To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Decidua/metabolism ; Embryo Implantation/*physiology ; Endometrium/metabolism ; I-kappa B Proteins/*biosynthesis ; NF-kappa B/*biosynthesis ; Time Factors ; Uterus/*metabolism ; Uterus/physiology

Objective To establish a nude mice model for NB4/ShHMGA2 xenograft and explore the effect of HMGA2 knockdown on hematological malignancies. Methods NB4/ShHMGA2 or NB4/ShControl cell lines were established by transfecting the recombinant Lentivirus-HMGA2shRNA and the vacant Lentivirus-NC-marked into NB4 cells. The knockdown of HMGA2 was identified by RT-PCR and Western blot. Ten male BALB/c nude mice aged 4 ~ 5 weeks were equally divided into two groups. The mice irradiated by 4 Gy 60 Co were subcutaneously injected with 8 × 106 NB4/ShHMGA2 or NB4/ShControl cells into one side of axilla. The volumes of xenograft tumor were evaluated using the equation volume (mm3) = (L × W2)/2. The xenograft tumor section was detected by IHC with Ki-67 antibody. Results NB4 cell xenograft tumors developed in all mice of both the two groups. The NB4/ShHMGA2 cells in the nude mice grew at a lower rate than those in the controls. There were statistically significant differences in the volume and weight of xenograft tumor between the two groups [(1 484.25 ± 156.342)mm3 vs (3 228.674 ± 285.64)mm3, P < 0.05] and [(2 135.33 ± 198.05) mg vs (650.46 ± 85.12)mg, P < 0.05]. The Ki-67 protein level in NB4/ShHMGA2 cells xenografts was lower than that in the controls. Conclusion The knockdown of HMGA2 could inhibit proliferation of NB4 cells in NB4 cells xenograft tumor.

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.

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.

Aim Toinvestigatetheantagonisticeffect of intrathecal injection of carbenoxolone (CBX ) on neuropathic pain and its underlying mechanism.Meth-ods SixtymaleSprague-Dawleyratswererandomly divided into five groups (n =12 ):group I received sham surgery then treated with saline;group Ⅱ re-ceived SNT then treated with saline;groupⅢreceived SNT then treated with 0. 05 μg CBX;group Ⅳ re-ceived SNT then treated with 0. 5 μg CBX;group Ⅴreceived SNT then treated with 5 μg CBX.Treatment was undertaken with 10 μl volume as a single intrathe-cal injection on postoperative day 10.Mechanical with-drawl thresholds were measured 1 d before operation, 1,3,5,7 and 10 d after surgery,1 h before intrathe-cal administration,and 1 ,2,4,6 h after intrathecal administration.Lumbar spinal cord was obtained 2 h after intrathecal administration to determine the expres-sions of GFAP by immunohistology and TNF-α,IL-1βby ELISA in bilateral spinal dorsal horns.Results Comparedwiththeshamgroup,thebilateralMWTin group Ⅱ ~Ⅴ was significantly decreased.Compared with the MWT 1 h before intrathecal administration on day 10,the values at 1 ,2,4,6 h after administration of group Ⅱ and Ⅲ had no marked difference.The ip-silateral MWT in groupⅣhad no significant difference at 1,2,4 h after administration,the contralateral MWT was significantly increased,whereas GFAP and TNF-α,IL-1βwas significantly decreased in the spinal cord .In group Ⅴthe bilateral MWT was significantly improved at 1 ,2,4 h after administration,whereas GFAP and TNF-α,IL-1βwere significantly decreased inthespinalcord.Conclusions IntrathecalCBXcan inhibit the development of bilateral MWT.The analge-sic effect of CBX is implemented partly via suppressing the actation of GFAP and the realease of TNF-α,IL-1βin the spinal doral horn.

