OBJECTIVETo construct an hTERT promoter-controlled recombinant plasmid HSV-tk, and to investigate its expression in human gastric cancer cells BGC823 and its effect on proliferation of gastric cancer cells BGC823 in vitro.

METHODSRecombinant plasmid pGL3-hTERT-tk and the corresponding reporter plasmid pGL3-hTERT-tk-Luc(+) were constructed by gene engineering. The recombinant plasmids were then transfected into gastric cancer cells BGC823 via PEG-PEI/Fe(3)0(4) magnetic nano-particles. Fluorescence microscope was used to observe the changes of cell morphology and the transfection efficiency. The expression of the target gene in gastric cancer cells BGC823 was detected by immunohistochemistry. MTT assay was used to evaluate the effect of HSV-tk on the proliferation of BGC823 cells. Normal hepatic cells L02 were used as controls in all the experiments.

RESULTSRecombinant plasmid pGL3-hTERT-tk was successfully constructed, and the length was 1100 bp. pGL3-control-tk-Luc(+), pGL3-basic-tk-Luc(+) and pGL3-hTERT-tk-Luc(+) all could effectively transfect BGC823 cells with high telomerase activity, with the transfection rate being(28.1±2.3)%. Immunohistochemistry study showed significant expression of HSV-tk gene in the cytoplasm of BGC823 cells. MTT showed that 4 days after transfection of pGL3-hTERT-tk, the proliferation of BGC823 was inhibited, and the A570 value was(0.254±0.011), significantly lower than that of L02 cells(0.322±0.013) and that of BGC823 transfected by pGL3-basic-tk (0.357±0.014)(all P<0.05).

CONCLUSIONSMagnetic nano-particles can transfect pGL3-hTERT-tk into BGC823 cells and the expression is achieved. PEG-PEI/Fe(3)0(4) magnetic nano-particles can significantly inhibit the proliferation of BGC823 and may become a potential biological agent for gene therapy of gastric cancer.

Cell Line, Tumor ; Cell Proliferation ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Magnetite Nanoparticles ; Plasmids ; genetics ; Promoter Regions, Genetic ; Simplexvirus ; genetics ; Stomach Neoplasms ; pathology ; therapy ; Telomerase ; genetics ; Thymidine Kinase ; genetics ; Transfection

OBJECTIVETo establish the visual patch clamp whole-cell recording technique and study the properties and functional significance of muscarinic receptor-mediated currents in rat medial vestibular nucleus neurons (MVNn).

METHODSBrain slices containing the MVN were prepared from fifteen Wistar rats. By combining infrared differential interference contrast (IR-DIC) technique and CCD-Camera system with visual patch clamp whole-cell recording technique, twenty healthy neurons were located and muscarinic receptor-mediated currents in rat MVNn were observed and analyzed.

RESULTSVisual patch clamp technique can be used to make direct localization and to make sure of active neuron. In MVNn, a comparison of the current-voltage relationships before and during the application of muscarine, which revealed an increase in the slope of the I-V curve and the reversal potential for this response lay at (-88.4 +/- 4.9) mV (x +/- s), indicates that the activation of muscarinic cholinergic receptors leads to a decrease in potassium current. The test in the voltage sensitivity of the muscarine-induced response, which showed that the effect had a linear current-voltage relationship and reversed at (-86.7 +/- 3.5) mV, indicates that the potassium current blocked by muscarine corresponds to the voltage-insensitive leak potassium current.

CONCLUSIONSVisual patch clamp technique, which was considered better than blind patch clamp technique, can improve the success of sealing process. By the analysis of muscarinic receptor-mediated currents, the data provide support that muscarinic cholinergic mechanisms play a prominent role in the modulation of the excitability of MVNn and also offer a new idea for the efficacy of anticholinergic drugs.

Animals ; Electrophysiological Phenomena ; Neurons ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Receptors, Muscarinic ; physiology ; Vestibular Nuclei ; physiology

