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.

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 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.

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.

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.

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 the features of chronic prostatitis during puberty(CPP)and the effects of pelvic floor biofeedback therapy.Methods Totally,25 CPP children (mean age,16 years) and 15 children (mean age,16 years) with normal lower urinary tract as controls were included.In CPP group,NIH-CPSI scores were evaluated,expressed prostatic secretions (EPS) were examined,and bacterial culture was done;and CPP patients were categorized based on the definitions of NIH types.In both groups, urodynamic examination was performed,including evaluation of uroflow curve,maximum flow rate (Q_(max)), post-voiding residual urine (PVR),detrusor-sphincter dyssynergia (DSD),maximum detrusor pressure (P_(det,max))and maximum urethral closure pressure (MUCP).CPP patients underwent biofeedback therapy, and 10 weeks later the effects were assessed.Results In CPP group,NIH typing showedⅡ,ⅢA andⅢB in 1,3 and 21 cases,respectively.Before treatment in CPP and control groups,the incidence of staccato voiding (20 cases vs 1 case),DSD (22 cases vs 1 case),Q_(max)(10.7?3.7 vs 15.0?4.3ml/s),PVR (7.7?4.1vs 3.2?2.6ml),P_(det,max)(115.1?33.6vs 76.8?16.6cm H_2O)and MUCP(176.5?45.7 vs 86.2?28.5cm H_2O)all showed significant differences between the 2 groups(P＜0.05).In CPP group,the differences in pain(4.6?2.2 vs 2.1?1.6),urination (7.9?2.0vs 2.2?1.7),life impact (9.4?2.2vs 2.6?2.1)and total scores(22.0?5.2vs 7.0?4.2) of NIH-CPSI and Q_(max)(10.7?3.7 vs 14.9?5.6) between pre-and post-biofeedback were significant (P＜0.05).Conclusions The main type of CPP is categoryⅢB.The primary symptom is voiding disorder,which leads to greater psychological stress in patients.Children with CPP have pelvic floor dysfunctions and multiple abnormal urodynamic param- eters.The short-term effect of biofeedback strategies for CPP is satisfactory.

Objective To discuss the pathogenesis,diagnosis and treatment of delayed traumatic intracerebral hematoma.Methods The clinic data of traumatic delayed intracranial hematoma patients in this hospital were retro- spectively analyzed.According to clinic observation and CT re-examination,47 cases were diagnosed as delayed trau- matic intracranial hematoma(45 cases by operative treatment,and the other 2 by conservative treatment).Results There were 21 cases of recovery,10 cases of slight disability,8 cases of severe disability,8 cases of death.The total mortality rate was 17 %.Conclusion Brain contusion,subarachnoid hemorrhage and skull base fracture were impor- tant factors of DTICH.Fine-observation and prompt CT re-examination offered excellent results for DTICH.

Objective To evaluate the clinical efficacy of multiple stents placement in the management of hilar cholangiocarcinoma,especially in the complex cases of which the hepatic ducts are invaded.Methods Forty-five consecutive patients with hilar cholangiocarcinoma were treated with percutaneous transhepatic placement of two or three self-expandable metallic endoprostheses.The cause of hilar obstructions in these patients were all cholangiocarcinoma,including Bismuth classification type Ⅱ(n 12 ),Ⅲa(n 17),Ⅲb(n 10),and Ⅳ(n 6).Two or 3 stents were placed in the configuration of T,Y or X over the strictures.Results Stent placement with 2 or 3 endoprostheses was successful in all patients.All patients showed significant decrease in serum bilirubin level.The mortality rate within 30 days of stent placement was 2.2%(1/45).The mean survival and stent patency times were 215.3 d(26— 516 d)and 181.5 d(26—473 d),respectively.Conclusion Deploying of multiple metallic stents is an effective method to treat complex hilar cholangiocarcinoma,especially for the cases of which hepatic ducts are invaded:the henatic ducts should be drained as much as nossible.