Objective To construct the eukaryotic expression vector containing short hairpin RNA (shRNA) targeting the inhibition of Smad3 gene segment in keloid fibroblasts (KFBs) and to investigate the effects on the expression of Smad7 in KFBs. Methods A couple of the most effective siRNA screened from former experiments were used to recombine the plasmids of Smad3 shRNA, and then Smad3 shRNA was transfected into KFBs by LyoVec TM. The expressions of Smad3 and Smad7 at different time points (0～9 d) were detected by RT-PCR and Western blotting. Results ① The recombinant Smad3 shRNA vectors were identified by RT-PCR and Western blotting. ② The expressions of mRNA and protein of Smad3 in KFBs decreased significantly in a time-dependent manner after transfection of Smad3 shRNA and reached the lowest point at 72 hours in the experimental group. Optical density analysis revealed a significant difference between the experimental group and the control group (P

CB-HRP was injected into the ovary of the domestic hens of 75-90 days old to trace the originating neurons of the vagus nerve innervated the ovary. The results were as the following:1. The afferent vagus neurons innervated the ovary were located in the nodose ganglia and the jugular ganglia. The afferent fibers in the ovarian medulla were found chiefly in the solitary tract, the nucleus of the solitary tract and the commissural nucleus of Cajal also.2. The efferent vagus neurons were located mainly in the subnueleus ventralis parvicellularis (VP)and the subnueleus ventrolateralis (VL)of the dorsal vagal motor nucleus, and a small number of neurons extended from the subnueleus VP and VL to the neighbouring five subnuclei.

BACKGROUND:Bone marrow mesenchymal stem cels have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cels is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cels can migrate to the thymus after transplantation.
OBJECTIVE:To observe the distribution and survival of bone marrow mesenchymal stem cels in the thymus of aging rats after transplantation.
METHODS: Bone marrow mesenchymal stem cels cultured in vitrowere transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cels were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cels homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation.
RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cels had a strong green fluorescence at days 3 and 7 after transplantation, and the cel contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P> 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cels were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cels can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.

OBJECTIVE:To investigate the cognition of clinical pharmacists on the clinical pharmacy practice and their profes-sion,and provide reference for further developing clinical pharmacy practice. METHODS:A questionnaire was designed,and ran-dom sample was conducted for the clinical pharmacists from 30 secondary and tertiary hospitals in Shanghai,and the investigation results were analyzed statistically. RESULTS:Totally 130 questionnaires were sent out,and 102 were effectively received with valid response rate of 78.46%. 94.11% of the investigated subjects were willing to be a clinical pharmacist,but 17.65% of them consid-ered that they were incompetent for their work;29.41% of them thought the current situation of clinical pharmacy practice was not ideal;all the respondents considered that the clinical pharmacist system should be implemented in medical and health institu-tions;the cognition and evaluation of investigated subjects in tertiary hospitals on clinical pharmacy practice and their profession was generally higher than those in secondary hospitals(P<0.05). CONCLUSIONS:The clinical pharmacists in medical and health institutions in Shanghai showed high cognition on their profession,and the clinical pharmacy practice has already made some achievements,but many work still need to be improved. Therefore,clinical pharmacists should continuously study their profession-al knowledge and improve their professional skills and quality;hospitals and universities should strengthen the subject construction and personnel training of clinical pharmacy and deeply carry out the clinical pharmacist system;health administration departments should strengthen the related laws and regulations of clinical pharmacy;colleges and universities should strengthen the discipline construction and personnel training.

Objective To investigate the effects of propofol on ketamine-induced HSP 70 mRNA and protein expression in the rat posterior cingulate cortex and to explore the possibility of using propofol to prevent or treat ketamine-induced psychotomimetic effects and neuronal damage. Methods Thirty male Wistar rats weighing 250-300 g were randomly divided into 5 groups with 6 animals in each group: group 1 received normal saline intraperitoneally ip (NS); group 2 received ketamine 100 mg? kg-1 ip (K); group 3 received propofol 100 mg ? kg-1 ip (P); group 4 received propofol 50 mg?kg-1 + ketamine 100 mg?kg-1 ip (P1 K) and group 5 received propofol 100 mg?kg + ketamine 100 mg?kg-1 ip (P2K) . In group 4 and 5 the interval between propofol and ketamine administration was 15 min. Twenty-four hours after ketamine and/or propofol administration, the animals were decapitated and brain was removed. HSP 70 mRNA expression in the posterior cingulate cortex was detected by using semi-quantitative RT-PCR technique; HSP 70 protein expression in posterior cingulated cortex was determined by immuno-histochemical method. Results The levels of HSP 70 mRNA and HSP 70 protein expression were significantly different among the 5 groups. Ketamine induced marked HSP 70 mRNA and HSP 70 protein expression in the posterior cingulated cortex. Propofol itself did not induce HSP 70 gene expression in this brain region. Propofol significantly inhibited ketamine-induced HSP 70 mRNA and HSP 70 protein expression in the posterior cingulate cortex in a dose-dependent manner. Conclusions Propofol pretreatment can significantly inhibit ketamine-induced HSP 70 mRNA and protein expression in the posterior cingulated cortex. It may be one of the mechanisms of inhibition of ketamine-induced psychotomimetic effect and neuronal damage by propofol.

