Objective pSUPER was used as vector to construct the siRNA targeting the gene of Rac1 to explore its inhibitive effects on Rac1 expression in hepatoma cell HepG2,and observe the proliferation of HepG2.Methods The oligonucleotide possessing 64 bases targeting the hairpin-like RNA of Rac1 gene was chemically synthesized,and inserted into the downstream of H1 promoter of linearized pSUPER vector after annealing.The vector pSUPER-Rac1-siRNA was constructed by sequencing and restrictive digestion of the recombined vector,and then transfected to hepatome cell line HepG2.pSUPER-Control-siRNA was used as a negative contro1.Western blotting was performed to monitor the Rac1 gene in plasmid-transfected cells,and the proliferation of extracorporeal cells after Rac1 expression was detected by MTT assay.Results The siRNA expression vector pSUPER-Rac1-siRNA targeting Rac1 gene was successfully constructed.The expression of Rac1 protein in HepG2 cells was down-regulated effectively by the transfection of pSUPER-Rac1-siRNA with an inhibition rate of 82%.It was revealed by MTT detection that the proliferation rate of cells declined remarkably when the expression of Rac1 protein was inhibited efficiently.Conclusions The hairpin-like RNA expression vector pSUPER-Rac1-siRNA targeting Rac1 has been constructed successfully,which may effectually and specifically down-regulate the gene expression of Rac1 in HepG2 cells.The silencing of Rac1 protein may remarkably inhibit the proliferation of hepatoma cells,implying that the Rac1 may play an important role in the in vitro growth of hepatoma cells.

OBJECTIVE:TO summarize the treatment of complicated proximal humeral fracture with artificial shoulder replacement.METHODS:A computer-based online search of VIP was performed for related articles published between January 1998 and October 2009 with the key words"shoulder joint,prosthesis replacement,proximal humeral fracture".Articles with high correlation were included.and repetitive studies were excluded.The data were firstly collected,the references of each article were reviewed.RESULTS:A total of 18 articles were included The treatment of complicated proximal humeral fracture remains controversial.Early semi-shoulder ioint replacement is an appropriate method.Shoulder replacement includes humeral head replacement and total shoulder replacement.Neer and Biligani type are typical artificial humeral head.The fixation patterns of humeral head prosthesis shaft include cemented and cementless.Bone cement includes total bone cement and proximal cement fixation,and cementless includes pressure fixation and compaction bone grafting.Complications following shoulder replacement involve prosthesis instability,nodule heterotopia,heterotopic ossification,prosthesis loosening,periprosthetic fracture,infection and nerve injury.CONCLUSION:Majority of complications can be prevented by appropriate location of prosthesis,reconstruction of greater and lesser tubercle.and in combination with postoperative rehabilitation treatment.

BACKGROUND:Mouse nerve growth factor can promote the repair and regeneration of injured nerves, but current experimental research shows that the effects of different treatment methods are stil controversial.
OBJECTIVE:To evaluate the effect of mouse nerve growth factor injection via different ways on the treatment of peripheral nerve injury.
METHODS:Total y 52 patients with peripheral nerve injury were randomly assigned into two groups:experimental group (local injection of mouse nerve growth factor, n=27) and control group (systemic administration of mouse nerve growth factor, n=25). The treatment was performed once a day, and lasted for 4 weeks. Then, the clinical efficacy and recovery of neurological function were compared.
RESULTS AND CONCLUSION:The good and effective rates were 85%(n=23) and 93%(n=25) in the experimental group, while 72%(n=18) and 84%(n=21) in the control group, respectively, which were significantly better in the experimental group than the control group (P<0.05). In the experimental group, 13 cases developed transient pain at injection site, including one case of remission undergoing oral analgesics;in the control group, 12 cases had transient pain at injection site, without any treatment. The results suggest that both local and total body injection of mouse nerve growth factor are safe and effective for treatment of peripheral nerve injury, but local injection is superior to systemic administration.

