OBJECTIVETo observe targeted expression of recombinant lentivirus-mediated (Lv)-hTERTp-TK and Lv-hTERTp-tumstatin in HepG2 cells, and explore the inhibitory effect of their combination on HepG2 cells both in vitro and in vivo.

METHODSLv-hTERTp-TK and Lv-hTERTptumstatin were used to infect HepG2 and L02 cells at different MOIs. Transfection efficiency was observed by fluorescence microscopy. Expression of TK and tumstatin mRNA was detected by reverse-transcriptase PCR. Proliferation and apoptosis were detected by MTT and flow cytometry, respectively. The HepG2 cells were examined by real time-PCR and western blotting to determine expression level of bcl-2 and VEGF mRNA and protein.A murine hepatocellular carcinoma model was established by injecting 1 * 10(7) HepG2 cells into 30 BALB/c nude mice. The modeled mice were randomly divided into a control group, mock group, Lv-hTERTp-tumstatin group, Lv-hTERTp-TK group, and combination group for four weeks of injections at regular intervals of PBS, Lv-hTERTp-null, Lv-hTERTp-tumstatin, Lv-hTERTp-TK, and Lv-hTERTp-tumstatin plus Lv-hTERTp-TK, respectively.Changes in tumor volume and weight, and cell morphology of tumor and major organs, were assessed by hematoxylin-eosin staining.Microvascular density of tumor tissue and cell apoptosis were assessed by immunohistochemical and TUNEL staining, respectively.

RESULTSThe Lv-infected HepG2 cells, and not the Lv-infected L02 cells, expressed TK and tumstatin. Lv-hTERTp-TK and Lv-hTERTp-tumstatin, alone or in combination, inhibited proliferation and increased apoptosis of the HepG2 cells, but the combination was more effective than either alone (P less than 0.05). None of the treatments affected proliferation or apoptosis of the L02 cells (P more than 0.05). The combination also led to a greater reduction of bcl-2 and VEGF than either alone (all, P less than 0.05). Tumor growth was significantly inhibited by the combination (P less than 0.05). In vivo, the combination treatment induced the greatest amount of apoptosis of the HepG2 cells. Cell morphology of major organs such as liver, spleen and kidney were similar to the control group. The combination also produced the most significant effect on tumor microvascular density (P less than 0.05) and the highest apoptosis index (P less than 0.05).

CONCLUSIONThe HTERT promoter can induce targeted expression of TK and tumstatin in HepG2 cells. Lv-hTERTp-TK combined with Lv-hTERTp-tumstatin can significantly inhibit proliferation and induce apoptosis of human HepG2 cells in vitro and in vivo, which may be related to down-regulation ofbcl-2 and VEGF.

Animals ; Apoptosis ; Autoantigens ; genetics ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Collagen Type IV ; genetics ; Down-Regulation ; Flow Cytometry ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lentivirus ; Liver Neoplasms ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Telomerase ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo study the effects of prenatal exposure to benzo[a]pyrene (B[a]P) on the physical development, early behavioral development, the adaptability to new environment and the learning and memory ability of rat offspring.

METHODSPregnant rats were randomly divided into five groups: control group, olive oil group, 3 exposure groups (25, 50 and 100 mg/kg B [a]P). The rats were exposed to B [a]P) by intraperitoneal injection on the 17th-19th days during gestation. The offspring were weighed on postnatal days (PND)1, PND 4, PND 7 and PND 28, the indices of physical development, reflective ability and sensory function were detected for offspring, the Morris water maze and Open-field tests were used to measure the ability of learning and memory and the adaptability to new environment of offspring.

RESULTSThe time of ear opening in middle and high-dose groups [(4.1 +/- 0.4),(5.0 +/-0.4) d] was posterior to that in untreated and solvent groups [(3.3 +/- 0.5), (3.4 +/- 0.6) d ](P < 0.01). The attainment rate (6.5%) of the surface righting reflex test in high-dose group on the 4th day was significantly lower than that (36.1%) in untreated group, the attainment rate (50.0%) in high-dose group on PND7 was significantly lower than those (81.3% and 79.3%) in untreated group and solvent group (P < 0.05). Compared to the untreated group, the time of forelimb hanging test in all exposure groups on PND12 and PND14 significantly decreased; compared to the solvent group the time of forelimb hanging test decreased in high-dose group on the 14th day significantly decreased (P < 0.01). The attainment rate (61.9%) of olfactory discrimination in high-dose group on PND12 was significantly lower than that (94.3%) in untreated group (P < 0.05). The results of Morris water maze test showed that the escape latency of different dose groups significantly increased, and the time of spatial probe and the times of traversing flat in high-dose group decreased significantly, as compared to the untreated and solvent groups (P < 0.01). The results of open-field test indicated that the center retention time in middle and high-dose groups significantly prolonged, the times of crossing lattice obviously reduced, and the rearing times decreased in high-dose group, as compared to untreated (P < 0.05).Compared to the solvent group, the times of crossing lattice in all exposure groups reduced significantly (P < 0.01 or P < 0.05).

