Purpose This subject detected the expression of paper miRNA34a,c-myc gene and protein in adenocarcinoma of lung tissue,adjacent mucosa tissue and normal lung tissue exploring the relationship between the two and the clinical significance.Method The project mainly used the method of real-time fluorescent quantitative PCR (qRT-PCR),Western blot and immunohistochecical to detect the expression of miRNA34a and c-myc in normal tissue and adenocarcinoma of lung respectively.Result (1) lower expression of miRNA34a in lung adenocarcinoma tissue,lower than the adjacent and normal tissues (P ＜ 0.01),higher than the metastatic carcinoma group (P ＜ 0.01).C-myc is highly expressed in lung adenocarcinoma tissue,higher than the adjacent and normal tissues (P ＜0.01),lower than the metastatic carcinoma tissue (P ＜ 0.01).miRNA34a and c-myc beside carcinoma tissues and normal tissues also have significant difference between the statistical significance (P ＜ 0.05).(2) Differentiation of low and high there was no statistically significant difference between patients with lung adenocarcinoma (P ＞ 0.05).there were significant differences in the expression of miRNA34a and c-myc in the tissues of metastatic and non metastatic adnocarcinoma.(3)The positive expression of c-myc group of lung cancer patients was lower than that of the negative group (P ＜ 0.05),low expression of the survival time in miRNA34a group was significantly lower than that in high expression group,there was significant difference (P ＜ 0.01).Conclusion Expression of miRNA34a and c-myc both showed a negative correlation in adenocarcinoma of lung.

Objective To investigate the effect of jejunal infusion of amino acids on secretion of gastrointestinal hormone in healthy dogs.Methods Six healthy adult dogs were treated with jejunal fistulas and femoral vein intubation.Twenty-four hours after the operation,solution of 8 different amino acid monomers (experimental group) or normal saline (control group) were infused into the jejunum of the dogs every 24hours.The levels of cholecystokinin (CCK),motilin,and gastrin in the peripheral plasma were measured using radioimmunoassay at the start of infusion (0 minute),and 30,60,90,and 120 minutes after infusion.Results Compared with the control group,the serum CCK level in the phenylalanine group was significantly higher 30 and 60 minutes after infusion [(1.25 ±0.19) ng/L vs.(0.66 ±0.14) ng/L,(1.23 ±0.12) ng/L vs.(0.80 ± 0.03) ng/L,both P ＜ 0.01],while that in the tryptophan group was significantly higher 30 minutes after infusion [(1.08 ±0.26) ng/L vs.(0.66 ±0.14) rig/L,P ＜0.01].The other measurement results showed no statistically significant differences.Conclusions Jejunal infusion of phenylalanine or tryptophan may stimulate the secretion of gastrointestinal hormone to some extent.Aromatic amino acids (phenylalanine and tryptophan) is more potent in triggering the release of CCK than aliphatic (leucine,isoleucine,and methionine) and charged amino acids (aspartic acid,arginine,and glutamate).The mechanism may be related to the properties of the amino acids.

Objective To construct a prokaryotic plasmid to express the testis development related gene 1 (TDRG1) recombinant protein and obtain anti-TDRG1 mAb by immunizing mice, and to identify the biological properties of the mAb. Methods The coding sequence of TDRG1 was amplified by RT-PCR from normal human testis tissue and cloned into the vector pET21, and then was expressed in the E. coli BL 21(DE3) to get TDRG1 recombinant protein. The purified TDRG1 recombinant protein was injected to immunize the BALB/C mice to develop anti-TDRG1 mAb. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted anti-TDRG1 mAb were subcloned with limited dilution. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate titers and subtypes of mAb. Western blot and immunohistochemistry were used to detect specificity of mAb.Results The prokaryotic plasmid expressing the recombinant protein was constructed, and the TDRG1 recombinant protein was expressed and purified. Two hybridoma cell lines that secreted anti-TDRG1 mAb were obtained. The titer of the mAb in ascites was 1∶1.6×10~6, and the subtype of the mAb was IgG_1. Westem blot and immunohistochemistry analysis indicated the mAb showed specific combination with TDRG1 protein in human testes.Conclusion The TDRG1 recombinant protein is highly purified and has strong antigenicity. The anti-TDRG1 mAb is produced successfully.