【Objective】To observe the effect of plasma ?-endogenous peptides (?-EP) on gestational function in endometriosis(EMT) rats.【Methods】 Female SD rats at the third estrous cycle were used to establish EMT model.Then 20 EMT rats were equally randomized into the model group and naltrexone(an opioid receptor antagonist) group,and other 10 rats were in the pseudo-operation group.Naltrexone group was given naltrexone 2?mg?kg~-1?d~-1 by gastric gavage during proestrus and normal saline at other time,and the model group and pseudo-operation group were given normal saline for one month.Serum levels of ?-EP,luteinizing hormone(LH) and prolactin(PRL) were examined during proestrus,and serum progesterone(P) level and gestational function were observed after estrus.【Results】Serum levels of ?-EP and PRL increased and LH decreased during proestrus,and serum P level also decreased after estrus in the model group(P

Objective To observe the effects of nAChR antagonistα-conotoxin Eb1.6 on ther-mal pain threshold and spinal IL-1βexpression levels and astrocytes activation in rats using L5 spinal nerve transaction (SNT)model.Methods Fifty male Sprague-Dawley rats were randomly assigned into 5 groups with each group 10 rats:sham group,different doses of α-CTX Eb1.6 (0.1 5,1.5 and 1 5 nmol/kg)groups and the saline group after SNT.Saline solution or different doses of Eb1.6 were intraperitoneally injected seven days after the surgery when the model was stable and the treatment continued for seven days.Measured the TWLs of all groups of the rats 1,2,4,7,12 hours after the in-jection on 7 d and 13 d.The rats were sacrificed and L5 spinal cord tissues were collected immediately after the behavioral tests on 13 d.The expression of GFAP and IL-1βwere assessed by Western blot assay and enzyme linked immunosorbent assay (ELISA)separately.Results Groups E1,E2,E3 and C had shorter TWL before the injection on 7 d and 13 d than group N(P <0.05).The TWLs of the rats in groups E1,E2 and E3 of 1 h,2 h and 4 h after the injection on 7 d were significantly higher than that before the injection(P <0.05)with 2 h after the injection showed the most obvious change.The TWL of 1 h,2 h,4 h and 7 h after the injection of the rats in group E1,E2 and E3 and those of 12 h after the injection of the rats in group E2 and E3 on 13 d were significantly higher than that before the injection(P <0.05 )and also higher than TWL of the respective time points on 7 d(P < 0.05 ),also with 2 h after the injection showed the most obvious change.The TWLs of 2 h after the injection a-mong group E1,E2 and E3 showed significant differences both on 7 d and 13 d (P <0.05).Rats spi-nal IL-1βand GFAP expression levels of group E1,E2,E3 and C were significantly higher than those of group N(P <0.05).Rats spinal IL-1β and GFAP expression levels of groups E1,E2,E3 signifi-cantly decreased compared with group C(P <0.05).There were significant differences among the spi-nal IL-1βand GFAP expression levels of group E1,E2 and E3(P <0.05).Conclusion Eb1.6 dose-de-pendently reduced the thermal hyperalgesia induced by L5 spinal nerve transection.Repeated treat-ment of Eb1.6 could produce better analgesic effect,which might be partly attribute to the inhibition of spinal IL-βlevels and astrocytes activation.

Cervical anastomosis by the thoracic approach for the treatment of upper esophageal cancer can simplify surgical steps and reduce incidence of anastomotic leak. This approach has been used for 26 patients with upper esophageal cancer who were admitted to the Jiangsu Cancer Hospital from July 2006 to August 2009. The mean length between lesion and incisor was 23.3 cm. General anesthesia and double-lumen intubation through left posterolateral incision in the fifth intercostal space was adopted. The stomach was dissociated with the technique of "in situ dissociation", and esophagus was dissociated conventionally. Double purse-string suture was adopted to fix the esophageal mucosa onto the supportive base of the stapler, and make purse-string suture to fix stomach on the center pole of the stapler. There was one failure case which has been converted to the manual cervical anastomosis, and the operations for the rest 25 cases were completed successfully, without anastomotic leakage and positive margin. The average blood loss was (352 ±211 )ml, and the average operation time was (3.7 ±0.6 )hours.

