Toll-like receptor 4uclear factor-kappa B pathway is involved in radicular pain by encouraging spinal microglia activation and inflammatory response in a rat model of lumbar disc herniation

Background@#Lumbar disc herniation (LDH) is a common cause of radicular pain, but the mechanism is not clear. In this study, we investigated the engagement of toll-like receptor 4 (TLR4) and the nuclear factor-kappa B (NF-κB) in radicular pain and its possible mechanisms. @*Methods@#An LDH model was induced by autologous nucleus pulposus (NP) implantation, which was obtained from coccygeal vertebra, then relocated in the lumbar 4/5 spinal nerve roots of rats. Mechanical and thermal pain behaviors were assessed by using von Frey filaments and hotplate test respectively. The protein level of TLR4 and phosphorylated-p65 (p-p65) was evaluated by western blotting analysis and immunofluorescence staining. Spinal microglia activation was evaluated by immunofluorescence staining of specific relevant markers. The expression of proand anti-inflammatory cytokines in the spinal dorsal horn was measured by enzyme linked immunosorbent assay. @*Results@#Spinal expression of TLR4 and p-NF-κB (p-p65) was significantly increased after NP implantation, lasting up to 14 days. TLR4 was mainly expressed in spinal microglia, but not astrocytes or neurons. TLR4 antagonist TAK242 decreased spinal expression of p-p65. TAK242 or NF-κB inhibitor pyrrolidinedithiocarbamic acid alleviated mechanical and thermal pain behaviors, inhibited spinal microglia activation, moderated spinal inflammatory response manifested by decreasing interleukin (IL)-1β, IL-6, tumor necrosis factor-α expression and increasing IL-10 expression in the spinal dorsal horn. @*Conclusions@#The study revealed that TLR4/NF-κB pathway participated in radicular pain by encouraging spinal microglia activation and inflammatory response.

Toll-like receptor 4uclear factor-kappa B pathway is involved in radicular pain by encouraging spinal microglia activation and inflammatory response in a rat model of lumbar disc herniation

Background@#Lumbar disc herniation (LDH) is a common cause of radicular pain, but the mechanism is not clear. In this study, we investigated the engagement of toll-like receptor 4 (TLR4) and the nuclear factor-kappa B (NF-κB) in radicular pain and its possible mechanisms. @*Methods@#An LDH model was induced by autologous nucleus pulposus (NP) implantation, which was obtained from coccygeal vertebra, then relocated in the lumbar 4/5 spinal nerve roots of rats. Mechanical and thermal pain behaviors were assessed by using von Frey filaments and hotplate test respectively. The protein level of TLR4 and phosphorylated-p65 (p-p65) was evaluated by western blotting analysis and immunofluorescence staining. Spinal microglia activation was evaluated by immunofluorescence staining of specific relevant markers. The expression of proand anti-inflammatory cytokines in the spinal dorsal horn was measured by enzyme linked immunosorbent assay. @*Results@#Spinal expression of TLR4 and p-NF-κB (p-p65) was significantly increased after NP implantation, lasting up to 14 days. TLR4 was mainly expressed in spinal microglia, but not astrocytes or neurons. TLR4 antagonist TAK242 decreased spinal expression of p-p65. TAK242 or NF-κB inhibitor pyrrolidinedithiocarbamic acid alleviated mechanical and thermal pain behaviors, inhibited spinal microglia activation, moderated spinal inflammatory response manifested by decreasing interleukin (IL)-1β, IL-6, tumor necrosis factor-α expression and increasing IL-10 expression in the spinal dorsal horn. @*Conclusions@#The study revealed that TLR4/NF-κB pathway participated in radicular pain by encouraging spinal microglia activation and inflammatory response.