OBJECTIVE: To study the influence of the number and position of anchor nails on the Bankart repair by the finite element analysis. METHODS: The 3D CT data of shoulder joints including normal adult volunteers were imported into Mimics 10.0 and Geomagic 2012 software to reconstruct the related tissues, and then meshed in Hypermesh 13.0 software to give material attributes. The position and number of MPC constraints were adjusted in Abaqus 6.14 to establish finite element model of shoulder joint under 6 working conditions including A model(normal control model), B model (Bankart damage control model), C model (1 anchor), D model (2 anchors), E model (3 anchors), F model (4 anchors). The humerus external rotation and forward load were added to simulate the fear test of the shoulder joint forward instability. RESULTS: (1)Normally, the inferior glenohumeral ligament complex accompanied by the external rotation of the humerus head has a wrinkle and a significant stress concentration around its anterior and posterior bundle terminations.(2)The stress of the anterior bundle and posterior bundle of the inferior glenohumeral ligament were increased by 52.33% and 45.67% respectively after Bankart's injury.(3)In each model of anchor repair, the stress concentration was obvious at the anchor site, and the stress of anterior and posterior bundle ligaments could be reduced; there were no obvious differences between C, D models and B model(>0.05), and there were significant differences between E, F models and B model(<0.05). CONCLUSIONS One or two anchors can reduce the stress of the anterior and posterior bundles of the inferior glenohumeral ligament in the repair of Bankart's injury, but when the angle of rotation is large, the stress concentration in the anchor position increases significantly, leading to the increases of the risk of failure. The application of three or four anchors can enhance the constraints on the inferior glenohumeral ligament and reduce the stress of the anchor itself through the "load sharing" effect between anchors, so as to ensure the stability of the shoulder joint and provide a mechanical environment for the early rehabilitation of patients.
Biomechanical Phenomena ; Cadaver ; Finite Element Analysis ; Humans ; Joint Instability ; Shoulder Dislocation ; Shoulder Joint

OBJECTIVETo study the effect of overexpression of Smad7 gene on cell proliferation in human bronchial epithelial cell lines.

METHODSHuman bronchial epithelial cell lines, BEP2D and BERP35T2 cells, were cotransfected with the mammalian expression vectors PCISmad7.neo and pMyc-SEAP, the latter was ac-myc cis-acting enhancer element fused with alkaline phosphatase (SEAP) reporter gene. Expression of c-myc, p15 and p21 mRNA was detected by RT-PCR before and after stable transfection of Smad7 into BEP2D and BERP35T2 cells in order to study the regulation of TGF-beta-mediated growth inhibition.

RESULTSAfter BEP2D and BERP35T2 cells transfected with Smad7, the transcriptional activity of c-myc was significantly increased. Smad7 overexpressing cells showed upregulation of c-myc expression and downregulation of p15 and p21 expression, which contributed to the loss of TGF-beta responses in these cells.

CONCLUSIONOverexpression of Smad7 may facilitate cell proliferation by antagonizing TGF-beta-mediated antiproliferative gene responses.

Bronchi ; cytology ; Cell Proliferation ; Cell Transformation, Neoplastic ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p15 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; genetics ; Epithelial Cells ; cytology ; Humans ; Proto-Oncogene Proteins c-myc ; biosynthesis ; genetics ; Signal Transduction ; Smad7 Protein ; biosynthesis ; genetics ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics

BACKGROUNDLaparoscopic dismembered pyeloplasty is technically feasible for ureteropelvic junction (UPJ) obstruction although it is still challenged by its technical difficulty and time-consuming. In this study, we compared the initial results of retroperitoneal laparoscopic pyeloplasty versus a combined laparoscopic dissection and open reconstruction through a small incision in the treatment of UPJ obstruction.

METHODSSixty-four patients with primary UPJ obstruction underwent pyeloplasty: 32 patients underwent laparoscopic procedure and 32 patients underwent open assisted laparoscopic surgery including two steps, ie, laparoscopic dissection of the UPJ transperitoneally and then pyeloplasty via an extended small incision. The demographic data and intraoperative, postoperative and follow-up conditions of patients were compared between the two groups.

RESULTSPreoperative data were comparable in the patients of the two groups. The operative time was shorter (60.9 minutes vs 157.7 minutes, P < 0.0001) and the complication rate was lower (9.4% vs 31.3%, P < 0.05) in the open assisted group than in the laparoscopic group. The estimated blood loss (42.3 ml vs 47.8 ml), time to have normal diet (37.6 hours vs 33.8 hours), and hospital stay (6.7 days vs 6.2 days) were equivalent. The operative success rate was 97% for the open assisted group and 91% for the laparoscopic group.

CONCLUSIONSThe procedure of combined small incision with laparoscopy for UPJ obstruction is technically easy, and the results are promising. It can be used as an alternative to conventional procedures.

Adolescent ; Adult ; Child ; Female ; Humans ; Kidney Pelvis ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Retroperitoneal Space ; Ureteral Obstruction ; surgery ; Urologic Surgical Procedures

OBJECTIVETo investigate the effect and mechanism of BSDW on the model of allergic rhinitis and the model of guinea pigs by histamine shocking in guinea pigs.

