BACKGROUND:Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cel s (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cel transplantation. OBJECTIVE:To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS:BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cel s under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Al en’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the model group were given no treatment. Five weeks after modeling, spinal cord samples were taken from each rat to make frozen sections for detection of nerve related markers RESULTS AND CONCLUSION:After 48-hour adenoviral transfection, Shootin1 expression was successful y detected in BMSCs. After 7-day in vitro induction, the cel morphology in the three groups varied, and there was no significant difference in the expression of Nestin and NeuN between the transfection and non-transfection groups. Basso, Beattie and Bresnahan scores were higher in the two cel transplantation groups than the model group. Increased expression levels of Nestin, NeuN, GFAP, MAP-2, ChAT and SYN were observed in both two cel transplantation groups, indicating a strengthened ability of nerve regeneration. Our experimental findings further confirm that BMSCs transplantation for spinal cord injury has achieved good outcomes, and Shootin1 protein plays a certain role in nerve regeneration and functional recovery after spinal cord injury. However, Shootin1 overexpression shows no obvious additional effects in combination with BMSCs transplantation, and further studies on the optimization of BMSCs transplantation for spinal cord injury are necessary.

Objective To investigate the correlation between clinicopathological parameters and the expressions of inhibitor of differentiation/DNA binding 3 (Id3), vascular endothelial growth factor (VEGF) and human epidermal growth factor receptor-2 (HER-2/C-erbB-2) in patients with breast cancer. Methods The protein expressions of Id3, VEGF and C-erbB-2 were detected by SABC immunohistochemistry in 70 spceimens of breast cancer, 40 specimens of tissue adjacent to breast cancer, 20 specimens of fibroadenoma and 20 specimens of normal breast tissue. Results The expressions of Id3, VEGF and C-erbB-2 in breast cancer tissue (88.6%, 95.7% and 20.0%, respectively) were significantly higher than that in the other tissues (P0.05), while the expression of C-erbB-2 protein substantially varied among the patients of breast cancer in different pTNM stages (P

ObjectiveTo study the feasibility of laparoscopic assisted splenectomy for massive splenomegaly and devascularization in the treatment of patients with cirrhosis and portal hypertension.MethodA retrospective study was carried out on 12 patients who underwent laparoscopic assisted splenectomy and devascularization from October 2003 to January 2010.ResultsThe operation was successfully carried out in eight patients.The remaining four patients were converted to open surgery due to intra-operative bleeding.The mean operation times for the 12 patients and the 8 patients who underwent laparoscopic surgery were 350 min (range,190-470 min) and 313 min (range,190-410 min),respectively.The corresponding mean intra-operative blood losses were 1300 ml (range,200-2200 ml) and 700 ml (range,200-1700 ml),respectively.For the 8 patients who received laparoscopic surgery,flatus were passed 3-5 days after operation,and the postoperative hospitalization was 10-20 days.ConclusionsLaparoscopic assisted splenectomy for massive splenomegaly and devascularization is feasible.It is a treatment option for patients with cirrhosis and portal hypertension.

BACKGROUND: Most of the literatures only reported that there is a great difference between diabetics who have a high 24-hour urinary albumin excretion rate and those without renal damage, but there is no obvious difference between cases of microalbuminuria and those without renal damage.OBJECTIVE: To probe into the relationship between diabetic nephropathy and osteoporosis.DESIGN: A case-control study.SETTING: Department of Endocrinology, West China Hospital of Sichuan University.PARTICIPANTS: According to the diagnostic standard set by the American Diabetes Association in 1997 (fasting blood glucose ≥ 7 mmol/L,postprandial blood glucose ≥ 11.1 mmol/L), 96 diabetic patients were selected, including 56 males ＜ 60 years old and 40 females who had not entered the menopausal period, excluding the influence of age and osteoporosis caused by menopause. The average age of the subjects was (48.7±10.5)years, their duration were from one month to twenty-one years with the aver age of (7.85±2.56) years, and their general information had no significant differences.METHODS: According to the urine albumine excretion rate and renal function, the patients were divided into four groups: normal albuminuria group (n=48), microalbuminuria group (n=28), macroalbuminuria group (n=15), renal failure group (n=5). The bone mineral densities of lumbar spines (L2-4), femoral neck, Ward's triangle and trochanter were detected with Dual energy X-ray absorptiometry, and then the fasting blood glucose,2-hour postprandial blood glucose, glycosylated hemoglobin A, alkaline phosphatase, calcium, phosphorus, blood urea nitrogen, creatinine and urine albumine excretion rate were compared between the patients with and without osteoporosis.MAIN OUTCOME MEASURES: The fasting blood glucose, 2-hour postprandial blood glucose, glycosylated hemoglobin A, alkaline phosphatase, calcium, phosphorus, blood urea nitrogen, creatinine and urine albumine excretion rate, as well as the bone mineral density, were observed in all the patients.RESULTS: The L2-3 bone mineral density in the macroalbuminuria group was significantly different from that in the microalbuminuria group (P ＜ 0.05). The proximal femur bone mineral density in the microalbuminuria group was significantly different from that in the normal albuminuria group (P ＜ 0.05). The bone mineral densities of proximal femur and lumbar spine in the renal failure group were significantly decreased as compared with those in the other groups (P ＜ 0.01). The disease course,glycosylated hemoglobin A, alkaline phosphatase and body mass index were significantly different between the patients with and without osteoporosis (P ＜ 0.05).CONCLUSION: The incidence rate of osteoporosis is increased with the aggravation of nephropathy, and diabetic nephropathy may be closely correlated with the decrease of bone mineral density and the occurrence of osteoporosis.