Objective:To investigate the regulatory mechanism of LBP on T lymphocytes excessive apoptosis and the mRNA expression of apoptotic genes in aged rats Methods:T lymphocyte apoptosis and the expression of pre apoptotic and anti apoptotic genes (Fas, FasL, TNFR1, Bax, Bcl2 and TNFR2) from young rats and old rats treated with or without LBP were studied using flow cytometry and fluorescence real time quantitative RT PCR Results:LBP could effectively downregulate the percentage of T cell apoptosis in aged rats At the same time LBP also downregulated the transcription level of TNFR1 gene and upregulated the transcription level of Bcl 2 in T cells from old rats Conclusion:The T cell excessive apoptosis in old rats could be effectively downregulated with LBP, by the by of decreasing the expression of pro apoptotic gene and enhancing the expression of anti apoptotic gene

To treat large facial defect (more than 6 cm x 4 cm in diameter) or a wound with bone exposure to atmosphere by less-traumatic, easier-healing reconstruction method, a pedicle flap including facial, neck, posterior auricle and occipital skin flap was designed and transferred, one by one, to repair facial defect as well as other flap donor sites, but occipital skin flap was only used to cover posterior auricle area. After 2-3 years follow-up, well-healed skin flaps with good color, elasticity and sensation were observed in all 16 patients. It is concluded that this method is effective and practical.
Carcinoma, Basal Cell/surgery ; Cicatrix/*surgery ; Face ; Neck ; Orbital Neoplasms/*surgery ; Parotid Neoplasms/*surgery ; Reconstructive Surgical Procedures/methods ; Scalp ; *Surgical Flaps

BACKGROUND:Studies have shown that exogenous bone marrow mesenchymal stem cells can settle down in lung tissue, participate in long regeneration, but few studies concerned the repair of aging lung injury. OBJECTIVE:To observe the effect of bone marrow mesenchymal stem cells on lung injury induced by D-galactose. METHODS:A total of 30 Sprague-Dawley rats were equal y divided into three groups at random:control group, aging model group and celltreatment group. To establish the aging rats, 10 rats each in the aging model group and celltreatment group were daily subcutaneously injected with D-galactose for 4 months. 3×106 bone marrow mesenchymal stem cells were transplanted via caudal vein in the celltreatment group, once a week, for 4 weeks. cellmedium of equal dose was added in the control and aging model groups. Bone marrow mesenchymal stem cells were transfected by lentiviral vectors expressing green fluorescent protein to determine the implantation of bone marrow mesenchymal stem cells in rat lung. Superoxide dismutase activity and malondialdehyde content in rat lung were measured in each group. The difference in rat lung structure was observed using hematoxylin-eosin staining in each group. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells marked by green fluorescent protein were implanted in rats, migrated towards lung tissue and survived. Compared with aging model group, superoxide dismutase activity was apparently increased, but malondialdehyde content was obviously diminished in the celltreatment group. In each group, histopathological sections revealed that normal pulmonary alveolus was damaged in the aging model group, showing enlarged air cavity and emphysema. Lung injury was evidently repaired inthe celltreatment group. Results suggested that bone marrow mesenchymal stem cells could repair lung injury in aging rats, and exert anti-aging effects.

ObjectiveTo compare the accelerating effect of topical recombinant human epidermal growth factor (rhEGF) and recombinant bovine basic fibroblast growth factor (rb-bFGF) on wound healing after fractional CO2 laser therapy.MethodsTwenty male guinea pigs were included in this study.After hair removal and irradiation with fractional CO2 laser,the back of each guinea pig was divided into 4 regions to be topically treated with rhEGF of 10 μg/cm2 (rhEGF group),rb-bFGF of 262.51 IU/cm2 (rb-bFGF group),the combination of rhEGF and rb-bFGF (combination group),or normal saline (control group),twice daily until the healing of wound.Skin physiology parameters including elasticity index and melanin index were detected before the irradiation,7,14 and 28 days after the irradiation,and compared between the 4 groups by analysis of variance.Tissue specimens were obtained from 4 mice at the above time points and subjected to pathological examination for the observation of collagen fibers and quantification of fibroblasts.ResultsAfter fractional CO2 laser therapy,the crusts fall off completely in growth factor-treated regions,while partly in the control regions,within 3 to 7 days; the wounds healed completely in 14 to 28 days in all the groups,with the regenerating tissue being more tender and redder compared with the surrounding unirradiated tissue.The wound surface was smaller in area and redder in color in the 3 growth factor-treated groups than in the control group.At 28 days after the irradiation,the elasticity index was 262.29 ± 62.40 in the combination group,202.00 ± 65.62 in the rhEGF group,188.86 ± 35.02 in the rb-bFGF group,167.14 ± 42.49 in the control group.Statistical difference was observed in elasticity index,but not in skin melanin index among the 4 groups.Pathological examination showed a dense and organized arrangement of collagen fibers in the combination group but a sparse and disorganized arrangement of collagen fibers in the control group.ConclusionThe combined application of rhEGF and rbbFGF can accelerate the healing of wound and increase the elasticity of regenerating tissue after fractional CO2 laser therapy.