Ulinastatin ameliorates liver dysfunction during perioperative period of hepatectomy in liver cancer patients / 中华普通外科杂志

Objective To investigate the protective effect of Ulinastatin on liver function after precise hepatectomy.Methods Fifty-six patients of hepatocellular carcinoma undergoing hepatectomy were divided into Ulinastatin group (U group,28 cases) and control group (group C,28 cases).Results There were no significant differences in liver function and inflammatory factors between the two groups on the 1 st day before the surgry (P＞ 0.05).Serum ALT levels of (231 ±29;140 ±21;56 ± 13;42 ±6) U/L,were lower than group C (267 ± 29;158 ± 23;69 ± 18;53 ± 8) U/L,all P ＜ 0.05;serum AST levels were (175 ±23;94 ±25;47 ± 16;36 ± 8) U/L,lower than group C (191 ± 17;139 ± 16;64 ± 15;46 ±11) U/L,all P＜0.05;Serum TBIL levels were (30 ±6;39 ±9;31 ±9;21 ±6) μmol/L,lower than group C (34 ± 6;46 ± 7;35 ± 8;26 ± 7) μmol/L,all P ＜ 0.05;Serum hs-CRP levels were (52 ± 22;112 ±23;71 ± 16;42 ± 13) rg/L,lower than group C (69 ±23;129 ±25;72 ± 15;49 ± 15) mg/L,all P ＜ 0.05;serum IL-1 β levels were (7.8 ± 0.9;11.1 ± 1.5;8.9 ± 1.6;5.6 ± 0.9) pg/ml,lower than group C (9.2±1.3;13.0 ±2.1;11.1 ±1.4;7.3 ±0.8) pg/rml,all P ＜0.05;Serum IL-6 levels were (187 ±30;76 ±25;46 ± 15;26 ±8) pg/ml;lower than group C(260 ±36;92 ± 16;53 ± 11;30 ±8) pg/ml,all P ＜ 0.05;Serum TNF-α levels were (17 ± 4;12.0 ± 2.4;8.6 ± 2.4;6.7 ± 2.0) pg/ml,lower than group C (25 ± 4;18.7 ± 3.3;15.0 ± 3.1;9.6 ± 1.7) pg/ml,all P ＜ 0.05.Conclusion Ulinastatin protects liver function in perioperative patients undergoing precise hepatectomy.

Significance of p16/Ki-67 double immunocytochemical staining in cervical cytology ASCUS, LSIL,and ASC-H / 中华妇产科杂志

Objective To investigate the application value of p16/cell proliferation associated nuclear antigen (Ki-67) double-staining and human papillomavirus mRNA in the cytological screening.Methods Two hundred and fifty-one cases who suffered from atypical squamous cell of undetermined significance (ASCUS),low-grade squamous intraepithelial lesion (LSIL),atypical squamous cell-cannot exclude high-grade squamous intraepithelial lesion (ASC-H) in ThinPrep cytologic test (TCT) were collected in Peking University First Hospital between October 2015 and March 2016.And p16/Ki-67 double-staining and hybrid capture Ⅱ (HC-Ⅱ) detection were performed on the cervical cells.The result was compared with the pathological result of colposcope guided biopsy.All statistical analysis was completed by Stata 12.0 statistical software analysis.The results of diagnostic tests were described by using the sensitivity,specificity,positive predictive value,negative predictive value,and the area under the receiver operating characteristic (ROC) curve.Results (1) One hundred and eight cases of liquid based cytology diagnosis of ASCUS patients,the positive rate of p16/Ki-67 was 13.9％ (15/108),102 cases of liquid based cytology diagnosis of LSIL patients,the positive rate of p16/Ki-67 was 21.6％ (22/102),41 cases of liquid based cytology diagnosis of ASC-H patients,the positive rate of p16/Ki-67 was 39.0％ (16/41),compared amongthree groups,the difference was statistically significant (x2=78.516,P＜0.05);cervical exfoliated cells p16/Ki-67 expression rate was 13.0％ (28/215) in cervical low-grade lesions [cervical intraepithelial neoplasia (CIN)Ⅰ],which was 69.4％ (25/36) in high level lesions (CIN Ⅱ-Ⅲ),the difference was statistically significant (x2=7.932,P＜0.05).(2) The specificity of p16/Ki-67 detection and diagnosis were higher than those of HC-Ⅱ in ASCUS,LSIL,and ASC-H (89.8％ vs 71.4％,83.3％ vs 15.6％,88.9％ vs 40.7％;all P＜0.05),meanwhile,the positive predictive value of p16/Ki-67 detection and diagnosis exceed those of HC-Ⅱ in ASCUS,LSIL,and ASC-H (33.3％ vs 26.3％,31.8％ vs 12.6％,81.3％ vs 38.5％;all P＜0.05).Moreover,the ROC curve of p16/Ki-67 were bigger than those of HC-Ⅱ in ASCUS,LSIL,and ASC-H (0.799 vs 0.696,0.708 vs 0.531,0.909 vs 0.561;all P＜0.05).Conclusion For patients with cytological diagnosis of ASCUS,LSIL,and ASC-H,p16/Ki-67 double staining method could be used as an effective method to assist in the diagnosis of high-grade cervical lesions,and the screening efficiency is superior to that of high-rist HPV.

