1.Results of open surgery or endoscopic choledocholithotomy in the treatment of hepatolithiasis and risk factors
Chenglin ZHU ; Qiang HUANG ; Chenhai LIU ; Fang XIE ; Qian YAO
Chinese Journal of General Surgery 2012;27(10):781-785
Objective To investigate the short and long-term outcomes and risk factors after open surgery or and endoscopic hepatolithotomy for hepatolithiasis. Methods A retrospective study was made on cases of hepatolithiasis who underwent hepatolithotomy by open surgery or endoscopically from Jan 2001 to Dec 2008.Of 254 patients,189 were followed-up including 127 after open surgery and 62 endoscopically.The univariate and multivariate analyses were performed to determine the risk factors. Results Complete stone clearance was achieved in 85.0 % ( 108/127 ) of open surgery including hepatecomy,61.3 % ( 38/62 )of endoscopic treatment.After a median follow-up period of 6.0 years (2.5 to 10.5 years),stone recurred in 32.8% (62/189) of patients,biliary cirrhosis in 7.4% ( 14/189),cholangiocarcinoma in 7(3.7% ),all cancer cases were dead with a mortality rate of 7(3.7% ).Bile duct stricture (OR:7.522,95% CI:2.642 -21.415),stones in both lobes (OR:11.630,95% CI:3.989 -33.912),and endoscopic treatment ( OR:21.374,95% CI:6.713 - 68.056 ) were independent risk factors ( P < 0.05 ) for incomplete stone clearance by unconditional Logistic regression analysis.In addition,recurrent stones and/or cholangitis were associated with residual stones ( OR:3.059,95% CI:1.307 - 7.159),stricture ( OR:3.702,95% CI:1.567-8.745) and endoscopic therapy (OR:4.841,95% CI:1.946 - 12.043) (P < 0.05).Conclusions Stricture,stone in both lobes and endoscope therapy were independent risk factors for residual stones; Residual stones,bile duct stricture and endoscope therapy were independent risk factors for recurrent stones and/or cholangitis.
2.The value of serum soluble cluster differentiation CD163 to the prognosis of patients with stroke associated pneumonia
Weidong ZHU ; Yunhua ZHANG ; Yifei WANG ; Qiang FANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2016;23(5):490-494
ObjectiveTo estimate the value of serum soluble scavenger receptor CD163 (sCD163) to the prognosis of patients with stroke associated pneumonia (SAP).Methods A prospective study was conducted. The clinically suspected SAP patients admitted to Department of Intensive Care Unit (ICU) and Emergency ICU (EICU) in Zhejiang Province Zhuji City Chinese Medicine Hospital from February 2014 to January 2015 were all enrolled. According to clinical pneumonia severity index (PSI), they were divided into SAP group and non SAP group according to the presence or absence of SAP, the patients of SAP group were subdivided into mild SAP group (PSI grade Ⅰ-Ⅲgrade) and severe SAP groups (PSI grade Ⅳ-Ⅴ grade) and according to the 28-day prognosis, the patients were subdivided into hospitalized death group and survival group. The clinical data were collected, including gender, age, history of stroke presence or absence, present stroke pattern, National Institutes of Health Stroke Scale score (NIHSS) on the day of stroke suspect diagnosis, Glasgow coma scale (GCS) score, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score, clinical pulmonary infection score (CPIS), PSI score, body temperature, result of chest X-ray film, oxygenation index (PaO2/FiO2), result of sputum culture, serum levels of white blood cell (WBC) and C-reactive protein (CRP) in a period of 7 days after the suspected diagnosis, PCT, sCDl63 levels on days 1, 3, 5, 7 days, length of stay in ICU, the total time of hospitalization and 28-day survival situation, etc. The ability of each index to evaluate the prognosis of SAP was analyzed by the receiver operating