METHODUsing the model of allergic rhinitis in guinea pigs caused by 10% TDI, we observed the effect of BSDW on physiological and pathological symptoms of allergic rhinitis in guinea pigs, the effect of the levels of serum IgE and serum and nasal histamine. Using the model of guinea pigs by histamine shocking, we observed the effect of BSDW on physiological symptoms in guinea pigs.

RESULTBSDW significantly relieved the pathological symptoms of allergic rhinitis in guinea pigs, alleviated the hyperplasia of columnar epithelium, decreased the number of monocyte and eosinocyte compared with the model group. It also reduced the levels of serum IgE, and decreased the release of serum and nasal histamine. BSDW significantly prolonged the occurent time of gasping, eclampsia and death caused by shock, reduced the times of gasping in the model of guinea pigs by histamine shocking.

CONCLUSIONBSDW has significant effect against allergy. The mechanism relates to its effects of decreasing the levels of serum IgE and inhibiting the release of serum and nasal histamine.

Administration, Intranasal ; Animals ; Anti-Allergic Agents ; pharmacology ; Asarum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Histamine ; blood ; Immunoglobulin E ; blood ; Lamiaceae ; chemistry ; Male ; Nasal Mucosa ; immunology ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; immunology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

OBJECTIVETo evaluate the expression of three different RASSF1 transcripts and its clinical significance in lung carcinomas.

METHODSThe mRNA expression of RASSF1A, RASSF1B and RASSF1C was detected by RT-PCR in 51 human lung cancer tissues and 51 matched normal tissues.

RESULTS1. The mRNA expression of three RASSF1 transcripts was detectable in all non-cancer tissues. However, high rate of expression loss of RASSF1A and RASSF1B existed in lung cancer tissues, which was 53.2% (2851) and 37.3% (19/51), respectively. RASSF1C was expressed in all of the tumor tissues. 2. Loss or abnormal down-regulation of RASSF1A was positively related with lymph node metastasis and TNM stage (P < 0.05) and 3. RASSF1B and RASSF1C mRNA expression was not correlated with TNM stage, histological type, differentiation grade or smoking index.

CONCLUSIONThere is a significant expression difference among the three RASSF1 transcripts in lung carcinoma. RASSF1A, closely associated with lymph metastasis and TNM stage of lung carcinoma, should be a new tumor suppressor gene.

Chromosome Deletion ; Chromosomes, Human, Pair 3 ; Genes, Tumor Suppressor ; Humans ; Lung Neoplasms ; genetics ; pathology ; Lymphatic Metastasis ; Neoplasm Staging ; RNA, Messenger ; analysis ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo evaluate the chromium (Cr) levels in blood and urine among general population in China between 2009 and 2010, and thereby to analyze its prevalent features.

METHODSFrom year 2009 to 2010, a total of 11 983 subjects of general population aged between 6 and 60 year-old were recruited from 24 districts in 8 provinces in eastern, central and western China mainland, by cluster random sampling method. The information about their living environment and health status were collected by questionnaire, and 11 983 blood samples and 11 853 urine samples were also collected. Inductively coupled plasma mass spectrometry (ICP-MS) was applied to test the Cr level both in blood and urine; and the Cr distribution in blood and urine among groups of population in different ages, genders and districts, were then analyzed.

RESULTSAmong general population in China, the geometric mean (GM) of Cr concentration in blood was 1.19 µg/L, with median at 1.74 µg /L and 95% percentile at 5.59 µg/L. The Cr concentration in blood among males and females were separately 1.18 µg/L and 1.20 µg/L(P > 0.05); while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 1.00, 1.22, 1.01, 1.40, 1.27 and 1.30 µg/L (P < 0.01), respectively; and the figures in populations from eastern, central and western China were 1.00, 1.70 and 1.98 µg/L (P < 0.01), respectively. Among general population, the GM of Cr concentration in urine was 0.53 µg/L, with median was lower than 0.42 µg/L and 95% percentile at 3.53 µg/L. The Cr concentration in urine among males and females were separately 0.52 µg/L and 0.53 µg/L (P > 0.05);while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 0.56, 0.60, 0.52, 0.50, 0.52 and 0.46 µg/L (P < 0.01), respectively;and the figures in populations from eastern, central and western China were 0.58, < 0.42 and 0.60 µg/L (P < 0.01), respectively.

CONCLUSIONThe study reported the Cr levels in blood and urine among general population in China, and thereby provided basic data evidence for the following Cr biological monitoring studies in near future.