Comparison of clinical pathological characteristics in ovarian preserving patients with stageⅠB1 cervical adenocarcinoma and squamous cell carcinoma / 北京大学学报(医学版)

Objective:To analyze the risk and prognostic of patients with stage ⅠB1 cervical adenocar-cinoma.Methods:The clinical data of 139 patients with stage ⅠB1 cervical adenocarcinoma treated at Department of Gynecology and Obstetrics in Peking University First Hospital from August 1994 to April 2015 were retrospectively reviewed,which included 38 cases of cervical adenocarcinoma and 101 cases of cervical squamous cell carcinoma.A comparison was made between ovarian preserving group and bilateral oophorectomy group,in order to justify the risk and prognosis of ovarian preserving patients.Results:The 5-year cumulative survival rate of stage ⅠB1 cervical adenocarcinoma and squamous cell carcinoma were 89.1% and 92.9% respectively with significant difference (P =0.034).One ovarian metastasis case was observed among the 32 cervical adenocarcinoma patients of bilateral oophorectomy,while another ovarian metastasis case was observed among 54 cervical squamous cell carcinoma patients of bila-teral oophorectomy.The ovarian metastasis rate was 3.1% (1 /32)and 1.8 % (1 /54)respectively with no statistical difference (P =0.574).The cumulative 5-year survival of 6 ovarian preserving patients with cervical adenocarcinoma was 80.1%,while that of 47 ovarian preserving patients with cervical squa-mous cell carcinoma was 94.6% (P =0.127).There was no statistical difference between the survival curve of the two groups.Conclusion:The prognosis of stageⅠB1 cervical adenocarcinomas was somewhat poorer than that of cervical squamous cell carcinoma.However it was still reasonable to perform ovarian preservation among young patients of stageⅠB1 cervical adenocarcinoma with no high risk factors.

Association of ApoB/ApoA-1 Ratio with Insulin Resistance in Patients with Non-alcoholic Fatty Liver Disease / 昆明医科大学学报

Objective This study aims to investigate the association of apoB/apoA-1 ratio with insulin resistance (IR) in patients with non-alcoholic fatty liver disease (NAFLD) . Methods A total of 224 patients with NAFLD and 166 healthy subjects were enrolled as NAFLD group and control group. Weight, height and blood pressure were recorded. Serum levels of fasting blood glucose (FPG), insulin (Fins), lipids, glycated hemoglobin (HbA1c) were measured. Homeostasis model assessment-insulin resistance (HOMA-IR) indices were calculated. Results Compared with control group, NAFLD group had higher apoB/apoA-1 ratio (0.76 ± 0.28 vs 0.61 ± 0.26) and HOMA-IR (2.43 ± 1.68 vs 1.86 ± 1.61) . Spearman correlation analysis showed that in NAFLD group, HOMA-IR positively correlated with age, body mass index (BMI), triglycerides (TG), low-density lipoprotein cholesterol (LDL-c), apoB/apoA-1 ratio (r =0.34, P < 0. 05) and HbA1c, and negatively correlated with high-density lipoprotein cholesterol (HDL-c) . Multiple linear regression analysis revealed that apoB/apoA-1 ratio was still associated with HOMA-IR in NAFLD group after adjustment for age and BMI. Conclusion The apoB/apoA-1 ratio is closely associated with IR in patients with NAFLD. ApoB/apoA-1 ratio may play a role in the development of IR in NAFLD.