characteristic curve (ROC curve). The risk factors influencing the prognosis of SAP patients were analyzed by multivariate logistic regression analysis.Results ① Seventy-eight patients were finally enrolled in the study, 44 patients were diagnosed as SAP, 34 were non SAP. In 44 patients with SAP, there were 28 cases of severe SAP and 16 cases of mild SAP. On the first day of the suspected diagnosis, the NIHSS score [13 (7, 22) vs. 8 (4, 17), the CPIS score [6 (4, 9) vs. 4 (3, 5), sCD163 [mg/L: 0.80 (0.59, 1.32) vs. 0.33 (0.22, 0.46)], CRP [mg/L: 84.2 (50.8, 114.9) vs. 51.4 (26.2, 79.9)] and 28-day mortality [38.6% (17/44) vs. 11.8% (4/34)] in SAP group were significantly higher than those in non SAP group (allP < 0.05), and other basic indexes between the two groups were all of no statistical significant differences (allP > 0.05). ② The levels of sCD163 reached the peak value on the third day after the suspected diagnosis among SAP group and non SAP group, mild SAP group and severe SAP group, survival group and death group and then began to fall; the levels of sCD163, WBC, CRP, PCT within 7 days in SAP, severe SAP and death groups were higher than those in non SAP, mild SAP and survival groups (allP < 0.05). ③ROC curve analysis indicated: sCDl63 showed a better capacity for evaluating the 28-day prognosis of SAP [ROC curve (AUC) =0.673, 95% confidence interval (95%CI) = 0.515-0.807, sensitivity and specificity were 41.2% and 96.3% respectively and the cut-off was 2.65 mg/L]. However, the levels of other inflammatory indexes and scores on the first day after the suspected diagnosis had no value for early prognosis of SAP. The multiple logistic regression analysis showed that the level of sCD163 on the first day after the suspected diagnosis was the independent risk factor of death in hospital of SAP patients [dominance ratio (OR = 1.27, 95%CI = 1.06-1.52,P < 0.05]. Age (OR = 1.04, 95%CI = 1.01-1.06,P = 0.015), NIHSS score (OR = 2.86, 95%CI = 1.64-4.92,P = 0.010), CPIS score (OR = 1.52,95%CI = 1.28-1.90,P < 0.001) and APACHEⅡ score (OR = 2.06, 95%CI = 1.53-3.07,P < 0.001) were also the risk factors of influencing the death of patients with SAP.Conclusions Early sCD163 level is an independent risk factor in predicting the 28-day mortality of patients with SAP, and it has a certain value for the prognosis of SAP.
3.Expression and significance of signal transducers and activators of transcription 3 pathway genes in cholangiocarcinoma
Zhaojun HUANG ; Qiang HUANG ; Chenhai LIU ; Fang XIE ; Chenglin ZHU
Chinese Journal of Digestive Surgery 2014;13(12):971-976
Objective To investigate the expression of STAT3 signaling pathway genes including Survivin and COX-2 in cholangiocarcinoma,as well as the relationship between expression of these genes and prognosis of patients with cholangiocarcinoma.Methods The tumor and normal tissue samples were respectively collected from 43 patients with cholangiocarcinoma and 12 patients with intra-and extrahepatic bile duct stones or hepatic duct injury in the Affiliated Provincial Hospital of Anhui Medical University from September 2007 to July 2012.The expression of STAT3,phosphorylated-STAT3 (p-STAT3),Survivin and COX-2 were examined using immunohistochemistry,and the relationship between the expression of these genes and the clinical pathological features and prognosis of patients with cholangiocarcinoma was analyzed.Patients were followed-up through outpatient examination and telephone interview until March 2014.Categorical data were analyzed using the chi-square test.Correlation analysis was done by Spearman's method.The survival curve was generated using the Kaplan-Meier method,and the survival analysis was conducted using the log-rank test.Results