OBJECTIVETo study the selective cytotoxic effect of lentivirus-mediated double suicide gene (CD/TK) against human gastric carcinoma cells SGC-7901 in vitro.

METHODSSGC-7901 cells were infected with FGW-KDRP-CD/TK vector and the infection efficiency was observed under a fluorescence microscope. The morphological changes of the infected cells were observed by Giemsa staining. Flow cytometry (FCM) was employed for cell cycle analysis, and the expression of CD/TK was detected by RT-PCR. The infected cells were then treated with the prodrugs ganciclovir (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT method.

RESULTSThe infection efficiency of the lentiviral vector in SGC-7901 cells increased with the titer of the virus, which produced no significant effect on the cancer cell morphology in vitro or on the percentages of G0-G1, G2-M and S phase cells (P>0.05). RT-PCR demonstrated the expression of CD/TK gene in SGC-7901 cells infected by FGW-KDRP-CD/TK. The infected cells were highly sensitive to the prodrugs with a dose-dependent cytotoxic effect within a specific concentration range of the drugs, whereas the non-infected cells were not sensitive to the prodrugs. Combined use of the two prodrugs produced an obviously stronger inhibitory effect than either of the them (P<0.05). When combined, GCV and 5-FC at the concentration of 0.1+40, 1+80, 10+160, and 100+320 mg/L demonstrated a synergetic effect with a CDI<1.

CONCLUSIONLentivirus-mediated CD/TK fusion gene system can selectively kill gastric cancer cells, and the two prodrugs show a synergistic cytotoxic effect.

Adenocarcinoma ; genetics ; pathology ; Cell Line, Tumor ; Cytosine Deaminase ; biosynthesis ; genetics ; Cytotoxins ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stomach Neoplasms ; genetics ; pathology ; Thymidine Kinase ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

OBJECTIVETo study the regulatory effect of glucagon-like peptide-1 (GLP-1) on cell proliferation of skeletal myoblast strain L6 and its possible signal mechanism.

METHODSL6 cells cultured in DMEM high glucose culture medium containing 10% FBS were divided into control group (C, without addition), GLP-1 group (G, added with 10 nmol/L GLP-1), PI3K inhibitor group (W, added with 50 nmol/L PI3K specific inhibitor wortmannin), and GLP-1 + PI3K inhibitor group (GW, added with 10 nmol/L GLP-1 and 50 nmol/L wortmannin) according to the random number table. Cell proliferation activity was detected with MTT assay at post culture hour (PCH) 24, 48, 72 (denoted as absorbance value). At PCH 24, the change in cell cycle was evaluated with flow cytometer, the expression level of proliferating cell nuclear antigen (PCNA) was determined with immunohistochemical staining, the protein levels of phosphorylated PI3K (p-PI3K) and p-Akt were determined with Western blotting. Data were processed with multi-group analysis of variance.

RESULTS(1) The cell proliferation activity at PCH 48, 72 in G group was respectively 0.660 +/- 0.120, 0.870 +/- 0.240, all significantly higher than those in C group (0.530 +/- 0.060, 0.700 +/- 0.100, with F value respectively 5.46, 5.90, P < 0.05 or P < 0.01). The cell proliferation activity in W group at each time point was lower than that in C group. The cell proliferation activity in GW group at PCH 48, 72 was respectively 0.510 +/- 0.080, 0.740 +/- 0.160, all lower than those in G group (with F value respectively 5.46, 5.90, P < 0.05 or P < 0.01). (2) The percentage of S phase cell in G group at PCH 24 [(15.7 +/- 0.4)%] was significantly higher than that in C group [(13.6 +/- 0.6)%] and GW group [(10.1 +/- 0.6)%], while that in W group [(6.8 +/- 1.2)%] was lower than that in C group (with F values all equal to 15.39, P values all below 0.01). (3) PCNA level in G group at PCH 24 [(51.24 +/- 1.18)%] was markedly higher than that in C group [(36.72 +/- 1.56)%] and GW group [(25.90 +/- 1.22)%], and while in W group [(21.70 +/- 0.09)%] was lower than that in C group (with F values equal to 783.80, P values all below 0.05). (4) The protein level of p-Akt in G group at PCH 24 was significantly higher than that in the other 3 groups, while that in W group was lower than that in C group (with F values equal to 94.43, P values all below 0.01). There was no obvious difference in protein level of p-PI3K at PCH 24 among G, GW, and C groups ( F = 20.94, P > 0.05). The protein level of p-PI3K at PCH 24 in W group was lower than that in C group (F = 20.94, P < 0.05).