Results of different postoperative adjuvant therapies for stage Ⅰ b-Ⅱ a cervical carcinoma with risk factors / 中华妇产科杂志

Objective To investigate the effects of postoperative adjuvant chemotherapy (CT) and chemoradiotherapy (CRT) or radiotherapy(RT) for Ⅰ b-Ⅱ a cervical cancer with risk factors.Methods From March 1995 to June 2010,there were 137 patients underwent radical hysterectomy and systematic pelvic lymphadenectomy for stage Ⅰ b-Ⅱ a cervical cancer admitted at Peking University First Hospital.These patients had risk factors,intermediate risk factors including bulky tumor (＞4 cm),lymph vascular space invasion,deep stromal invasion; high risk factors including positive surgical margin,parametrial invasion,lymph node involvement.Of the all patients,79 cases of them were treated with CT,58 of them were treated with RT or CRT.The 5-year survival and prognosis factors were analyzed retrospectively,the prognosis was compared between two adjuvant therapy groups.Results The univariate analysis shown that types of pathology,different grade of risk factors,stroma invasion and lymph node involvement were prognostic factors of 5-year overall survival Patients with squamous cell carcinoma,intermediate risk factors,no parametrial invasion,and no lymph node involvement had better prognosis (P ＜ 0.05).Whether patients with high-risk factors or intermediate-risk factors,the 5-year overall survival and 3-year disease-free survival had no difference between CT and RCT or RT groups respectively.Cox regression multivariate analysis of survival indicated that clinical stages,types of histology,different grade of risk factors were independent prognostic indicator.Patients with early stage,squamous cell carcinoma,intermediate risk factors had better prognosis.Univariate and multivariate analysis indicated that different postoperative adjuvant therapies had no effects on the prognosis.The 5-year overall survival was 88.6％ in patients treated with CT,and 89.7％ in patients treated with RT or CRT (P =0.455).Conclusion There are equivalent therapeutic results between CT and RT or CRT for patients with risk factors after radical surgery,CT may be as one choice of postoperative adjuvant therapy for stage Ⅰ b-Ⅱ a cervical carcinoma with risk factors.

Construction and verification of the prognostic model of ovarian serous adenocarcinoma / 肿瘤

Objective:To analyze the prognosis-related factors of ovarian serous adenocarcinoma in order to set up a prognostic model of serous adenocarcinoma and verify the effectiveness of the model as prognostic clinical criteria.Methods:The clinical, patholo-gical and follow-up data from 181 training samples with ovarian serous adenocarcinoma in Peking University First Hospital during January 1995 to December 2003 and another 42 detection samples with ovarian serous adenocarcinoma in Beijing Cancer Hospital during January 1999 to December 2005 were analyzed retrospectively. Kaplan-Meier univariate analysis was used to screen out prognostic factors; COX univariate and multivariate analyses were used to determine the risk coefficient of each factor and different layers in each factor; Pearson rank correlation analysis was used to identify the correlation of each factor. The prognostic model of ovarian serous adenocarcinoma was established on the conversion of risk coefficient to prognostic score and receiver operating characteristic (ROC) analysis was used to determine the cut-off value. The clinical data were collected to verify the sensitivity and specificity of the prognostic model based on the 3-year survival rate and Ki67 value.Results:The survival rate of patients with ovarian serous adenocarcinoma correlated with 6 factors including FIGO stage, histological grade, residual size, metastasis of lymph nodes, general condition after chemotherapy, and serum CA125 levels. The postoperative chemotherapy was an independent factor for prognosis. The prognostic model directly reflected the survival probability of classical COX risk ratio model, with higher score indicating lower survival probability and poorer prognosis. If combining prognostic score with Ki67 the sensitivity and specificity reached 64.7% and 96.0%, respectively.Conclusion:FIGO stage, histological grade, residual size, metastasis of lymph nodes, postoperative chemotherapy and serum CA125 were the prognostic factors of patients with ovarian serous adenocarcinoma. The prognostic model of ovarian serous adenocarcinoma can moderately reflect the actual survival status, and combination with Ki67 will increase the sensitivity and specificity.

