ObjectiveTo investigate the effects of Akt3 gene knockout on neuropathic pain behaviors induced by chronic constriction injury of sciatic nerve (CCI).MethodsExperiment was divided into two groups:Akt3 knockout group (Akt3-/-,n =12),wild type group (WT,n =12 ).Randomly numbered,the right sciatic nerve of mice were received the operation of chronic constriction injury.Paw withdrawal mechanical threshold (PWMT)and paw withdrawal thermal latency (PWTL) were tested on day 1 before operation and day 1,3,5,7,10,14,17,21 afterCCI.ResultsThe basic values of PWMT(right:(1.09±0.20)g,(1.17±0.22)g;left:(1.17±0.15)g,(1.22±0.23)g,P＞0.05) andPWTL(right:(6.18±1.11)s,(6.20±1.25)s;left:(5.82±0.91)s,(5.92± 1.71 ) s,P ＞ 0.05 ) had no statistically significant differences between two groups.On day 1 after operation,compared with basic values,the PWMT and PWTL of the right paw in both Akt3-/- group and WT group decreased significantly (P ＜ 0.05 ),and at least lasted up to day 21.The PWMT( 3d:(0.42 ± 0.22 ) g,(0.72 ± 0.36) g ; 17d:(0.29 ±0.15)g,(0.49 ±0.19) g;21d:(0.27 ±0.18)g,(0.56 ±0.15)g,P＜0.05) and PWTL(5d:(2.43 ±0.68)s,(3.13±0.52)s;17d:(2.43±1.26)s,(3.84±1.29)s ;21d:(2.14±1.23)s,(4.07±1.26)s,P＜0.05 ) of the right paw in Akt3-/- group was significantly lower than those in WT group.The PWMT and PWTL of the left paw in Akt3-/- group and WT group had no obvious differences (P ＞ 0.05 ). However.compared to left paw,the PWMT and PWTL of the right paw of the two groups were obviously lower (P ＜ 0.05 ).ConclusionThe neuropathic pain induced by CCI increased in Akt3 gene knockout mice.

Objective To explore the effect of pre-treatment of subcutaneous injection of ketamine on remifentanil induced hyperalgesia and K+/Cl-cotransporter 2,KCC2) expression on spinal cord of rats.Methods60 male adult SD rats were randomly divided into five groups(n=12 in each group):control group (group C),the incision group(group I),the incision plus remifentanil group(group I+R),the incision plus ketamine group(group I+K) and the incision plus remifentanil and ketamine group(group I+R+K).Mechanical withdrawal threshold (MWT) was evaluated at 24 hours before incision(T0),2 hours,6 hours,24 hours and 48 hours after incision(T1~T4).The lumbar spinal cords of rats were taken out at T4 time point and the KCC2 detected was detected by immunofluorescence analysis and western blot analysis.ResultsCompared with group C(T1(14.5±1.7)g,T2(14.2±1.1)g,T3(13.9±1.8)g,T4(14.2±1.1)g),MWT of other groups at T1 (I(5.6±0.8)g,I+R(3.2±1.0)g,I+K(6.8±1.7)g,I+R+K(5.1±1.6)g),T2 (I(6.9±1.0)g,I+R(4.3±1.2)g,I+K(8.0±1.4)g,I+R+K(6.2±1.5)g),T3 (I(7.6±0.9)g,I+R(5.4±1.1)g,I+K(10.3±1.2)g,I+R+K(7.1±1.1)g),T4 (I(8.9±1.1)g,I+R(7.5±1.4)g,I+K(11.3±1.2)g,I+R+K(8.3±1.2)g)and the expression of KCC2 at T4 decreased (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+R (T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g) decreased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 decreased significantly (P<0.05).Compared with group I(T1(5.6±0.8)g,T2(6.9±1.0)g,T3(7.6±0.9)g,T4(8.9±1.1)g),MWT of group I+K (T1(6.8±1.7)g,T2(8.0±1.4)g,T3(10.3±1.2)g,T4(11.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).Compared with group I+R(T1(3.2±1.0)g,T2(4.3±1.2)g,T3(5.4±1.1)g,T4(7.5±1.4)g),MWT of group I+R+K (T1(5.1±1.6)g,T2(6.2±1.5)g,T3(7.1±1.1)g,T4(8.3±1.2)g) increased at all time points after incision (T1~T4)(P<0.05) and the expression of KCC2 at T4 increased (P<0.05).ConclusionPre-treatment of subcutaneous injection of ketamine can reduce the hyperalgesia of rats induced by remifentanil and reduce the inhibition of KCC2 expression on dorsal horn of spinal cord.