CONCLUSIONSGLP-1 can promote cell proliferation of skeletal myoblast by accelerating the progression of cell cycle and increasing the synthesis of DNA, which can be attributed to PI3K/Akt signal pathway.

Androstadienes ; pharmacology ; Animals ; Cell Cycle ; Cell Line ; Cell Proliferation ; drug effects ; DNA ; biosynthesis ; Glucagon-Like Peptide 1 ; pharmacology ; Myoblasts, Skeletal ; drug effects ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Rats ; Signal Transduction ; drug effects

OBJECTIVETo explore the clinical effect of transplantation of the long head of biceps femoris muscle flap in combination with semi-V posterior thigh fasciocutaneous flap for repair of pressure sores over ischial tuberosity.

METHODSEight patients with 10 deep pressure sores over ischial tuberosity were admitted to the First Affiliated Hospital to the PLA General Hospital and the 98th Hospital of PLA from April 2004 to June 2010. The wounds measured from 2 cm × 2 cm to 6 cm × 4 cm were covered with the long head of biceps femoris muscle flap and semi-V posterior thigh fasciocutaneous flap (ranged from 10 cm × 6 cm to 13 cm × 8 cm). The condition of flaps was observed and followed up for a long time.

RESULTSAll flaps survived. Nine wounds healed by first intention. Subcutaneous accumulation of fluids occurred in one wound with formation of a sinus at drainage site, and it healed after dressing change for 25 days. Patients were followed up for 7 to 34 months. Sore recurred in one patient 9 months after surgery, and it was successfully repaired with the same flap for the second time. Flaps in the other 7 patients appeared satisfactory with soft texture and without ulceration.

CONCLUSIONSThis combined flap is easy in formation and transfer, and it causes little side injury with good resistance against pressure. It is a new method for repair of pressure sore over sacral region.

Adult ; Female ; Humans ; Ischium ; Male ; Middle Aged ; Muscle, Skeletal ; transplantation ; Pressure Ulcer ; surgery ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps

OBJECTIVETo develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.

METHODS16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.

RESULTSAfter PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.

CONCLUSIONThe suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

Bacillus anthracis ; isolation & purification ; Bioterrorism ; prevention & control ; DNA Primers ; DNA, Bacterial ; analysis ; Francisella tularensis ; isolation & purification ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Yersinia pestis ; isolation & purification

OBJECTIVETo investigate the incidence of lymph node metastasis (LNM) in the right para-tracheal triangle (RPT) of esophageal carcinoma patients and the technique of dissection.

METHODSOn the top of double mediastinal and abdominal lymphadenectomy, 333 esophageal carcinoma patients received RPT lymphadenectomy through the right pleural apical approach from 1990 to 2001.

RESULTSIn these 333 patients, the lymph node metastasis (LNM) rate in the RPT was 36.40%. A total of 457 nodes among 2 159 nodes removed gave a metastasis degree of 24.96%. The LNM rates in RPT for cervical, upper third, middle third, and lower third segments of esophagus were 66.67%, 45.45%, 34.19% and 15.79% (P < 0.05), while their respective metastasis degrees were 44.44%, 27.04%, 24.32% and 18.92% (P > 0.05). The frequency of positive nodes in the RPT for PTI, PT1, PT2, PT3 and PT4 was 0, 17.24%, 28.7%, 45.16% and 53.57%, while those of metastasis degree were 0, 8.77%, 17.62%, 33% and 41.17% (P < 0.01). The frequency of LNM in the RPT in papillary, erosive, patch-like and covert type of early tumor was 40%, 3.85%, 0 and 0 (P < 0.05), while those of the metastasis degree were 29.41%, 1.82%, 0 and 0 (P < 0.01). Higher rate of LNM in progressive stenotic esophageal carcinoma was observed compared with those of the other gross types (56.52%, P < 0.05), so was the degree (P < 0.01). The frequency of LNM in the RPT for mono-focal and multi-focal tumor was 34.98% and 70% without significant difference (P > 0.05), while the degree was 24.29% and 53.33% (P < 0.05). Postoperative complications were: leak (0.6%), and recurrent laryngeal nerve injury (1.2%). No injury of vein or infra-clavicular artery, tracheal damage or mortality occurred.