CONCLUSIONThe prenatal exposure to B[a]P could inhibit the physical development and early behavioral development, and influence the adaptability to new environment and learning and memory ability for offspring.

Animals ; Benzo(a)pyrene ; toxicity ; Female ; Learning ; drug effects ; Maze Learning ; Memory ; drug effects ; Motor Activity ; Neurotoxicity Syndromes ; physiopathology ; Pregnancy ; Prenatal Exposure Delayed Effects ; physiopathology ; Rats ; Rats, Sprague-Dawley

Adult ; Aged ; Diabetes Mellitus, Type 2 ; etiology ; genetics ; Genetic Predisposition to Disease ; Humans ; Introns ; Middle Aged ; Obesity ; complications ; Polymorphism, Genetic ; Risk ; ras Proteins ; genetics

OBJECTIVETo investigate the optimal time and method of the early repair of the full layer eyelid defect caused by chemical burn.

METHODSFree nasal septum mucosal cartilage flap with muscle flap, skin grafting, or skin flap were performed in 18 cases (19 eyelids) with chemical burn within 4 postburn weeks. Eyelid reconstruction and corneal transplantation were performed at the same time in 4 patients.

RESULTSAll the reconstructed eyelids and transplanted cornea survived. The incidence of severe complications, such as exposure keratitis, corneal ulcer and eyeball perforation decreased.

CONCLUSIONFull layer eyelid defect caused by chemical burn should receive early reconstruction and repair, including timely reconstruction of eyelid for the sake of protecting the eyesight and of alleviating the inflammatory reactions, and the corneal transplantation should be done at the same time to avoid corneal perforation. Nasal septum mucosal cartilage flap could be ideal for the eyelid reconstruction.

Adult ; Burns, Chemical ; surgery ; Corneal Transplantation ; Eyelids ; injuries ; surgery ; Female ; Humans ; Male ; Middle Aged

Objective To construct a three-dimensional (3D) dynamic head-neck finite element model which ac cords with the anatomical structure,and study its dynamic responses under the extemal force.Methods By using the neck CT images of a Chinese adult male volunteer and obtaining the 3D cervical point cloud data,the finite element model of cervical spine was established using ICEM-CFD and HyperMesh software.This model,including vertebrae,intervertebral discs,facet joints,ligaments and cartilage tissues,and combining with the es tablished and verified head finite element model,was assembled as human head-neck finite element model with detailed anatomical structures.Results The model was validated by data of head-neck axial impact experiments reported in previously published literature.The simulation results showed that the neck deformation,head acceleration,head force and injury positions were preferably consistent with the experimental data.Conclusions The established 3D dynamic finite element model can be used to study head-neck dynamic responses and damage mechanism in the fields of traffic safety and impact injuries.

Reactive oxygen species generated by NADPH oxidase enhance aortic vascular smooth muscle cell proliferation and migration which play an important role in the pathophysiology of atherosclerosis. We investigated the role of NADPH oxidase in the cellular cholesterol metabolism in vascular smooth muscle cells using p47phox-deficient cells. Wild-type and p47phox knockout vascular smooth muscle cells were loaded with cholesterol for 72 h by using 10 mg/L cholesterol:methyl-beta-cyclodextrin complexes and then incubated with or without 0.3 mg/L thrombin for 10 min. Foam cell formation was determined by accumulation of intracellular cholesterol, oil Red O-stained lipid droplets. After cholesterol loading, cellular lipid droplets raised sharply, cellular cholesterol increased from (31.4+/-2.0) to (61.0+/-2.1) mg/g protein (P<0.05) in wild-type cells, and from (29.8+/-2.5) to (51.3+/-3.1) mg/g protein (P<0.05) in p47phox deficient cells, but the difference between the two cell types was not significant. Immunostaining showed decreased levels of smooth muscle alpha-actin and increased levels of macrophage marker Mac-2 in both wild-type and p47phox deficient vascular smooth muscle cells. One of the macrophage-related inflammation genes, monocyte chemoattractant protein-1 (MCP-1) expression did not change in both two cell types detected by immunostaining. Although additional incubating with thrombin, another macrophage-related inflammation gene, vascular cell adhesion molecule-1 (VCAM-1) expression was similar in all groups analyzed by real-time RT-PCR. However, the expression of ATP-binding cassette transporter A1 (ABCA1), acyl-coenzyme A:cholesterol acyltransferase 1 (ACAT1), the key proteins in cellular cholesterol metabolism, were similarly increased (P<0.05) in both two cell types as determined by quantitative real-time RT-PCR and Western blot, and it was not related to the state of oxidative stress. Interestingly, the expression of adipophilin, the lipid droplet related protein, had the similar results with ABCA1 and ACAT1, but, in wild-type cells, its expression also increased merely incubating with thrombin as determined by quantitative real-time RT-PCR. Together, these results suggest that p47phox-dependent NADPH oxidase is not involved in transdifferentitation of vascular smooth muscle cells into macrophage-like state after cholesterol loading. Deleting p47phox gene does not affect the cellular cholesterol metabolism in vascular smooth muscle cells.
ATP-Binding Cassette Transporters ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; metabolism ; Foam Cells ; cytology ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; enzymology ; NADPH Oxidases ; metabolism ; RNA, Messenger ; Sterol O-Acyltransferase ; metabolism ; beta-Cyclodextrins ; pharmacology

