln group. The inifltrated inlfammatory cells, congestion, edema, intrinsic layer separation were observed in intestinal mucosa in NEC and NECGln group. The intestinal villus was lost in severe in NEC and NECGln group. The PCNA index was 34.17±5.78, 34.42±5.38, 15.00±1.94, 30.67±3.14 in control, Gln, NEC and NECGln group respectively, with signiifcant difference between each groups (H=24.32,P=0.000). The expression of PCNA in NEC group was lower than that in normal, Gln, and NECGln group (P<0.008). The expression of PCNA had no signiifcant difference among normal, Gln, and NECGln group (P>0.008).Conclusions The expression of PCNA in intestinal mucosa was decreased in NEC rats. Gln supplement could raise the expression of PCNA in intestinal mucosa of NEC rats, and accelerate the speed of intestinal mucosa repair.