Objective To explore the clinical efficacy of modified single-portlaparoscopic radical prostatec-tomy(LRP) by atransperitoneal or extraperitoneal approach, and the impact of either approach on immune function in patients with prostate neoplasms. Methods The clinical data on 39 patients who had undergoneextraperitoneal LRP(group A) and 20 patients who had receivedtranperitoneal LRP (group B) in our department from January 2012 to December 2015 were retrospectivelyanalyzed. The prostate neoplasms were preoperatively diagnosed as cancer by ultrasound, CT or MRI, and pathological examinations. The clinical stage was T1-T2c , N0M0 in all patients.The efficacy was assessed by surgical duration, intraoperative blood loss, timeto intestinal function recovery, and post-operative hospital stay in both groups. The indicators for immune function including prostate specific antigen (TPSA and FPSA), immunoglobulin (IgG, IgA, lgM, C3, and C4) and T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) in patientswith prostate neoplasms before and after surgery were detected. Results 59 patients were operat-ed successfully, without converting to open approach. The mean surgical duration, blood loss, time to intestinal function recovery, and postoperative hospital stay were (133.8 ± 68.6) min, (75.6 ± 51.3) mL, (2.2 ± 0.7) days, and (14.7 ± 3.6) days in group A, while (159.4 ± 78.1) min, (102.2 ± 70.8) mL, (2.9 ± 1.1) days, and (15.2 ± 4.1) days in group B. There were significant differences between the two groups (P<0.05). After surgery, levels of IgG, IgA, C3, C4, CD3+, CD4+and CD4+/CD8+were significantly higherin group Athan in group B (P < 0.05). There were no significant differences between the two groups in levels of TPSA, FPSA and lgM (P>0.05). Average postoperative follow-up was 15 months (range 3-36) in 36 patients. No recurrence or metastasis was found in all the patients by imaging studies,and no long-term complications were found. Conclusions Extraperitoneal LRP, as compared with tranperitoneal LRP, has clearer vision, fewer effects on abdominal organs, shorter surgical duration, less blood loss, faster postoperative recovery, and better protection of immune function. It is worth popularizing clin-ically.

Objective This is the first study to explore clinical application value of serum Dickkopf-1 (DKK-1) detection in diagnosis of heptocellular carcinoma (HCC) by magnetic solid phase chemiluminescent immunoassay.Methods The level of serum DKK-1 and AFP in 205 cases of HCC,40 cases of liver cirrhosis,and 200 cases of healthy control were quantitatively detected by Magnetic solid phase chemiluminescent immunoassay.The area under ROC curve,sensitivity and specificity of DKK-1 and AFP for diagnosing HCC were calculated.Results The serum level of DKK-1 in HCC group was significantly higher than those of the liver cirrhosis group and healthy control group (P<0.01).DKK-1 maintained diagnostic sensitivity for patients with HCC who were alpha-fetoprotein (AFP) negative (66.3%).ROC curves showed optimum diagnostic cut-off value was 2.4 ng/mL,area under curve (AUC) was 0.822 (95% CI:0.783-0.856),sensitivity 65.9%,and specificity 87.5%).Moreover,measurement of DKK1 and AFP together improved diagnostic accuracy for HCC versus all controls compared with either test alone [AUC 0.915,95%CI:0.886-0.940),sensitivity 81.5 %(P<0.05)].Conclusion Serum DKK-1 detection has an important clinical value for diagnosis of HCC,especially for HCC with AFP negative.The combined detection of serum DKK-1 and AFP can greatly increase sensitivity and accuracy for diagnosing HCC.

Objective To study the practical clinical value of laparoscopic cholecystectomy(LC) combined with other laparoscopic surgery. Methods 206 patients treated by LC combined with other laparoscopic surgery were analysed retrospectively. Results Among the 206 cases of LC combined with other laparoscopic surgery, LC was combined with choledochal exploration in 64 patients, with appendectomy in 29 patients, with fenestration of hepatic cyst in 38 patients, with other hepatic operations in 13 patients, with unroofing of kidney cyst in 15 patients, with release of abdominal adhesions in 31 patients, and with other operative procedures in 16 patients. Laparoscopic operations were successful in 204 patients, and 2 patients were converted to laparotomy. Postoperative bile fistula occurred in 2 patients; 1 healed with conservative therapy, and 1 healed after reoperation. Conclusions Laparoscopic cholecystectomy combined with other laparoscopic surgery can adequately present the advantages of microsurgery by safe, effective and economical management of various abdominal diseases.

AIM: To investigate the mechanism of proliferation and differentiation of human epidermal stem cells cultured in vitro under the influence of compressive stress.METHODS: Epidermal stem cells were isolated by adhering to type IV collagen and were cultured with conditioned medium,then were detected by Powervision~(TM) two-step immunohistochemical method with keratin 19 and cell cycle analysis.The cultured epidermal stem cells transplanted on silica gel membranes,which were put in a new apparatus,was designed to offer cell culture and intermittent compressive stress(4 kPa,6 kPa,8 kPa,10 kPa,12 kPa) for 2 h,3 times a day simultaneously.A week later,cells on silica gel membranes were identified with keratin 19 and 10 by Powervision~(TM) two-step immunohistochemical method.RESULTS: The new apparatus offered cell culture and intermittent compressive stress simultaneously.The isolated and cultured epidermal stem cells were identified with keratin 19 positive and 84.80 percent of them were showed in G_1 period with cell cycle analysis.Cells on silica gel membranes had been subjected intermittent compressive stress above 8 kPa for a week.The number of the cells was increased,which was more than that in control group.However,some cells identified by immunohistochemical staining with keratin 10 positive were detected among the disposed epidermal stem cells.CONCLUSION: The intermittent compressive stress above 8 kPa induces and promotes epidermal stem cells to proliferate and differentiate,indicating that epidermal stem cells respond to mechanical stress,probably is one of their major biological features.