OBJECTIVETo investigate the analgesic and sedative effects of inhaling a mixture of nitrous oxide and oxygen on burn patient during and after dressing change.

METHODSA total of 240 burn patients hospitalized in the Institute of Burn Research of Changhai Hospital Affiliated to the Second Military Medical University, Department of Burns of the First People's Hospital in Zhengzhou, and Department of Burns and Plastic Surgery of General Hospital of Ningxia Medical University from October 2011 to September 2012 were enrolled in our study, and they were all in accordance with the inclusion criteria. The 240 patients were divided into control group (n = 60, treated with inhalation of oxygen during dressing change) and treatment group (n = 180, treated with inhalation of a mixture of 65% nitrous oxide and oxygen during dressing change) according to the computer-generated list of random number. The other treatments in control group and treatment group were the same. Before, during, and after dressing change, heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), oxygen saturation (SO2), and adverse effects were observed. The degree of pain and anxiety felt by the patients were respectively evaluated with the visual analogue scale (VAS) and Chinese version of the burn specific pain anxiety scale (C-BSPAS) at the same time points as above. Data were processed with analysis of covariance, chi-square test, analysis of variance, and rank sum test.

RESULTSThere were no significant differences between control group and treatment group in the levels of HR, SBP, DBP, and SO2 before dressing change (with F values respectively 0.76, 0.06, 1.11, 0.70, P values all above 0.05). Compared with those of control group, the levels of HR, SBP, DBP, and SO2 in treatment group were significantly ameliorated during dressing change (with F values respectively 81.78, 146.36, 226.44, 205.62, P values all below 0.01). After dressing change, the levels of DBP in the two groups were close (F = 0.31, P > 0.05), but the levels of HR, SBP, and SO2 showed statistical differences (with F values respectively 7.02, 8.69, 12.23, P < 0.05 or P < 0.01). Before dressing change, the VAS scores were approximate between control group and treatment group (Z = 0.21, P > 0.05). Compared with those in control group (9.4 ± 0.7, 1.7 ± 2.5), the VAS scores were significantly lowered in treatment group during and after dressing change (1.6 ± 1.3, 0.7 ± 1.1, with Z values respectively 11.84, 3.35, P values all below 0.01). There was no significant difference in C-BSPAS score between control group and treatment group before dressing change (Z = 0.62, P > 0.05). Compared with those in control group (75 ± 13, 73 ± 12), the C-BSPAS scores in treatment group were decreased during and after dressing change (9 ± 15, 9 ± 14, with Z values respectively 11.91, 12.28, P values all below 0.01). There were no obvious adverse effects in two groups before, during, and after dressing change.

CONCLUSIONSA mixture of nitrous oxide and oxygen seems to have obvious analgesic and sedative effects on burn patients during dressing change, and it can be widely used.

Administration, Inhalation ; Adolescent ; Adult ; Aged ; Analgesia ; methods ; Bandages ; Burns ; surgery ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Nitrous Oxide ; administration & dosage ; therapeutic use ; Oxygen ; administration & dosage ; therapeutic use ; Young Adult

OBJECTIVETo study the effect of Tiaomaiyin injection on the experimental arrhythmia for analyzing its underlying mechanism in the treatment of cardiovascular disease.