The positive expression rates of STAT3,p-STAT3,Survivin and COX-2 in the tumor samples were 69.8% (30/43),65.1% (28/43),72.1% (31/43),79.1% (34/43),respectively,which were compared with 41.7% (5/12),8.3% (1/12),16.7% (2/12) and 41.7% (5/12) in the normal tissue samples,showing a significant difference for the last 3 indexes (x2=12.136,9.811,4.679,P < 0.05).Overexpression of p-STAT3,Survivin and COX-2 protein was correlated with lymph node metastasis (x2 =14.700,5.959,4.075,P < 0.05).Overexpression of p-STAT3 was also related to neural invasion (x2=10.384,P < 0.05).Expression of Survival and COX-2 protein was not associated with lymph invasion (x2=2.718,3.024,P > 0.05).Expression of p-STAT3,Survivin and COX-2 was however not associated with gender,age and tumor location,differentiation and diameter (x2=0.148,0.720,1.835,1.040,0.236 ; 0.001,0.009,0.029,1.863,0.197 ; 0.433,0.686,0.002,2.974,0.029,P > 0.05).Expression of Survivin and COX-2 protein was positively correlated to p-STAT3 protein (r =0.524,0.583,P < 0.05).All the 43 patients were followed up for 6-60 months.Among the 17 patients with hilar cholangiocarcinoma,the median survival time was 7,9,9 months for patients with positive expression of p-STAT3,Survivin and COX-2 protein,compared with 18,11 and 11 months for patients with negative expression of these proteins.The survival rates of the patients with positive and negative expression of p-STAT3 protein were 33.3% and 68.6%,respectively,with a statistical significance for p-STAT3 protein (x2=12.916,P < 0.05).Of the remaining 26 patients with common bile duct carcinoma,the median survival time was 9,10 and 9 months for patients with positive expression of p-STAT3,Survivin and COX-2 protein,compared with 20,20 and 20 months for patients with negative expression of these proteins.The survival rates of the patients with positive expression of p-STAT3,Survivin and COX-2 protein were 20.8%,9.4% and 8.5%,which were lower than 37.5%,37.5% and 50.0% of patients with the negative expression of these proteins,with a statically significance for all the 3 proteins (x2=12.787,6.245,11.161,P < 0.05).Conclusions The p-STAT3,Survivin,COX-2 proteins are highly expressed in the cholangiocarcinoma and the expression levels of these proteins are positively correlated.The survivin and COX-2 may be the downstream genes of STAT signaling pathway,which are involved in the progression and prognosis of cholangiocarcinoma.
4.Effects of suppressing the expression of miRNA-21 on the apoptosis and invasion abilities of cholangiocarcinoma cells and its target gene
Hao YOU ; Qiang HUANG ; Chenhai LIU ; Fang XIE ; Kai ZHU
Chinese Journal of Hepatobiliary Surgery 2011;17(9):760-763
ObjectiveTo discuss the effects of apoptosis and invasion of RBE cells caused by miRNA 21 suppression and further investigate the potential role of miRNA-21 plays on target mRNA regulation. MethodsThe RNAi technology was employed to suppress the expression of RBE cells.The changes in RECK mRNA and protein expressions were detected by RT-PCR and Western blotting respectively. Changes occurred in apoptosis was closely monitored by flow cytometry (FCM). The invasion of RBE cells was analyzed in vitro by invasion assay (transwell). ResultsThe expression of miRNA-21 was clearly suppressed while the RECK mRNA and protein were over-expressed. The rate of apoptosis was significantly accelerated and there was a dramatic decrease in RBE cells' ability to invade after miRNA-21 knockdown. ConclusionThrough miRNA-21 suppression, the rate of apoptosis of RBE cells was accelerated whereas their invasion ability was greatly reduced. RECK was found to be the target gene of miRNA-21 which participates in the regulation process of regulation.