OBJECTIVESTo summarize the experience in the treatment of 112 cases of complex bone nonunion from 1982 to 1999 in our department and introduce the technique of external skeletal fixation.

METHODSThe two fragment ends of all cases were fixed under pressure with half-ring sulcated external skeletal fixator. Those cases complicated by bone defect or limb shortening were operated on with epiphysiotomy to restore the length of the limb in the period of compressive fixation or after the occurrence of bone union according to the condition of complicated infection and the length of the limb shortened.

RESULTSThe nonunion of the 112 cases was united eventually. The infection in 34 cases was eradicated. Bone union in cases without infection took 3 approximately 7 months (average 5.2 months) and in cases with infection took 5 approximately 11 months (average 5.5 months). The length of the limb in 11 cases with bone defect was restored in the same period of compressive external fixation and another 8 cases achieved after bone union. The length between the injured and healthy limbs was balanced.

CONCLUSIONSWhen external skeletal fixation is employed to treat those troublesome cases of bone nonunion, the pins for fixation are inserted in sites far from the lesions and the non-united fragment ends are exposed only in the area without scars. Consequently, there is little interference with the blood circulation and the osteogenic potency of the fragment ends. The sclerotic bone tissue is not excised, the marrow cavity is not chased to be open and the fragment ends are only moderately modified. As a result, the stability of fixation is increased and further shortening of the limb avoided. External skeletal fixation using small pins with cross penetration results in plastic fixation and promotes bone healing. Bone lengthening with epiphysiotomy can restore the balance of the limbs.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Fracture Fixation ; Fracture Healing ; Fractures, Ununited ; Humans ; Male ; Middle Aged

Objective To investigate the clinical effects of the stent-laparoscopy approach in treatment of elderly patients with malignant colorectal obstruction.Methods Clinical data of 55 elderly hospitalized patients with colorectal cancer-induced bowel obstruction in Wenzhou Central Hospital from January 2009 to January 2014 were evaluated retrospectively.All patients were treated with expandable metallic stents for remission of bowel obstruction.Patients were divided to laparoscopy surgeryobservation group (n =14) receiving laparoscopic surgery after decompression and the laparotomy surgery-control group (n=41) receiving conventional open colorectal cancer operation.The intraoperative information,postoperative complications and long-term curative effect were compared between the laparoscopy surgery-observation vs laparotomy surgery-control group.Results During laparoscopy surgery period,two patients operated by laparoscopy surgery were converted to laparotomy surgery.Surgical time was significantly longer in the laparoscopy surgery-observation group than in laparotomy surgery-control group[(178 ± 33) min vs.(145 ± 31) min,t =3.384,P =0.001],and bowel function recovery and postoperative hospital stay were significantly shorter in the laparoscopy surgery-observation group than in laparotomy surgery-control group[3.1 ± 0.9) d vs.(4.3 ± 1.3) d and (7.1±1.3) d vs.(12.6±5.7) d,t=3.193 and t=2.911,P=0.002 and P=0.005].Intraoperative blood loss was less in the laparoscopy surgery-observation group than in the laparotomy surgery-control group[(63 ± 29) ml vs (86 ± 37) ml,t =2.11,P =0.04],with no significant differences in postoperative complications between the two groups (0.0 vs 9.76 ％,x2 =1.47,P=0.225).Fifty-five patients were followed up for 2 years.All patients survived in the laparoscopy surgery-observation group,but one patient died from the cerebrovascular accident in the laparotomy surgery-control group.Conclusions The stent-laparoscopy approach to treat colorectal cancer (CRC) patients with acute colorectal obstruction is a safe,effective,recovery quick and minimally invasive option for elderly patients.Emergency surgery may be converted to a limited operation by this method.Laparoscopic radical surgery with one-stage anastomosis is feasible.

Objective: To explore the international development trends and research hotspots of outdoor activities affecting the progression of children’s myopia, and to provide a reference for researching on effective ways to prevent children’s myopia. Methods: Totally 291 relevant documents included in the "Web of Science" core set database were used as research objects, and CiteSpace software was used for visual analysis. Results: At present, the publications in this field were mainly in the United States(81), China(80), Australia(76), and Singapore(33); the top three research institutions were "Natl Univ Singapore"(29), "Australian Natl Univ"(27), "Capital Med Univ"(25); the main authors were "Saw SM", "Morgan IG", "Mitchell P". The field has been developed on the basis of "Ophthalmology", "Public, Environmental and Occupational Health", and has been integrated into 32 disciplines. The research content included "exploration of high risk factors for the progression of children’s myopia" and "outdoor activities", "intervention in children’s progression of myopia" and "longitudinal tracking of children’s vision development". Randomized clinical trials that longitudinally track the correlation between changes in eyeballs and the progression of myopia and the effects of outdoor activities on the biological characteristics of children’s eyeballs have become a hot topic in this field. Conclusion Research on the effects of outdoor activities on the progression of myopia in children has increased dramatically. The study of increasing outdoor activities to interfere with the progression of myopia in children and the vertical tracking of key factors affecting the biological characteristics of children’s eyeballs have become the current international trends.