The values of serunl human epididymis secretory protein 4 and CA125 assay in the diagnosis of ovarian malignancy / 中华妇产科杂志

Objective To evaluate the value of human epididymis secretory protein 4(HE4)and CAl25 in the diagnosis of ovariall malignancy.Methods HF4 and CA125 in the serum specimens of malignant ovarian tumor group(30 cases),benign ovarian diseases(110 cases;45 benign ovarian tumor,57endometriotic diseases and 8 pelvic inflammation were included) and healthy women group( 137 cases)were assayed double blindly . The levels and the diagnosis efficiency of the HE4 and CA125 were analyzed.Results (1) The median levels of HE4 and CA125 were significantly higher in malignant ovarian tumor group (244 pmoi/L and 601 kU/L respectively) than those of the benign ovarian diseases group( 32 pmol/L and 22 kU/L respectively)and healthy women group (32 pmoi/L and 11 kU/L respectively) (P =0. 000-0. 029). The median levels of CA125 were also higher in endometriotic diseases and pelvic inflammation groups(53 and 41 kU/L respectively) than those of benign ovarian tumor group and healthy women group (12 and 11 kU/L respectively;P = 0. 000-0. 031 ). (2) The positive rate of HE4 was lower than that of CA12s in malignant ovarian tumor group ( P = 0. 036 ). HE4 was negative in benign diseases and healthy women groups. But the positive rates of CA125 were 56. 1% and 5/8 respectively in endometriotic diseases and pelvic inflammation groups and there were significant differences compared with HE4( P =0. 000). (3)The HE4 assay had advantage over the CA125 assay in receiver operating characteristic-area under the curve (ROC-AUC) and sensitivity with a specificity of 100% when ovarian malignancy was compared with controls having benign diseases and healthy women, benign tumor or benign diseases groups respectively. The CA125 assay had advantage over the HE4 assay in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having healthy women group. (4) Combined assay of HE4 and CA125was better than CA125 alone when ovarian malignancy was compared with controls having any group. (5)Combined assay was better than HE4 alone in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having benign diseases and healthy women or healthy women groups. And combined assay was lower in the ROC-AUC and the sensitivity with specificity of 100% than HE4 when ovarian cancers were compared with controls having benign tumors or benign diseases groups respectively. (6) The diagnosis efficiency of the HE4 assay at the level 86 pmol/L determined in ROC curve with controls having benign diseases and healthy women group and at the 95% reference level 50 pmol/L of healthy women or 150 pmol/L recommended by the kit respectively was compared. The sensitivity of 50 pmol/L was 73% higher than 150 pmol/L and 86 pmoi/L, while the specificity and positive predictive value were lower ( P = 0. 002, P = 0. 000 ). The specificity, accuracy and positive predictive value of HE4 assay at the set point of 150 pmol/L and 86 pmol/L were 100%, 96% and 96%. The set point of 86 pmol/L had advantage over 150 pmol/L at the sensitivity of diagnosis, 70% and 63% respectively. But the positive predictive value was 95% lower than 150 pmol/L, being 100%. There was no significant difference( P =0. 883, P = 0. 883 ). Conclusions The specificity of HF4 assay is higher than CA125 assay in the diagnosis of ovarian cancer and HE4 combined with CA125 assay can improve the diagnoses. The set point of 150 pmol/L is advantageous for the accurate diagnosis, while the set point of 86 pmol/L is advantageous for the screening of malignant ovarian cancer.