5.Application of two dimensional ultrasound in the fine needle aspiration biopsy of thyroid nodules
Ying ZHANG ; Hongzhi MA ; Qiang ZHU ; Honggang LIU ; Jugao FANG
International Journal of Surgery 2012;39(6):380-383
Objective In this paper,we selected the ultrasound features of the malignant thyroid nodules to evaluate the preoperative diagnostic value of ultrasound-guided fine needle aspiration biopsy ( US-FNAB )in the preoperative evaluation of thyroid nodules.Methods Thirty-nine patients with thyroid nodules managed in the department of otolaryngology-head and neck surgery,Beijing Tongren Hospital,Captial Medical University from Dec.2010 to Aug.2011 were studied.All of them were undertaken the ultrasonic examination,ultrasound-guided fine needle aspiration biopsy and surgical treatment subsequently.The ultrasonic features,fine needle aspiration biopsy results and postoperative pathological results were analyzed to draw out the characteristics of sonographic features of thyroid malignant nodules.Results The sensitivity,specificity,misdiagnosis and missed diagnosis of fine needle aspiration biopsy were 80.0%,89.4%,11.0% and 20.0% respectively.Solid nodules,shape/dimension ratio ≥ 1,calcifications < 2 mm,irregular shape,blurred margins,type Ⅲ vascularity were correlated significantly with the malignant nodules of thyroid.The specificity of shape to dimension ratio and outline of the nodules were relatively higher than others (90.5%and 85.7% respectively).But there were no single ultrasound feature that could carry a high sensitivity and positive predictive value in the diagnosis of thyroid nodules.Conclusions Two ultrasonographic characteristics could be used together as a select indicator to screen malignant nodules which can reduce the chance of undertaking the fine needle aspiration biopsy in thyroid nodules.
6.Protective effect of melatonin against spinal cord injury from seawater immersion in rabbits
Tao WANG ; Chengru WU ; Haifeng WANG ; Jie ZHU ; Gewei LI ; He HONG ; Qiang YANG ; Jian FANG
Medical Journal of Chinese People's Liberation Army 2017;42(2):134-139
Objective To observe the protective effects of melatonin against spinal cord injury from seawater immersion in rabbits.Methods The 120 mature and health New Zealand White rabbits,weight range from 2.6 to 2.9kg,were randomly divided into four groups (30 each):control group,ethanol group,melatonin group (100mg/kg),methylPrednisolone group (30mg/kg).The rabbit model of spinal cord injury were built by modified Allen's method taking the 10th thoracic vertebra as a center,seawater immersion for 60 minutes,and then by grouping to give the appropriate treatment.After each group was given the corresponding treatment,six rabbits in each group were randomly selected at 1,6,12,24 and 48 hours five different time points.The neurological function scores of the rabbits were evaluated by Tarlov method,the spinal cord ofT9 to T.which were obtained from all the groups were used for further study,including immunohistochemical detection of apoptosis proteins:Bax,Bcl-2,neurofilament protein 200 (NF200) and in situ end labeling (TUNEL) method to detect spinal neuronal cell apoptosis.Results Within each observation time point,the Tarlov score was higher in melatonin group and methylprednisolone group compared with control group and ethanol group (P<0.05),there was no significant difference between melatonin group and methylprednisolone group (P>0.05).The expressions of Bcl-2 and NF200 were significantly higher in melatonin group and methylprednisolone group compared with control group and ethanol group,while Bax expression was significantly lower (P<0.05).There were no significant difference between melatonin group and methylprednisolone group in the expression of three proteins (P>0.05).The TUNEL-positive apoptotic cells were fewer in melatonin group and methylprednisolone group compared with control group and ethanol group (P<0.05),and there was no significant difference between melatonin group and methylprednisolone group (P>0.05).Conclusion Melatonin has protective effect against spinal cord injury from seawater immersion in rabbits,no difference in efficacy exists compare with methylprednisolone.