Objective To research the mechanism of the changes of T lymphocyte subtypes and provide reference for clinically prevention,diagnosis and treatment for NSCLC through analysis of the expression of Th1 ,Th2 in Non-small-cell carcinoma (NSCLC)patients.Methods Whole blood (EDTA anticoagulant treatment)from 60 NSCLC patients and 60 healthy sub-j ects were collected to detect of the expression of CD3 +T cells,CD4+T cells and CD8+T cells on T lymphocytes and the lev-els of Th1 and Th2 cells by flow cytometer (FCM),and the absolute value of T lymphocyte by hematology analyzer.Results Compared with normal control group,after surgery 1~3 days NSCLC groups,the percent of CD3 +T,CD4+T,CD8+T cells and the CD4+/CD8+ ratio in the NSCLC patients before surgery were significantly reduced 58.40±10.27 vs 66.58± 6.84,31.32±8.65 vs 39.40±6.43,34.23±8.00 vs 24.31±8.16,0.96±0.23 vs 1.58±0.23 (t=-6.726~14.916,P<0.05).The percent of CD3 +T,CD4+T,CD8+T cells and the CD4+/CD8+ ratio in the NSCLC patients after surgery 1~3 days were also significantly decreased 56.31±8.00 vs 66.58±6.84,27.72±7.55 vs 39.40±6.43,33.69±7.10 vs 24.31± 8.16,0.87±0.31 vs 1.58±0.23 (t=-6.720~14.367,P<0.05).The percent of CD4+T cells in the NSCLC patients af-ter surgery 4~7 days was increased 33.23±4.13 vs 39.40±6.43(t=6.257,P<0.05).Compared with the control group, within the helper T cell subsets,the cell content of Th1,Th2 cells (× 10/μl)and the Th1/Th2 ratio were significantly changed in different extent in the NSCLC group before surgery 6.79±1.34 vs 12.52±3.56,4.82±0.51 vs 2.32±0.82, 1.39±0.84 vs 5.36±1.42 (t=-20.087~18.630,P<0.05).The content of Th1 cells was lower in the NSCLC patients after 1~3 days and 4~7 days 8.86±1.52 vs 12.52±3.56,7.02±1.27 vs 12.52±3.56 (t=7.339~11.275,P<0.05). Conclusion The NSCLC patients presented immune dysfunction,like T lymphocytes and helper T cells decreased and Th2 cells were clearly in the ascendant.Also,the cytotoxic T cells increased by the stimulation of cancer cells,but they began to decrease after the surgery.

Objective To explore the effects of intrathecal injection of acetyltransferase p300 inhibitor Garcinolon on hyperalgesia in a rat model of L5 spinal nerve ligation and its underlying mechanism. Methods After lumber intrathecal catheters implanted,90 male Sprague-Dawley rats (40-50 d,weighing 180-220 g)were randomly divided into six groups (n=15 each):Naive group (group N),Sham operation group (group S),SNL group (group C)and three Garcinol treatment groups (group H:500μg/kg,group M:100μg/kg,group L:20μg/kg).Group N did not receive any operation in rats.In group S,the L5 spinal nerve was only exposed without ligation.Other four groups all received spinal nerve ligation (SNL).Group N,group S and group SNL were administrated intrathcelly with 100% DMSO (10 μl)within 3 to 6 days after SNL surgery.Other three groups were treated with Garcinol 500μg/kg (group H),100μg/kg (group M),20 μg/kg (group L)at the same points respectively.Behaviorally,thermal withdrawal latency (TWL) were measured at 1 day (T0 )before and on days 1(T1 ),3(T2 ),5(T3 ),7(T4 ),9(T5 ),11(T6 ),14(T7 )after surgery separately.On the 7th day after behavioral testing,the lumber segment of the spinal cord was removed to test the level of p300 and acetyl-p65 by western blot,while NF-κB was detected with immunofluorescence. Results Compared with group N,TWL was significantly shortened in group C,L,M and H,the levels of ace-tyl-p65 and p300 in group C,L,M and H were markedly increased,the expression of NF-κB in group C,L,M and H was markedly increased(P<0.05).Compared with group C,TWL was significantly prolonged in group M and group H,the levels of acetyl-p65 and p300 in group M and group H were dramatically decreased,the expression of NF-κB in group H was obviously decreased(P <0.05).Conclusion The acetyltransferase p300 inhibitor Garcinol imposes protective effect in SNL-induced neuropathic pain.Mechanisms are probably associ-ated with decreasing acetyl-p65 protein expression level in the NF-κB pathway.