Objective To clarify the effect of antiproliferative factor（APF）on the proliferation of bladder cancer cell line T-24.Methods Active or no active APF with different concentrations was added to cultured bladder cancer T-24 cells.Methyl thiazolyl tetrazolium（MTT） assay was used to determine the proliferation of T-24 cells.The expression of c-Jun and HB-EGF were examined by Western blot.Results MTT showed that there was significant difference in the proliferation with the concentration of AFP increased from 0.5 μg /ml to 1.0 μg /ml.Compared with the negative control group,the absorption value of no active APF group showed no change（P 〉 0.05）.Western blot indicated that the c-Jun and HB-EGF expression were down-regulated with the concentration of active APF increased（P 〈 0.05）.No difference in their expression was found in no active APF groups.Conclusion Active APF may decrease the expression of c-Jun and HB-EGF,so as to inhibit the proliferation of bladder cancer cells.Our experiment might provide evidence for the therapeutic applications of APF for the bladder cancer.

Objective To study the time distribution of recurrence for postoperative breast cancer patients by hormone receptor status.Methods The characteristics of recurrence in 1099 breast cancer patients with different hormone receptor status undergoing surgery between December 1999 and April 2006 were analyzed retrospectively.Results For those 1099 patients the median follow-up time was 60.6 months.Recurrence was found in 171 patients.For hormone receptor-negative (HRN) patients the first peak of recurrence appeared at the 12th month and the second at about the 54th month.For hormone receptor-positive (HRP) patients the peak of recurrence appeared at the 36th month and a second peak at about the 54th month,and beyond 54 months the hazard was higher for hormone receptor-positive patients.The risk of recurrence was higher with more nodes involved.Node-positive HRP patients suffered two to three times higher risk of recurrence than node-negative HRP patients.Node-positive HRN patients had three to four times higher risk of recurrence than node-negative HRN patients.The recurrence-free survival in HRP patients was higher than that in HRN patients,also the recurrence-free survival in node-negative HRP patients was higher than that in node-positive patients (all P ＜ 0.01).Conclusions The recurrence risk for HRP breast cancer patients was higher than that in HRN patients after 54 months postoperatively.The risk of recurrence for node-positive breast cancer patients was higher than that for HRP node-negatives.

Objective To study the effects of 1,25-(OH)_2-vitamin D_3[1,25-(OH)_2D_3] on proliferation,differentiation, and secretion of osteoprotegerin and RANK ligand (RANKL)in cultured marrow mesenchymal cells of rhesus monkey (RhBMSCs). Methods Three different concentrations of 1,25-(OH)_2D_3 (10~(-12) ,10~(-10), and 10~(-8)mol/L)were added to the cultured RhBMSCs in vitro. MTT was used to observe cell proliferation and ELISA technique was used to measure the concentration of alkaline phosphatase (ALP), osteocalcin, osteoprotegerin, and RANKL. Mineralization nodus was identified via alizarin red staining. Results Under different concentration of 1,25-(OH)_2D_3, RhBMSCs proliferation were promoted within 7 days but were suppressed beyond 7 days. No significant dose-dependent manner was found. Differentiation of RhBMSCs into osteoblast was promoted by 1,25-(OH)_2D_3. The levels of ALP and osteocalcin in groups with various concentrations of 1,25-(OH)_2D_3 were higher than those of control group [ALP(ng/ml) ,7 d:31.40±1.25,26.50±0.50,28.47± 0.25 vs 13.48±0.26;10 d:33.37±0.68,35.30±1.57,33.27±0.67 vs 17.14±0.55;13 d:35.37±0.12,30.47± 0. 25 , 30. 27±1.25 vs 16.55 ± 1.13 ; osteocalcin (ng/ml), 7 d:4.47±0. 29,4.00 ±0. 60,3.73±0.78 vs 1.63± 0.55;10 d:5.63±0.57,5.17±0.15,4.30±0. 10 vs 2.17±0. 15;13 d:7.03±0.15,5.53±0.25,5.27±0.31 vs 2.23±0. 55 ; all P < 0. 05], but no typical mineralization nodus was found in either group. Secretions of osteoprotegerin and RANKL from RhBMSCs were stimulated by 1,25-(OH)_2D_3 [osteoprotegerin (pg/ml), 7 d: 72.57±0.67,68.00±1.75,64.23±0.87 vs 30. 13±1.72; 10 d:62.03±1.62,51.80±1.30,28.93±0.95 vs 18.13±1.40;13 d:65.13±0.71,62.43±2.11,44.93+1.63 vs 36.70±0.95 ;RANKL(pg/ml) ,7 d:74.33+0.61, 82.37±2.15,85.23±0.45 vs 70.83±1.71 ;10 d:83.30±0.46,86.70±0.56,88.23±0.91 vs 74.20±1.83;13 d: 81.70±1.81,81.07±0.95,84.70±1.41 vs 72.73±0.97 ;all P<0.05]. The secretion of RANKL was increased at first and then decreased, whereas the secretion of osteoprotegerin had the opposite tendency. The secretions of RANKL and osteoprotegerin were both in a dose-dependent manner. Conclusions 1,25-(OH)_2D_3 promotes differentiation of RhBMSCs into osteoblasts,resulting in increased secretions of osteoprotegerin and RANKL