Objective To evaluate the role of expression of phosphodiesterase 4B (PDE4B) in the spinal cord in inflammatory responses in a rat model of neuropathic pain and the relationship with extracellular signal-regulated kinase (ERK).Methods Sixty healthy male Sprague-Dawley rats,weighing 180-220 g,in which intrathecal catheters were implanted at L5,6 interspace,were used.The location of catheters was confirmed 6 days later.The rats were randomly divided into 5 groups (n =12 each):sham operation group (group Sham),normal saline (NS) group,vehicle group (group Ⅴ),mismatch siRNA group (group siR-M),and PDE4B-siRNA group (group siR-B).Neuropathic pain was induced by ligation of L5 spinal nerve (SNL).In Sham group,the L5 spinal nerve was only exposed,but not ligated.Immediately after ligation and on 1,3,5,and 7 days after ligation,10 μl NS,10 μl NS,LipofectaminTM RNAiMAX,PDE4B-siRNA (2 μg/10 μl) encapsulated in mismatch siRNA and PDE4B-siRNA (2 μg/10 μl) encapsulated in LipofectaminTM RNAiMAX were injected intrathecally in Sham,NS,V,siR-M,and siR-B groups,respectively.The mechanical pain threshold was measured at 1 day before and 2,4,6 and 8 days after operation.After behavioral testing on 8th day after operation,the rats were sacrificed and the lumbar segment of the spinal cord was removed for determination of PDE4B protein,ERK and phosphor-ERK (p-ERK)expression,and TNF-α,IL-1β and IL-6 levels.Results Compared with Sham group,the mechanical pain threshold was significantly decreased at 2,4,6 and 8 days after operation in NS,V,siR-M and siR-B groups (P ＜0.05),and no significant change was found in the mechanical pain threshold at 2,4,6 and 8 days after operation (P ＞ 0.05) and the expression of p-ERK and PDE4B protein,and levels of TNF-α,IL-1β and IL-6 were increased at 2,4,6 and 8 days after operation in V and siR-M groups (P ＜ 0.05).Compared with NS group,the mechanical pain threshold was significantly increased,and the expression of p-ERK and PDE4B protein and levels of TNF-α,IL-1β and IL-6 were decreased at 2,4,6 and 8 days after operation (P ＜ 0.05),and no significant change was found in the parameters mentioned above in V and siR-M groups (P ＞ 0.05).Conclusion Up-regulation of the expression of PDE4B protein in the spinal cord is involved in the development of neuropathic pain in rats,which may be related to promoted phosphorylation of ERK in the spinal cord and enhanced inflammatory responses.

Objective To investigate the effect of intrathecal (IT) transplantation of different quantities of neural stem cells (NSCs) on neuropathic pain (NP) in rats.Methods Eighty-four adult pathogen-free male SD rats weighing 150-180 g were randomly divided into 7 groups ( n = 12 each) : group sham operation (S group) , NP group, NP+ NSCs 103 , 104 , 105 , 106 , 107 groups (N1-5 groups) . NP was induced by partial transection of right sciatic nerve. NSCs were transplanted into subarachnoid space in N1-5 groups. Paw withdrawal threshold to mechanical stimulation (MWT) and paw withdrawal latency to nociceptive thermal stimuli (TWL) were measured at 1 day before (baseline) and 1, 3, 7, 14 and 21 days after operation. Brain derived neurotrophic factor ( BDNF) expression in spinal dorsal horn and dorsal root ganglia (DRG) was detected by immuno-histochemistry and RT-PCR on the 7th and 21st day after operation. Results Partial transection of the sciatic nerve gradually reduced MWT and TWL after operation starting from day 1 until day 7 and 14 as compared with the baseline in group NP. IT NSC transplantation significantly increased MWT and TWL and expression of BDNF in spinal dorsal horn and DRG in a dese-dependent manner at day 7 after operation in N1-5 groups as compared with group NP. There were no significant differences in MWT and TWL and BDNF expression among N3, N4 and N5 groups at day 21 after operation.Conclusion The proper quantity of transplanted NSCs which are able to ameliorate NP induced by partial transection of sciatic nerve in rats is 105 .

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 investigate the effects of silenced Racl on invasion and migration in LOVo cells.Methods The expression of Racl mRNA and protein in colorectal cancer cells(including LoVo SW480.SW620.SW1116,HT29)were detected by RT-PCR and Western blot,respectively.The changes of cytoskeleton were observed in LoVO cells after transfected with Racl-shRNA.then invasion and migration were recorded respectively in LoVo cells after transfected with Racl-N17 and Racl-L61.Results Racl mRNA and protein were overexpressed in all selected colorectal cancer cells.Deletion of Racl decreased the cross-linked actin network and pseudopodia,and inhibited the invasion and migration in LoVo cells.The migration experiment showed that the migrated cells were higher in Racl-shRNA[(75±5)cells].Racl-N17 [(93±5)cells]and Racl-L61[(267±7)cells]groups compared with control group[(214±8)cells,P＜0.01,＜O.01 and＜0.05,resprectively].The invasion experimental study revealed that the migrated cells were higher in Racl-shRNA[(35±5)cells],Racl-N17[(42±5)cells]and Racl-L61[(86±7)cells] groups compared with control group[(73±6)cells,P＜0.01,＜0.01 and＜0.05,resprectively].Condusion Deletoin of Racl can inhibit the invasion and migration in LDVo cells.

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.