Objective To evaluate long-term efficacy of microwave hyperthermia combined with chemoradiotherapy for nasopharyngeal carcinoma (NPC) with cervical lymph node metastasis. Methods A total of 154 patients with stage N2 or N3 NPC ('92 staging system) were randomized into two groups:microwave hyperthermia combined with chemoradiotherapy (Group A, 76 cases) and chemoradiotherapy alone (Group B, 78 cases). Both of the two groups received 1 -2 cycles of chemotherapy of cisplatin and 5-fluorouracil, followed by conventional radiotherapy of 70 - 78 Gy in 35 - 39 fraction to the nasopharynx and 68 -72 Gy in 34 - 36 fractions to the neck. Group A received microwave hyperthermia to the metastatic cervical nodes at the beginning of radiotherapy. The hyperthermia was given as 45 min every time, twice a week for 8 - 14 times totally. Results The 5-year complete response rates of cervical lymph nodes in group A and B were 97% and 77% (x2 = 14. 24,P<0. 01). The distant metastasis rates in the two groups were 37% and 44% (x2 = 0. 73, P > 0. 05). The disease-free survival rates were 51% and 21% (x2 = 15.91, P <0. 01). The 5-year overall survival rates were 59% and 41%, respectively (x2 = 5.09, P < 0. 05).Conclusions For patients with stage N2 or N3 NPC, microwave hyperthermia combined with chemoradiotherapy can improve the complete response, disease-free survival and overall survival.

Objective To investigate the effects of Aktl gene knockout on pain behavior induced by chronic constriction injury model of sciatic nerve (CCI).Methods C57BL/6 male mice were randomly divided into Akt1 knockout group (KO group,n=12),wild type group(WT group,n=12).All mice were made model of CCI in the right sciatic nerve.Each mouse received tests of the paw withdrawal mechanical threshold (PWMT) and the paw withdrawal thermal latency(PWTL) at the times of 1d before and 1 d,3 d,5 d,7 d,10 d,14 d,17 d,21 d after surgery.Results For both KO group and WT group,the basic values of PMWT(right(0.89±0.15)g,(0.87±0.15)g; left(0.97±0.19) g,(1.05±0.14) g,P＞0.05) and PWTL(right (7.64±0.71) s,(7.56±0.68) s ;left: (7.67±0.6) s,(7.64±0.64) s,P＞0.05) showed no significantly statistical difference.Compared with WT group and the basic value,PWMT and PWTL were significantly decreased after surgery in KO group (P＜0.05).The PWMT and P WTL of the left paw in KO group and WT group had no obvious statistical difference (P＞0.05).However,the PWMT and PWTL of the right paw significantly decreased in the two groups compared with left paw.Conclusion h aggravates the neuropathic pain induced by CCI in mice when the Akt1 gene was knocked out.

ObjectiveTo prospectively investigate the impact of short-time response on survival of concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC). Methods From Jan.2003 to Oct.2010,201 patients with pathologically or cytologically proven stage Ⅳ NSCLC were included.All patients received platinum-based chemotherapy.Of the 167 patients eligible for analysis,the median number of chemotherapy were 4 cycles.The median dose for planning target volume (PTV) of thoracic primary tumor was 63 Gy.Response was scored according to WHO criteria. Survival was calculated by Kaplan-Meier method and compared using the Logrank. Cox regression model were used to examine the effect of response on overall survival.ResultsThe follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years' follow-up.For the 167 patients eligible for analysis,the CR,PR,NC and PD rate of primary tumor was 5.4％,65.9％,21.0％ and 7.7％,respectively.The effective group ( CR + PR) and ineffective group ( NC + PD) was 71.3％ and 28.7％,respectively.The median survival time (MST) for patients with CR,PR,NC and PD was 22.6,13.4,8.8 and 4.8 months,respectively ( χ2 =44.79,P =0.000).The MST for effective and ineffective group was 13.9 and 7.6 months,respectively in the whole group ( χ2 =8.3 0,P =0.004 ),12.1months and 7.3 months in those treated with 2 - 3 cycles chemotherapy ( χ2 =7.71,P =0.007 ),and 13.9months and 7.9 months in those treated with 2 -5 cycles chemotherapy and radiation dose to PTV ≥36 Gy ( χ2 =4.00,P =0.045 ).No significant MST difference was detected between patients of effective group and ineffective group treated with 4 -5 cycles chemotherapy ( χ2 =0.67,P =0.413),or those treated with 4 -5 cycles of chemotherapy and radiation dose to primary lesion ≥36 Gy (χ2 =0.00,P =0.956).Multivariate analysis showed that 4-5 cycles of chemotherapy and CR and PR achieved in primary tumor (β =0.182,P=0.041 ) were independent favorable factors for survival. Conclusion CCTTRT can improve local control,and prolong the survival time for Stage Ⅳ NSCLC.