Adolescent ; Adult ; Child ; China ; Chromium ; blood ; urine ; Female ; Humans ; Male ; Middle Aged ; Population Surveillance ; Young Adult

Objective: To investigate the feasibility of establishing a model of allograft penile transplantation in adult beagle dogs and explore the conditions for constructing a stable animal model of penis transplant. METHODS: Following the principles of similarity, repeatability, feasibility, applicability, and controllability in the construction of experimental animal models, we compared the major anatomic features of the penis of 20 adult beagle dogs with those of 10 adult men. Using microsurgical techniques, we performed cross-transplantation of the penis in the 20 (10 pairs) beagle dogs and observed the survival rate of the transplanted penises by FK506+MMF+MP immune induction. We compared the relevant indexes with those of the 10 cases of microsurgical replantation of the amputated penis. RESULTS: High similarities but no statistically significant differences were observed in penile anatomic features between the 20 beagle dogs and 10 men. All the 10 cases of cross-transplantation of the penis were successfully completed in the 20 beagle dogs, of which the transplanted glans survived with normal micturition in 12 but developed necrosis in the other 8; the success rate of one-time venous anastomosis was 95.0% (38/40) and that of one-time arterial anastomosis was 87.5% (35/40), with an average vascular anastomosis time of (71.0±9.0) minutes, a mean operation time of (133.0±10.3) minutes, and a mean blood loss of (135.8±41.4) ml. In the 10 cases of penile replantation, the success rate of one-time venous anastomosis was 100% (20/20) and that of one-time arterial anastomosis was 90.0% (18/20), with an average vascular anastomosis time of (65.0±7.9) minutes, a mean operation time of (117.4±10.0) minutes, and a mean blood loss of (85.0±10.8) ml. In the 12 cases of replantation of the amputated penis, the success rate of one-time venous anastomosis was 100% (24/24) and that of one-time arterial anastomosis was 95.8% (23/24), with an average vascular anastomosis time of (79.0±17.6) minutes, a mean operation time of (125.0±20.6) minutes, and a mean blood loss of (140.0±44.3) ml. No statistically significant differences were found in the relevant indexes among the three groups. CONCLUSIONS The anatomic structure of the corpus cavernosum penis of beagle dogs is highly similar to that of men, almost the same in cross-section anatomy. Microsurgical replantation and allograft transplantation of the penis were both successfully performed in beagle dogs, which showed similar operative indexes to those of human penile replantation. The construction of the allograft penile transplantation model in adult beagle dogs is feasible clinically, with the advantages of operability and repeatability.
Adult ; Anastomosis, Surgical ; Animals ; Arteries ; surgery ; Dogs ; Feasibility Studies ; Graft Survival ; Humans ; Male ; Microsurgery ; Models, Animal ; Necrosis ; etiology ; Operative Time ; Penis ; anatomy & histology ; pathology ; transplantation ; Postoperative Complications ; etiology ; Replantation ; Survival Rate ; Urination ; Veins ; surgery

OBJECTIVETo investigate the clinical effect of fast-track surgery combined with Chinese medicine treatment in devascularization operation for cirrhotic esophageal varices.

METHODSSeventy-two patients with cirrhotic esophageal varices were selected from January 2009 to June 2013, and randomly assigned to a conventional group and a fast-track group (fast-track surgery combined with Chinese medicine treatment) using a randomized digital table, 36 cases in each group. Operation and anesthesia recovery time, postoperative hospitalization and quality of life were recorded and compared between groups during the perioperative period.

RESULTSCompared with the conventional group, the fast-track group had longer operation time (253.6±46.4 min vs. 220.6±51.0 min) and anesthesia recovery time (50.5±15.9 min vs. 23.5±9.6 min; P<0.01); less bleeding (311.3±46.8 mL vs. 356.2±57.5 mL; P<0.01) and less transfusion (1932.3±106.9 mL vs. 2045.6±115.4 mL; P<0.01); as well as faster recovery of gastrointestinal function, shorter postoperative hospitalization and higher quality of life. There were no serious postoperative complications and no further bleeding occurred.

CONCLUSIONFast-track surgery combined with Chinese medicine treatment is a safe and feasible approach to accelerate the recovery of patients with cirrhotic portal hypertension in perioperative period of devascularization operation.

Adult ; Aged ; Anesthesia Recovery Period ; Blood Loss, Surgical ; Blood Transfusion ; Chronic Disease ; Esophageal and Gastric Varices ; surgery ; therapy ; Female ; Humans ; Length of Stay ; Liver Cirrhosis ; complications ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Operative Time ; Postoperative Complications ; Postoperative Period ; Quality of Life ; Splenectomy