7.Correlation between sex hormone level in serum and expressed prostatic secretion with erectile function in patients with type Ⅲ prostatitis
Xiaoma ZHANG ; Jun FANG ; Jizhong CHEN ; Qiang GONG ; Youyun ZHOU ; Junjie QIAN ; Jinhai ZHU
Chongqing Medicine 2015;(11):1490-1492,1495
Objective To explore the correlation between the levels of estradiol E2 and testosterone T in serum and expressed prostatic secretion(EPS) with the erectile function in the patients with type Ⅲ prostatitis(CP/CPPS) .Methods The E2 and T lev‐els in serum and EPS from 64 cases of CP/CPPS ,including 35 cases of type Ⅲ A and 29 cases of Ⅲ B ,and 20 individuals of physical examination were detected by using the radioimmunoassay .All cases were evaluated by the scores of NIH‐CPSI and the Internation‐al Index of Erectile Function 5(IIEF‐5) .64 patients were grouped according to the IIEF‐5 scores ,the erectile dysfunction(ED) group(32 cases) and the non‐ED group(32 cases) .Results The mean E2/T levels in serum and EPS of the Ⅲ A group and the Ⅲ B group were higher than those in the control group ,the difference had statistical significance(P<0 .05) .20 cases(57 .14% ) of ED were found in the Ⅲ A group ,which were more than 12 cases(41 .38% ) of ED in the Ⅲ B group ,but there was no statistically signifi‐cant difference (>0 .05 .There was a positive correlation between the IIEF‐5 score and the T level in serum and EPS in the CP/CPPS group(r=0 .218 ,r=0 .231 ,P<0 .05) .There was a negative correlation between the IIEF‐5 score and the E2/T level in ser‐um and EPS(r= -0 .189 ,r= -0 .652 ,P<0 .05) ,which had no correlation with the NIH‐CPSI score(P>0 .05) .The serum T level in the ED group was (6 .32 ± 1 .86)ng/mL ,which was lower than(7 .89 ± 2 .92)ng/mL in the non‐ED group and (8 .41 ± 2 .02)ng/mL in the control group ;the .E2/T level in EPS in the ED group was (55 .02 ± 29 .26) ,which was higher than (14 .06 ± 9 .36) in the non‐ED group and (16 .45 ± 13 .76) in the control group ,the differences among them were statistically significant (P<0 .05) .Con‐clusion The imbalance degree of hormone estradiol and testosterone in serum and EPS is related with erectile function in the pa‐tients with CP/CPPS .
8.Dynamic predictive modeling of extraction process for red ginseng using near-infrared spectroscopy.
Jie-Qiang ZHU ; Wan-Fang PAN ; Yi ZHONG ; Xiao-Hui FAN ; Li-Yuan KANG ; Zheng LI
China Journal of Chinese Materia Medica 2014;39(14):2660-2664
It is the objective of this study to develop dynamic predictive model for the extraction process of red Ginseng using NIR spectroscopy. NIR spectroscopy was collected online and PLSR models were developed for total quantity of ginsenosides. The performance of NIR prediction model achieved R, RMSEC, RMSEP of 0.996 09, 0.018 9, 0.016 8, respectively. A first order dynamic mass transfer model was combined with NIR prediction of the quality indicator to predict the trajectory of the extraction process based upon the initial 3 or 4 data points. The results showed good agreement with actual measurements indicating reasonable accuracy of the predictive model. It could potentially be used for advanced predictive control of the extraction process.
Chemical Fractionation
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methods
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Ginsenosides
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chemistry
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isolation & purification
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Models, Theoretical
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Panax
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chemistry
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Spectroscopy, Near-Infrared
9.Expression level and significance of NGF, TGF-β1, estradiol, testosterone in serum and expressed prostatic secretion of patients with category Ⅲ prostatitis
Xiaoma ZHANG ; Jun FANG ; Jizhong CHEN ; Qiang GONG ; Youyun ZHOU ; Junjie QIAN ; Jinhai ZHU
Chinese Journal of Urology 2014;(7):518-523