CONCLUSION1. The lymph node metastasis from esophageal carcinoma has a tendency of wide spread and right para-tracheal triangle is an important region to be doomed. 2. With location, depth of tumor invasion and differentiation of tumor as major factors affecting LNM of esophageal carcinoma, dissection of this region should be paid more emphasis. 3. In early lesions, higher frequency of LNM in the RPT is found in papillary and erosive lesions than in the other macroscopic types. 4. Exposing the RPT, lymph node by dissection through a right pleural apical approach is very important and significant.

Adult ; Aged ; Cardia ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Esophagus ; pathology ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Neck ; Neoplasm Invasiveness

OBJECTIVE: To analyze the occupational hazard factors and their critical control points in the process of construction of a large-scale construction project.METHODS: The engineering analysis, worksite survey occupational health and occupational hazard factor testing were conducted in a large-scale construction project, and the results were analyzed. RESULTS: During the process of construction of this large-scale construction project, there were many occupational hazard factors such as silicon dust, welding smoke, wood and other dusts, inorganic compounds of manganese, carbon monoxide, carbon dioxide, nitric oxide, ozone, noise, high temperature, hand-transmitted vibration and ultraviolet irradiation, among them, silicon dust and noise were the most common ones. The over standard rates of exposure concentration of short term of total dust and respirable dust in the workplace were 68.2%(15/22) and 40.9%(9/22), and the over standard rates of exposure concentration of time weighted average were 54.5%(12/22) and 13.6%(3/22), respectively. The over standard rates of the noise intensity of area sampling and personal sampling in workplace were 69.2%(45/65) and 61.0%(25/41) respectively. The four hours energy equivalent frequency-weighted acceleration to vibration of three hand-transmitted vibration positions has been detected, and the result has surpassed the occupational exposure limit.The results of other occupational hazard factors such as high temperature, ultraviolet radiation, wood dust, welding smoke, other dust, manganese inorganic compounds, carbon monoxide, carbon dioxide, nitric oxide, nitrogen dioxide, hydrogen sulfide, hydrogen cyanide and ozone all met the occupational exposure limits. CONCLUSION: There are various occupational hazard factors in the process of construction of this large-scale construction project, among them, noise, dust and hand-transmitted vibration are the most prominent hazards.These hazards are the critical control points of this type of construction projects.

Purpose: This study aims to investigate the current status of affiliate stigma among parents of autistic children, analyze the influencing factors, explore the relationship among mindfulness, coping styles, and affiliate stigma, and verify the mediating role of coping styles between mindfulness and affiliate stigma in parents of children with autism in China.MethodBetween February and April 2023, the Child Development Behaviour Centre of a public hospital in China recruited 345 parents of children with autism. These parents completed the general information questionnaire, the Mindful Attention Awareness Scale, the Affiliate Stigma Scale, and the Simple Coping Style Questionnaire. We then adapted the Hayes Process Macro and Bootstrap methods to examine the mediating effects of coping styles between mindfulness and affiliate stigma. Results: (1) The total affiliate stigma score of parents of children with autism was 48.53 (standard deviation:: 10.74). Parents' age, monthly family income, duration of care, mindfulness, and coping styles were the influencing factors of parental affiliate stigma. (2) Mindfulness was positively correlated with positive coping style (r = 0.33, p < .01) and negatively correlated with negative coping style, affiliate stigma (r = −0.38, −0.39, p < .01), whereas affiliate stigma was negatively correlated with positive coping style (r = −0.34, p < .01) and positively correlated with negative coping style (r = 0.41, p < .01). (3) Positive coping style and negative coping style play a parallel mediating role between mindfulness and affiliate stigma of parents of autistic children. Conclusions Parents of children with autism experience significant levels of affiliate stigma. Mindfulness has a direct impact on associated stigma in parents of children with autism and also indirectly predicts associated stigma through the intermediary influence of positive and negative coping styles. Healthcare professionals could perform mindfulness interventions from an optimistic psychology viewpoint to boost parents' mindfulness and coping abilities, thereby accomplishing the objective of mitigating affiliate stigma.

OBJECTIVETo study the expression of the ID3 protein in prostate cancer and its clinicopathological significance.

METHODSWe detected the expression of the ID3 protein in PC-3M cells by indirect immunofluorescence, and that in 29 prostate cancer and 15 prostate hyperplasia specimens by immunohistochemistry. Then we analyzed the correlation between the expression level of ID3 and the clinicopathological parameters.