Objective To evaluate the preoperative liver function and prognosis of laparoscopic cholecystectomy (LC) in patients with cirrhosis,using the Child-Turcotte-Pugh classification and the model for end-stage liver disease(MELD) score.Methods From January 2009 to June 2013,973 patients who were admitted to the Department of General Surgery of our hospital and the HuiZhou Municipal Central Hosptial were studied.Of the 373 patients with cirrhosis,38 patients were excluded because of Child C,MELD ＞ 30,or laparotomy.The remaining 335 patients who received laparoscopic cholecystectomy were randomly divided into two groups The Child grade and MELD score were retrospectively analyzed.Results There was no significant difference in intraoperative hemorrhage between the Child A group [(106 ± 11) ml] and the Child B group [(109 ± 11) ml] (P ＞ 0.05).The R ＜ 14 scores in the MELD group [(58 ± 15) ml] was significantly lower than that in the R≥ 14 group [(120 ± 28) ml] (P ＜ 0.01).There was no significant difference in postoperative complications between the Child group A (10 cases,12％) and the Child group B (17 cases,21％) (P ＞0.05).There was a significantly lower incidence in the R ＜ 14 scores in the MELD group (10 cases,12％) than the R ≥ 14 group (27 cases,33％) (P ＜ 0.05).There was also no significant difference in the hospital stay between the Child A group (9 ± 1) and the Child B group (10 ± 2)(P ＞0.05) ; the R ＜ 14 score of the MELD group (7 ± 1) was significantly less than that of the R≥ 14 group (11 ±2) (P ＜0.01).There was no significant difference in the cost of hospitalization between the Child A group (1.337 ± 0.063) and the Child B group (1.359 ± 0.089) (P ＞ 0.05) ; the R ＜ 14 group (MELD score 1.108 ± 0.123) was significantly less than that of the R ≥ 14 group (1.568-± 0.117)(P ＜ 0.01).Conclusion Compared with the Child-Turcotte-Pugh classification,the MELD score was more scientific,objective and accurate in judging the preoperative liver function.It helped to predict the amount of intraoperative hemorrhage and postoperative morbidity,reduced hospital stay and hospitalization expenses.Therefore,the MELD scoring system more objectively guided the treatment of patients with cholecystitis with cirrhosis.

Objective To evaluate the importance of three-dimensional radiotherapy for elderly patients of stage Ⅳ non-small cell lung cancer (NSCLC).Methods Comparing with treatment outcome of ≥65 years 67 patients and ＜ 65 years 134 patients using concurrent chemotherapy and thoracic threedimensional radiotherapy during 2003 to 2010 years.Survival analysis was taken by Kaplan-Meier method.The multivariate prognosis was analyzed by Cox model.Results The follow-up was 97.8％.The percentage of ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy were 30％ and 55％,and with 42％ and 49％ patients with radiotherapy ≥63 Gy.The median survival time (MST) were 17 months and 14 months (x2 =0.76,P =0.384) for ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy concurrent ≥63 Gy radiotherapy respectively.The MST and 1-,2-,3year overall survival rate were 17 months and 8 months,65％ and 23％,30％ and 13％,24％ and 9％(x2 =7.90,P =0.005) for whole groups patients treated with chemotherapy concurrent ≥63 Gy and ＜ 63 Gy radiotherapy.And the MST of patients ≥ 63 Gy was significantly longer than those with ＜ 63 Gy either concurrent chemotherapy any cycles (x2 =9.54,P =0.023).The MST were 14 months and 8 months (x2 =1.82,P=0.178),17 months and 17 months (x2 =0.47,P=0.492) for ≥ 65 years and ≥ 63 Gy radiotherapy patients accepted with concurrent 4-5 cycles and 2-3 cycles chemotherapy concurrent respectively.Multivariate analysis showed local response (β =0.600,P =0.003) and numbers of tumor metastasis (β =0.670,P =0.040) were independent factors for survival.Conclusions For a part of elderly patients of stage Ⅳ NSCLC,concurrent chemotherapy and thoracic three-dimensional radiotherapy can prolong survival time with acceptable toxicity.Perhaps radiotherapy is more important.