Aim To investigate the antagonistic effect of resveratrol on neuropathic pain and its underlying mechanism. Methods Neuropathic pain was induced by ligation of L5 spinal nerve (SNL) in rats. 90 male Sprague-Dawley rats, fit with intrathecal catheters were divided randomly into six groups ( n = 15 ): naive group; sham group; SNL group; high dosage of res-veratrol group (300μg);middle dosage of resveratrol group ( 30μg ) and low dosage of resveratrol group (3μg). The naive group did not make any process. In sham group, the L5 spinal nerve was only exposed without ligation. Other groups received SNL. Different dosages of resveratrol dissolved in 10μL 100% DMSO were administered by intrathecal injections once a day for 4 days, starting on day 4 after SNL. Paw withdraw-al latency (PWL) was measured on day 1,3,5,7,9, 11,14 days after surgery separately. On day 7 after be-havioral testing, the lumbar segments of the spinal cord were removed to measure the level of SIRT1 and acety-lated-p65(Ac-p65) for western blot. The activation of NF-κB was determined through calculating the percent-age of NF-κB-immunofluorescence positive staining cells in this study. Results Compared with sham groups,the SNL group showed an obvious decrease(P< 0. 05) of PWL and SIRT1 after surgery,whereas Ac-p65 and actived NF-κB significantly increased ( P <0. 05) in the spinal cord. Administration with high and middle dosages of resveratrol markedly attenuated(P <0. 05) SNL-induced thermal hyperalgesia and down-regulation of SIRT1 and blocked (P < 0. 05) the SNL-induced up-regulation of Ac-p65 and actived NF-κB in the spinal cord. Conclusion Intrathecal resveratrol can inhibit the development of neuropathic pain and suppress the activation of NF-κB signaling in SNL rats . The analgesic effect of resveratrol is implemented partly via increasing the level of SIRT1 and deacetylat-ing p65.

Objective To establish a arsenic cerium catalytic rate method for determination of urinary iodine,and increase the linear range of urinary iodine determination.Methods Standard series and urine samples after digestion treatment,were tested using dynamics function of spectrophotometer to record the curve of absorbance value (A) change with time (t) during arsenic cerium catalytic reaction for each measurement system,choice (A1,t1) and (A 2,t2) on this curve and calculating the reaction rate (v),v =(lgA1-lgA2)/(t2-t1).Through the determination of the standard series it could calculate regression equation of iodine concentration (C) with X:C =a ± bX,X =1 000 (v-v0),and the v0 is the reaction rate of reagent blank.Results (① C and X were positively correlated.The standard series linear range was 0-1 200 pμg/L and correlation coefficient r was higher than 0.999 1.The minimum detection limit was 3.9 μg/L (0.25 ml urine).②)Precision:5 urine samples (A,B,C,D,E) were selected within the range of 0-1 200 μg/L and the measured value were (72.3 ± 2.7),(148.2 ± 5.2),(210.5 ± 4.4),(562.7 ± 6.8),and (899.3 ± 8.0) μg/L.The relative standard deviation (RSD) was between 0.9％-3.8％.(③)Accuracy:4 samples (A,B,C,D) were measured for standard addition recovery test,recovery was between 94.2％-107.2％;urinary iodine standard material [the given values were (67.9 ± 9.0),(142.0 ± 10.0),(195.0 ± 10.0),(558.0 ± 17.0),(885.0 ± 28.0) μg/L] were determined and the results were in the range of uncertainty of the standard material.④Method contrast:with the national health standard method (method for determination of iodine in urine by arsenic cerium catalytic spectrophotometry) to determinate 120 urine samples,the results showed that there were 60 urine samples within 0-300 μg/L,60 urine samples were more than 300 μg/L.Then rate method was used to test the 120 urine samples.For the 60 samples within the scope of 0-300 μg/L,the determination results of the two methods were positively correlated (r =0.994,P ＜ 0.01);the results of the rate method were lower than those of the standard method and the difference was statistically significant (t =2.047,P ＜ 0.05).But the average deviation was only 2.1 μg/L,for the determination of urine iodine there was no practical significance;for the 60 samples higher than 300 μg/L,the determination results of the two methods were positively correlated (r =0.993,P ＜ 0.01) and the difference was not statistically significant (t =-1.092,P ＞ 0.05).Conclusions Arsenic cerium catalytic rate method has increased the linear range of urinary iodine determination.Using this method,the vast majority samples can be tested directly without dilution,thereby reducing the workload for determination of urine iodine.