Objective In this research ,the effects of exogenous C 2-ceramide on the induction of apop-tosis of mesothelioma cells in vitro and related important proteins are investigated .Mtehods Mesothelioma cells were treated with various doses of C 2-ceramide for different duration .Cell viability were analyzed by cell count-ing kit-8 assay.Morphological changes of apoptosis of mesothelioma cells were observed by Diff Quik staining . Apoptosis of mesothelioma cells was also detected by Caspase -3 assay and lactate dehydrogenase(LDH)assay. Related important proteins involved in the signal transduction of apoptosis were detected by western blot .Results In vitro,C2-ceramide demonstrated a dose -and time-dependent inhibition of cell proliferation .Apoptotic bodies were observed by Diff Quik staining .The antiproliferative effect of 80 μm C2-ceramide was paralleled with an increase in Caspase -3 activity.LDH assay showed that C2-ceramide at a concentration of 80μm signifi-cantly promoted cells death .After treated by C2-ceramide,the expressions of Bax and the phosphorylated JNK in mesothelioma cells were increased , however , the expression of phosphorylated ERK 1/2 kinase was decreased . Conclusion Our results indicate that C 2-ceramide induced apoptosis of malignant mesothelioma cells in vitro . This anti-tumor affect is achieved by adjusting related important proteins .

Objective To observe the local field potential activity of dorsomedial and dorsolater striatal (STR) neurons in rats with exercise-induced fatigue,so as to explore the regional characteristics of those neurons.Methods Thirty-six male Wistar rats were randomly divided into a control group(CG),a single fatigue group(SFG)and a repeated fatigue group(RFG),each of 12.The exercise-induced fatigue model was established according to a load-increasing treadmill running protocol.The CG did not do any treadmill running,SFG finished one-time exhaustive exercise while the RFG repeated exhaustive exercise 6 times.The extracellular glass microelectrode technique was used to record the spontaneous firing of dorsomedial(n=6)and dorsolateral(n=6)STR neurons in vivo to observe the discharging frequency,characteristics and types.Results (1)Spontaneous firing frequencies of striatal neurons in SFG were significantly higher than those of CG(P＜0.01),while those of RFG were significantly lower than SFG.(2) After repeated fatigues,there were significant decreases in the irregular firing pattern and significant increase in the explosive firing of striatal neurons compared with CG(P＜0.05).(3)After the single fatigue and repeated fatigue,the discharging frequency of dorsomedial medium spiny-like neurons increased significantly(P＜0.05),and that after repeated fatigue significantly higher than that of the dorsolateral(P＜0.01).The discharge frequency of the dorsomedial and dorsolateral fast-spiking neurons decreased significantly after repeated fatigues.The discharge frequency of dorsolateral large aspiny-like neurons of RFG was significantly higher than SFG,while that of the latter was significantly lower than CG(P＜0.05).Conclusions (1)Striatal neurons mediate exercise-induced fatigue and present regional characteristics,which might be due to dorsomedial and dorsolateral striatal neurons receiving different types of projection neurons.(2) Striatal fast-spiking neurons may play an important role in mediating the exercise-induced fatigue.

Objective To discover the effects of the Prescription Administrative Policy in force on the quality of the prescriptions in a tertiary hospital in 2007. Methods The prescriptions of 400 cases were sampled systematically for evaluation, and 60 patients were interviewed. Results The average eligibility rate of the prescriptions was but 37. 2% in this hospital, which was mainly plagued by incompleteness, nonstandard and irrationality found in prescriptions. Implementation of the Policy contributed to a significant improvement of some indicators. For example, the eligibility rate increased by 12. 2% (P=0. 004) ,the percentage of the use of antimicrobial agents dropped significantly (P=0. 001),and the percentage of generic names used rose significantly (P = 0. 000). Conclusions The implementation of the Policy is highly positive for prescription quality.