Objective To prospectively evaluate the survival of different metastasis organs with concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC).Methods Two hundred and one patients of stage Ⅳ NSCLC were enrolled from January,2003 to July,2010.Of the 182 patients eligible for analysis,The number of patients with single-organ metastasis or multiple-organ metastasis was 107 and 75,respectively.Patients were treated by platinum-based chemotherapy,the median number of cycle was 4.The median dose to planning target volume of primary tumor (DTPTv) was 63 Gy.Survival was calculated by Kaplan-Meier method and compared using the Logrank.Results The follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years'follow up.Of 182 patients,the 1-,2-,and 3-year overall survival (OS) rate and median survival time (MST) was 41.0％,17.0％,10.0％ and 10.5 months,respectively ;with single-organ metastasis and multi-organ metastasis were 50％,20％,14％ and 13 months and 29％,12％,0％ and 8.5 months ( x2 =10.10,P =0.001 ),respectively; compared with multi-organ metastasis,the 1-,2-,and 3-year OS arte and MST of patients with bone,lung metastasis only was 58％,25％,16％ and 14 months (x2 =10.42,P=0.001 ) and 49％,21％,21％ and 11 months (x2 =6.39,P=0.011 ) respectively;patients with brain metastasis only did not show advantage of survival comparing with patients with multi-organ metastasis (49％,8％,0％ and 12 months and 29％,12％,0％ and 8 months,respectively;x2 =0.71,P =0.401 ) ;the 1-,2-,and 3-year OS rate and MST was 63％,23％,19％ and 15 months and 42％,15％,0％ and 10 months,respectively for patients with single-organ metastasis and multi-organ metastasis patients who accepted 4 - 5 cycles of chemotherapy ( x2 =6.47,P =0.011 ) ; for patients under the same metastasis and 4 - 5 cycles of chemotherapy,no matter whether single-organ or multiple-organ metastases,the 1 -,2-,3-year OS rate and MST of patients with enough radiotherapy on DTPTV ≥63 Gy were better than patients without enough radiotherapy ( DTPTV ＜ 63 Gy ) ( 71％,25 ％,25％ and 16.8 months and 33％,17％,0％ and 10.5 months,respectively;x2 ＝4.73,P =0.030 ;54％,21％,0％ and 14.3 months and 29％,10％,0％ and 7.6 months,respectively,x2 =8.16,P =0.004).The MST of liver metastases was 6 months,there was significantly difference when comparing with non liver matastasis ( x2 =17.21,P =0.000).Conclusions It is very important to treat stage Ⅳ NSCLC with CCTTRT,especially patients with single-organ metastasis.Liver metastases is a unfavorable prognostic factor.

Objective To investigate the effects of intrathecally cyclic AMP response element-binding protein(CREB) antisense oligodeoxynucleotide (ODN) on neuropathic pain behaviors.Methods Using mouse model of neuropathic pain induced by chronic constriction injury of sciatic nerve (CCI),24 male C57BL/6 mice successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into 4groups(n=6):Saline group(NS),CREB sense ODN group(S),CREB missense ODN group(M),CREB antisense ODN group(A).Mice in NS,S,M and A were intrathecally treated with Saline 5μ l,Sense ODN 5μg/5μl,Missense ODN 5μg/5μl and Antisense ODN 5μg/Sμl once daily on day 1 ～6 after CCI respectively.Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency(PWTL) were tested on day 1 before CCI and day 1,3,5,7,10,14,17,21 after CC(I).Results Mice in A group maintained the pain thresholds in the baseline and lasted at least 7 days after CCI ( 7 d,PWMT:( 0.81 ± 0.20 ) g vs ( 1.00 ± 0.19 ) g,P ＞ 0.05 ;PWTL:(5.96 ± 0.69) s vs (6.93 ± 1.08 ) s,P ＞ 0.05 ).The withdrawal thresholds in the ipsilateral hind paws of the mouse were significantly lower than baseline in A group on day 10 after CCI( 10 d,PWMT:(0.56 ±0.19)g vs (1.00±0.19)g,P＜0.05; PWTL:(3.93 ±0.28)s vs (6.93 ± 1.08)s,P＜0.05).Compared with NS group ( 10 d,PWMT:(0.56 ±0.19)g vs (0.37 ±0.08)g,P＜0.05; PWTL:(3.93 ±0.28)s vs (3.14 ±0.45)s,P＜0.05),S group,M group,the withdrawal thresholds of A group was significantly elevated on day 10 after CCI.These effects lasted up to at least day 21 after CCI.Conclusion Intrathecally treated with CREB antisense ODN in the development of neuropathic pain induced by CCI completely improved pain behaviors during the course of injection,and the effects of relief pain lasted at least 15d after no injection.