METHODExperimental animals anesthetized with 20% urethane (6 mL x kg(-1)) were evenly randomized into control group, positive control group, low-dose and high-dose Tiaomaiyin group. The rate of ventricular fibrillation (VF) chloroform-induced in mice, and the epoch of ventricular extrasystole (VE), ventricular tachycardia (VT),VF and cardiac arrest (CA), actonitine-induced in rats (1.0 microg x mL(-1) x min(-1)), and vabain-induced in guinea pigs (10 microg x mL(-1) x min(-1)), were detected respectively. The result loas converted into cumulative dosage of actonitine or vabain. In ischemia-reperfusion model in rats, the duration of arrhythmia and activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected.

RESULTAfter venous injection of Tiaomaiyin, VF in mice was lower significantly (P < 0.01), VE, VT, VF in rats and VF in guinea pigs were lowered considerably (P <0.05). The duration of arrhythmia in ischemia-reperfusion model was reduced considerably (P < 0.05), and the activity of myocardial SOD was raised significantly (P <0.01).

CONCLUSIONTiaomaiyin shows the reduction of experimental arrhythmia and protect effect to ischemia-reperfusion injury of the heart, which indicates that the effect mechanism may have the relationship with inhabition of lipid peroxidation and damnification of the free radical.

Animals ; Anti-Arrhythmia Agents ; administration & dosage ; isolation & purification ; pharmacology ; Arrhythmias, Cardiac ; etiology ; metabolism ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Electrocardiography ; Guinea Pigs ; Injections ; Malondialdehyde ; metabolism ; Mice ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; etiology ; metabolism ; physiopathology ; Myocardium ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the anatomical measurement of goat lumbar vertebrae and to compare with human lumbar vertebrae, so as to build the foundation for establishing animal models of lumbar prosthesis.

METHODSThe anatomical parameters of the vertebral body, pedicle and intervertebral disc in the fresh lumbar vertebrae of Boer goat and the lumbar vertebrae of healthy adults were collected by computer aided software Mimics16.0, and the anatomical characteristics of the two lumbar vertebrae were compared with the statistical software.

RESULTSThe anterior vertebral body height(VBHa) of goat lumbar was less than the middle vertebral body height(VBHm), which was less than the posterior vertebral body height(VBHp), and the maximum values were (38.7±2.9), (40.1±2.6) and (40.7±2.7) mm respectively. Its endplate width was greater than its depth, with the whole shaped like a heart or a kidney. The cranial endplate of goats was convex while the caudal endplate was depressed and the depression was small, with a maximum value of (1.6±0.6) mm. The pedicle height of goats increased from L₁1 to L̀ with the maximum of (30.5±1.9) mm; its pedicle width and angle increased firstly and then decreased with the increase of vertebra level and the minimum values were (6.7±0.4) mm and(45.9±2.6)° respectively. The anterior intervertebral disc height was larger than the middle which was larger than the posterior and all varied slightly with the changes of intervertebral spaces; the height and width of intervertebral foramen separately waved at (12.9±0.3) to (14.3±1.0) mm and (5.7±1.0) to (6.7±0.9) mm. The comparative results showed that the vertebral body height, pedicle height and angle of goats were greater than those of humans (<0.05) while the width and depth of the endplate, the intervertebral disc height, and etc. were significantly smaller than those of humans (<0.05). In addition, some structures, such as the height of pedicle and intervertebral disc, also showed different changing laws with the increase of vertebra level.

CONCLUSIONSAlthough there are similarities in goat lumbar spine in some aspects, such as endplate and foramen foramen, there are still many differences in many aspects. Understanding the anatomical characteristics of goat lumbar vertebrae and the difference between goat and human is of great guiding significance for the research of goat prosthesis and related technology.

Objective To investigate the rehabilitation needs and technical support for people with disabilities in rural areas. Methods From October, 2017 to February, 2018, 800 persons with disabilities, aged five to 80 years, from 23 villages in five township, Xi'an, Shaanxi, were surveyed with self-designed questionnaire and interview, including the basic situation, disability and training plan, rehabilitation support and skills maintained. Results The persons were mainly aged 50 to 80 years (58.37%), male (65.37%), accepting middle school education or less (46.63%), married (70.63%), living with their family (77.38%), income less than 2000 Yuan (66%), mainly from their family labor (62.62%). Their disabilities were mainly of grade 3 (40.63%), from hemiplegia (30.63%), dependence in living (45.38%), and no systematic rehabilitation program (55.25%). Most of them needed rehabilitation of self-care (40%), accessible guidance (30.63%) and reimbursement from medicare of Rural Cooperative Medical Scheme (50.63%), and hoped to participate social activities (41.87%). The rehabilitation supports were as that: the rehabilitation services were mobile or none (67.58%), the guiders of rehabilitation were few or none (48.88%), knew some or less rehabilitation knowledge (64.25%), the professionals accepted no continue education training (40%).Conclusion It is important to improve the rehabilitation technical support system for rural people with disabilities.