Assessment of magnetic resonance imaging in preoperative staging of endometrial carcinoma / 北京大学学报(医学版)

Objective:To evaluate the usefulness of T2-weighed magnetic resonance images (MRI) in preoperative assessment of the depth of myometrial invasion and the staging of patients with endometrial carcinoma. Methods: 33 cases of endometrial carcinoma were examined by MRI before operation, and the pathological findings were compared with MRI findings.Results:The overall accuracy of T2-weighed magnetic resonance images in staging endometrial carcinoma was 84.85% (28/33). The accuracy for distinguishing stages Ⅰa,Ⅰb andⅠc was 84.62%(22/26). And the accuracy for localization of myometrial invasion (stageⅠc between stages Ⅰa+Ⅰb) was 96.15%(25/26). All the 3 cases with cervical invasions confirmed by pathological findings were accurately assessed by MRI. Conclusion:MRI is useful in assessing the depth of myometrial invasion and accurate staging in patients with endometrial carcinoma.

Establishment of a novel canine model of severe acute cholangitis / 中国普通外科杂志

Objective To establish a novel canine model of severe acute cholangitis(ACST).Methods Ten dogs were randomly divided into two groups:model group and control group.ACST models were established by placing T-tube in common bile duct and ligating the distal bile duct,then injecting E.coli trough T-tube and closing the T-tube.The changes of peripheral WBC,total bilirubin,ALT,AKP,IL-1?,TNF-? and endotoxin were observed pre-operatively and at 24 h,48 h,72 h,and 7 d post-operatively.The pathological changes of bile duct were observed when the dogs were sacrificed.Results There were no significant differences in all indices between two groups pre-operatively.Serum IL-1?,TNF-?,endotoxin and total bilirubin,ALT,AKP level in model group increased gradually and were higher than control group(P

A modified liver cirrhosis portal hypertension model in canines induced by subcutaneous injection of carbon tetrachloride / 中国普通外科杂志

Objective To find an ideal liver cirrhosis portal hypertension model in big animals suitable for surgical study.Methods Twenty canines were treated by subcutaneous injection of 60% CCl4 colza oil emulsion into the back,at a dose of 1.0-1.3 mL/kg every 10 days for a total of 6-8 times.At the same time,it was combined with the control of food intake.All canines were fed with rice mixed with 10% hog fat.The amount of rice was 15 g/kg per day from the first day to the forth day,but the amount of food was not controlled from the fifth day to the tenth day.During the process,the canines′ general condition was observed and the changes of hepatic functions such as ALT,TP,ALB,G and A/G were measured.The changes of liver morphology,the diameter and blood flow of portal vein were monitored with color Doppler.Every two weeks,a piece of hepatic tissue to observe the pathological change of liver was excised operatively.Results During the process,the body weight of the canines decreased continually;ALT increased during early and middle stages and decreased during advanced stage;TP and ALB always decreased,and A/G continuously decreased,but G did not change significamtly.Eight weeks after injection,liver became progressively smaller.its density was not well-distributed,and liver particles gradually became coarse,and liver capsule became rough.The diameter of portal veins became larger and the velocity of portal vein became slower.Liver cirrhosis with psudolobules was found on light microscopy from the tenth week to the twelfth week.Three canines died,with mortality of 15.0%.Conclusions It took 10-12 weeks to establish this liver cirrhosis portal hypertension models in canines by subcutaneous injection of CCl4.This method is convenient,has high success rate,and is suitable for use in surgieal research.