Objective To investigate the level and clinical significance of nerve growth factor ( NGF) , transforming growth factor ( TGF )-β1 , estradiol ( E2 ) and testosterone ( T ) in serum and ex-pressed prostatic secretion (EPS) of patients with category Ⅲprostatitis. Methods From August 2011 to January 2012, 64 patients with (chronic prostatitis/chronic pelvic pain syndrome , CP/CPPS) and 20 health people were enrolled in this study.In CP/CPPS group, the age of patients ranged from 18 to 56 years, mean (36.6±9.3) years.The history of CP/CPPS ranged from 3 months to 6 years, mean 2 years.All patients were asked to complete NIH-CPSI questionnaires with CP/CPPS, including group ⅢA 35 cases and groupⅢB 29 cases.The age of healthy controls ranged from 25 to 41 years.The average healthy control age was (33.1±3.9) years.EPS and serum samples from CP/CPPS and control group were collected and frozen . NGF, TGF-β1 , E2 and T level in EPS and serum were measured by ELISA and radioimmunoassay and com -pared in each group. Results The mean E2, E2/T, TGF-β1 level in serum of patients with CP/CPPS were (175.7±82.4) pmol/L, (7.9±6.7), (2 216.2±581.6) ng/L, which were higher than that in healthy controls, (131.7±49.4) pmol/L, (4.6±2.4), (1 599.8±469.5) ng/L.The mean T level in CP/CPPS pa-tients′serum was (24.7±8.9) nmol/L, which was lower than that in controls (29.2±7.0) nmol/L.The E2/T (34.5±29.8), TGF-β1(6 859.3±5 229.4 ng/L), NGF (467.0±164.3 ng/L) levels in EPS of CP/CPPS patients were higher than that in controls (16.5±13.8), (1 774.1±1 304.3) ng/L, (310.8±106.6) ng/L. The TGF-β1 level in EPS of CP/CPPS patients showed the positive correlation ship with urination symptom score (6.1±2.4) (r=0.641, P<0.05).The NGF level in EPS of CP/CPPS patients also showed the positive correlation ship with pain score (7.6±2.6) (r=0.497, P<0.05).E2/T,TGF-β1 levels in serum and E2/T, TGF-β1,NGF levels in EPS of group ⅢA were (7.1±4.6), (2131.5±412.0)ng/L and (31.5±22.3), (7 667.1±5 652.4)ng/L, (440.6±134.3)ng/L, which were significantly higher than those in healthy con-trol (P<0.05).E2/T, TGF-β1 levels in serum and E2/T, TGF-β1, NGF levels in EPS of group ⅢB were (8.9±8.5), (2 340.5±728.2) ng/L and (38.2±37.1), (5 884.4±4 574.3) ng/L, (498.9±192.1) ng/L, which were also higher than those in healthy control ( P<0.05) . Conclusions Hormonal imbalance in es-tradiol and testosterone with TGF-β1 , NGF higher levels in EPS is closely related with pathogenesis and clin-ical symptom of category III chronic nonbacterial prostatitis .
10.Effects of extracellular signal-regulated kinase/protein kinase B signal pathway in cholangiocarcinoma cells invasion and migration promoted by microRNA-21
Zhiyuan JIN ; Qiang HUANG ; Chenhai LIU ; Zhen LIU ; Chenglin ZHU ; Fang XIE ; Chao WANG
Chinese Journal of Digestive Surgery 2016;15(9):918-924
Objective To observe the effects of extracellular signal-regulated kinase (ERK) 1/2 and protein kinase B (Akt) signal pathway in cholangiocarcinoma cells invasion and migration promoted by microRNA-21 (miR-21).Methods The experimental study was adopeted.QBC939 cholangiocarcinoma cells were cultured in vitro,through constructing and synthesizing unrelated sequence,miR-21 mimics and miR-21 inhibitor which were transfected into cells,and these cells were allocated into 4 groups,including growing naturally cells in the cell group,cells transfected by unrelated sequence in the 21-NC group,cells transfected by miR-21 mimics in the 21-M group and cells transfected by miR-21 inhibitor in the 21-Ⅰ group.Besides,cells in the 21-M group were allocated again into the 2 groups,20 μmol/L LY294002 and 10tμmol/L U0126 were respectively added in order to dispose 48 hours for follow-up experiments.Indicatiors of the test:(1) real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-21 in each group of cholangiocarcinoma cells.(2) Werstern blot was performed to detect the relative expressions of PTEN,ERK and Akt proteins in each group of cholangiocarcinoma cells.