RESULTSThe ID3 protein was expressed predominantly in the nucleus of PC-3M cells. Its expression rate was 82.7% (24/29) in the prostate cancer specimens, significantly higher than 6.6% (1/15) in prostate hyperplasia (P < 0.05), and was positively correlated with the Gleason score of prostate cancer (P < 0.05).

CONCLUSIONThe ID3 protein is expressed in prostate cancer, and is elevated with the increase of Gleason score.

Aged ; Aged, 80 and over ; Fluoroimmunoassay ; Humans ; Immunohistochemistry ; Inhibitor of Differentiation Proteins ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the influence of different amount of allogeneic microskin in mixed grafting with certain quantity of autologous microskin on wound healing in rats. Methods Male Wistar rats served as alloskin donor rats. Forty female SD rats with full thickness skin defect were enrolled in the study, and they were randomly divided into four groups, i.e. group I (n=10, with allogeneic microskin graft at area expansion rate of 10:3); group II (n=10, with autologous microskin graft at area expansion rate of 10:1); group III (n=10, with mixed grafting of autologous and allogeneic microskin at area expansion rate of 10:1, respectively); group IV (n=10, with mixed grafting of autologous and allogeneic microskin at area expansion rate of 10:1 and 10:3, respectively). The wound healing rate, wound contraction rate and histological changes were observed at the 2, 3 and 4 post graft weeks (PGW).

RESULTS(1) In group I, there was mainly granulation tissue with some de novo epithelial cells appearing at the wound edge along with the rejection of grafted allogenous skin in the rat wound. In group II, there was still some granulation tissue remaining at 2 PGW due to insufficient amount of microskin. However, the wounds in the mixed grafting group appeared almost totally epithelialized. (2) Various amounts of mononuclear inflammatory cell infiltration and different degrees of angiectasis were observed in the dermal layer after the skin grafting in all groups, especially in group II and IV. There was thickening of the epithelial layer in all groups except group I. (3) The wound healing rate decreased obviously along with the development of rejection in group I at 2 to 4 PGWs. The wound healing rate was (55 +/- 26)% in group II, which was obviously lower than that in group III (88 +/- 6)% and in group IV (76 +/- 10)% at 3 PGWs (P < 0.01). (4) The contraction rate of the wound in group IV (69 +/- 7)% was much higher than that in group I (58 +/- 11)% at 3 PGWs (P < 0.05), and there was no difference among all the other groups.

CONCLUSIONWound healing can be obviously accelerated by mixing some autologous microskin with appropriate amount of alloskin. Moreover, certain amount of autologous microskin (expansion rate 10:1) mixed with the same proportion of allogeneic microskin seems to be more beneficial in promoting wound healing.

Animals ; Disease Models, Animal ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Skin ; injuries ; Skin Transplantation ; methods ; Transplantation, Autologous ; Transplantation, Homologous ; Wound Healing

OBJECTIVETo explore significance of high-risk human papillomavirus (HR-HPV) testing in atypical squamous cells, cannot exclude high grade squamous intraepithelial lesion (ASC-H).

METHODSPresence of HR-HPV DNA was examined in 45 patients with ASC-H using hybrid capture II (HC-II) test. Colposcopic examination and biopsy were taken all results were evaluated.

RESULTSOverall, 33 of 45 (73.3%) ASC-H cases were biopsy proven cervical intraepithelial lesion (CIN). 36 of 45 ASC-H cases were HPV-DNA positive, including 19 cases of HSIL and over lesion; whereas no HSIL or over was found in 9 HR-HPV negative cases. Sensitivity and negativity predictive value of HR-HPV in ASC-H with HSIL and over lesion were both 100%.

CONCLUSIONSASC-H strongly predicts the presence of HSIL, HR-HPV may serve as a predict select whether a patient with ASC-H should take colposcopic examination immediately, patients with positive HR-HPV should undergo immediate colposcopic examination, while negative HR-HPV is an excellent predictor of the absence of HSIL.

Adult ; Aged ; Biopsy ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Colposcopy ; DNA, Viral ; analysis ; Female ; Humans ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; Precancerous Conditions ; pathology ; virology ; Uterine Cervical Neoplasms ; pathology ; virology ; Uterine Cervicitis ; pathology ; virology ; Vaginal Smears ; Young Adult