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.

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.

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 .

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 measure serum surfactant protein (SP) A and D levels in patients with interstitial lung disease associated with rheumatoid arthritis (RA).Methods Serum SP-A and SP-D levels of RA,RA-ILD patients and healthy controls were assessed using a sensitive enzyme-linked immunosorbent assay (ELISA).The relationship between SP-A and SP-D and RA-ILD was analyzed.The serum SP-A and SP-Dpositive rate was calculated for the three groups.The correlation between SP-A and SP-D with RF,anti-CCP,antinuclear antibody,antikeratin antibody,anti-perinuclear factor,C-reactive protein,erythrocyte sedimentation rate,were analyzed.Mean value of groups were compared with variance analysis,Spearmam rank correlation test was used for correlation analysis.Results The levels of serum SP-A in RA-ILD patients and RA patients as well as in healthy controls were [ (51.2±9.2),(25.9±2.6),( 15A±0.3 ) μg/L] respectively.The level of serum SP-D of the three groups was [ ( 42.5 ±8.1 ),(20.8 ± 1.5 ),( 16.6±0.8 ) μg/L ] respectively.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were higher than those simple RA patients and healthy controls (P＜0.05).The levels of serum SP-A and SP-D in patients with RA were not significantly higher than those in healthy controls (P＞0.05).The positive rate of serum SP-A and SP-D in RA-ILD patients were significantly higher than those in simple RA patients and healthy controls.The positive rate of serum SP-D of RA-ILD patients was higher than that of SP-A.The levels of serum SP-A and SP-D in patients with RA complicated with ILD were correlated positively with age,C-reactive protein.The level of serum SP-D was correlated positively with RF,anti-CCP,antikeratin antibody.There was no correlation between the level of serum SP-A and SP-D with RA-ILD and antinuclear antibody,antiperinuclear factor,erythrocyte sedimentation rate.Conclusion The levels of serum SP-A and SP-D are correlated with RA-ILD and may be useful markers for ILD in patients with RA.These two paramenters may be helpful to early diagnosis of RA-ILD.The Serum SP-D levels are more sensitive in predicting the development of RA-ILD than other parameters and can help in assessing the severity of lung damage.

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.

Aim To explore the effect of knockdown spinal cord LCN2 by RNAi on the development of mor-phine tolerance in normal rats.Methods After suc-cessful intrathecal implantation, fourty-eight male Sprague-Dawley rats weighing 1 80 -220 grams were randomly divided into 4 groups (n =1 2):group I:control group,group II:morphine tolerance group, group Ⅲ:mismatch siRNA group,group IV:LCN2 siRNA group.The sixth day after intrathecal implanta-tion,rats were tested to ensure the position of cathe-ters,and it was recorded as d 0.On d 2 -8,rats were subcutaneously (s.c)injected of normal saline (NS) (group I)or morphine (group Ⅱ,Ⅲ,Ⅳ)1 0 μg· g -1 twice a day at 8:00 and 1 6:00.Before everyday s. c injection,rats were intrathecally injected of 1 0 μL DEPC solution (group Ⅰ,Ⅱ),1 0 μL DEPC solution containing 4 μg mismatch siRNA (group III)and 4 μg LCN2 siRNA solution (group IV).Paw withdrawal la-tencies to thermal stimuli (PWTL)were tested before morphine injection and 45 minutes after morphine in-jection on d 1 and d 9.The percentage of maximal pos-sible effect (% MPE)was calculated later.Animals were sacrificed on d 9 after the behavioral test and the lumbar enlargement segments of the spinal cord were removed for detecting the expression of phosphorylated-p38 mitogen-activated protein kinase (p-p38 MAPK) and LCN2 by Western blot and microglia marker Iba1 by immunofluorecence.Results On d 1 ,there was no significant difference in %MPE among four groups. On d 9,compared to group Ⅰ,%MPE was signifi-cantly reduced (P <0.05)while p-p38MAPK,LCN2 and Iba1 were markedly up-regulated in group Ⅱ andⅢ (P <0.05 ).On d 9,compared to group Ⅱ,%MPE was significantly increased while p-p38MAPK, LCN2 and Iba1 were markedly reduced in group IV (P<0.05).Conclusion Using LCN2 siRNA to knock-down spinal LCN2 relieves the development of mor-phine tolerance in normal rats possibly through inhibi-ting the activation of microglia and p38 MAPK in the spinal cord.