(3) Scarification assay was executed to test the migration of each group of cholangiocarcinoma cells.Transwell experiment was conducted to examine the migration and invasion of each group of cholangiocarcinoma cells.The measurement data with normal distribution were presented by x-s.The means of the 2 groups were compared by the t test.The means among groups were compared by the ANOVA,and pairwise comparison was analyzed by the Bonferroni test.The repeated measurement data were analyzed by the repeated measures ANOVA.Results (1) The relative expression of miR-21 in the cell group,21-NC group,21-M group and 21-Ⅰ group were 1.010 ±0.010,0.980 ± 0.050,4.900 ± 0.350 and 0.260 ± 0.010,respectively,with a statistically significant difference among the 4 groups (F =78.23,P < 0.05),with no statistically significant difference between the 21-NC group and cell group (P >0.05).There was increased expression between the 21-M group and cell group,decreased expression between the 21-Ⅰ group and cell group and significant difference between 21-M group or 21-Ⅰ group and cell group (P < 0.05).(2) The relative expressions of PTEN,ERK,p-ERK,Akt and p-Akt proteins in the cell group,21-NC group,21-M group and 21-Ⅰ group were 0.360 ± 0.020,0.400 ± 0.030,0.140 ± 0.010,0.680 ± 0.110 and 0.045 ± 0.126,0.470 ± 0.140,0.460 ± 0.060,0.440 ± 0.110 and 0.310 ± 0.020,0.380 ± 0.040,0.590 ± 0.060,0.160 ±0.010 and 0.400 ±0.010,0.390 ±0.080,0.410 ±0.090,0.380 ±0.070 and 0.440 ±0.110,0.510 ± 0.120,0.980 ± 0.150,0.190 ±0.010,respectively,showing statistically significant differences among the4 groups (F =10.23,12.78,18.11,P < 0.05).There was no significant difference in the relative expressions of PTEN,ERK,p-ERK,Akt and p-Akt proteins between the cell group and 21-NC group (P >0.05).Compared with cell group,there was decreased PTEN expression and increased p-ERK and p-Akt expressions in the 21-M group,showing statistically significant differences (P < 0.05).Compared with cell group,there was increased PTEN expression and decreased p-ERK and p-Akt expressions in the 21-Ⅰ group,showing statistically significant differences (P < 0.05).(3) The change of migration rate of cells from 6 hours to 48 hours were from 12.0% ± 3.0% to 23.0% ± 5.0% in the cell group,from 21.0% ± 4.0% to 43.0% ± 7.0% in the 21-M group,from 6.0% ±1.0% to 18.0% ±4.0% in the miR-21 + LY294002 group and from 9.0% ±2.0% to 26.0% ± 6.0% in the miR-21 + U0126 group,respectively.The migration rate of cells in the 21-M group at each time point was higher than that in the cell group (F =16.23,P <0.05).The migration rate of cells in the miR-21 + LY294002 group and miR-21 + U0126 group were lower than that in the 21-M group (F =25.21,P < 0.05),and there was the interaction effects between the change of migration rate of cells of the 3 groups and time,with a statistically significant difference (F =35.31,P < 0.05).(4) The numbers of migration cells in the cell group,21-M group,miR-21 + LY294002 group and miR-21 + U0126 group were 198 ± 32,248 ± 39,187 ±23 and 174 ± 28,respectively,with a statistically significant difference among the 4 groups (F =8.48,P < 0.05) and between the 21-M group and cell group (t =4.13,P <0.05).Compared with the 21-M group,the numbers of migration cells in the miR-21 + LY294002 group and miR-21 + U0126 group were decreased (F =21.98,P <0.05).The numbers of invasion cells in the cell group,21-M group,miR-21 + LY294002 group and miR-21 + U0126 group were 102 ± 22,211 ± 36,55 ± 9 and 67 ± 13,respectively,showing a statistically significant difference among the 4 groups (F =11.32,P < 0.05) and between the 21-M group and cell group (t =6.67,P < 0.05).Compared with the 21-M group,the numbers of invasion cells in the miR-21 + LY294002 group and miR-21 + U0126 group were decreased (F =36.23,P < 0.05).Conclusion ERK and Akt signal pathway participate in the cholangiocarcinoma cells invasion and migration promoted by miR-21,PTEN could mediate the process of promoting cholangiocarcinoma cells invasion and migration through ERK and Akt signal pathway promoted by miR-21.