1.Surgical treatment of primary liver cancer:a report of 10 966 cases
Yongxiang XIA ; Feng ZHANG ; Xiangcheng LI ; Lianbao KONG ; Hui ZHANG ; Donghua LI ; Feng CHENG ; Liyong PU ; Chuanyong ZHANG ; Xiaofeng QIAN ; Ping WANG ; Ke WANG ; Zhengshan WU ; Ling LYU ; Jianhua RAO ; Xiaofeng WU ; Aihua YAO ; Wenyu SHAO ; Ye FAN ; Wei YOU ; Xinzheng DAI ; Jianjie QIN ; Menyun LI ; Qin ZHU ; Xuehao WANG
Chinese Journal of Surgery 2021;59(1):6-17
Objective:To summarize the experience of surgical treatment of primary liver cancer.Methods:The clinical data of 10 966 surgically managed cases with primary liver cancer, from January 1986 to December 2019 at Hepatobiliary Center, the First Affiliated Hospital of Nanjing Medical University, were retrospectively analyzed. The life table method was used to calculate the survival rate and postoperative recurrence rate. Log‐rank test was used to compare the survival process of different groups, and the Cox regression model was used for multivariate analysis. In addition, 2 884 cases of hepatocellular carcinoma(HCC) with more detailed follow‐up data from 2009 to 2019 were selected for survival analysis. Among 2 549 patients treated with hepatectomy, there were 2 107 males and 442 females, with an age of (56.6±11.1) years (range: 20 to 86 years). Among 335 patients treated with liver transplantation, there were 292 males and 43 females, with an age of (51.0±9.7) years (range: 21 to 73 years). The outcomes of hepatectomy versus liver transplantation, anatomic versus non-anatomic hepatectomy were compared, respectively.Results:Of the 10 966 patients with primary liver cancer, 10 331 patients underwent hepatectomy and 635 patients underwent liver transplantation. Patients with liver resection were categorized into three groups: 1986-1995(712 cases), 1996-2008(3 988 cases), 2009?2019(5 631 cases). The 5‐year overall survival rate was 32.9% in the first group(1986-1995). The 5‐year overall survival rate of resected primary liver cancer was 51.7% in the third group(2009‐2019), among which the 5‐year overal survival rates of hepatocellular carcinoma, intrahepatic cholangiocarcinoma and mixed liver cancer were 57.4%, 26.6% and 50.6%, respectively. Further analysis was performed on 2 549 HCC patients with primary hepatectomy. The 1‐, 3‐, 5‐, and 10‐year overall survival rates were 88.1%, 71.9%, 60.0%, and 41.0%, respectively, and the perioperative mortality rate was 1.0%. Two hundred and forty‐seven HCC patients underwent primary liver transplantation, with 1‐, 3‐, 5‐, and 10‐year overall survival rates of 84.0%, 64.8%, 61.9%, and 57.6%, respectively. Eighty‐eight HCC patients underwent salvage liver transplantation, with the 1‐, 3‐, 5‐, and 10‐year overall survival rates of 86.8%, 65.2%, 52.5%, and 52.5%, respectively. There was no significant difference in survival rates between the two groups with liver transplantation ( P>0.05). Comparing the overall survival rates and recurrence rates of primary hepatectomy (2 549 cases) with primary liver transplantation (247 cases), the 1‐, 3‐, 5‐, and 10‐year overall survival rates in patients within Milan criteria treated with hepatectomy and transplantation were 96.3%, 87.1%, 76.9%, 54.7%, and 95.4%, 79.4%, 77.4%, 71.7%, respectively ( P=0.754). The 1‐, 3‐, 5‐year recurrence rates were 16.3%, 35.9%, 47.6% and 8.1%, 11.7%, 13.9%, respectively( P<0.01). The 1‐, 3‐, 5‐, 10‐year overall survival rates in patients with no large vessels invasion beyond the Milan criteria treated with liver resection and transplantation were 87.2%, 65.9%, 53.0%, 33.0% and 87.6%, 71.8%, 71.8%, 69.3%, respectively( P=0.003); the 1‐, 3‐, 5‐year recurrence rate were 39.2%, 57.8%, 69.7% and 29.7%, 36.7%, 36.7%, respectively ( P<0.01). The 1‐, 3‐, 5‐, and 10‐year overall survival rates in patients with large vessels invasion treated with liver resection and transplantation were 62.1%, 36.1%, 22.2%, 15.0% and 62.9%, 31.8%,19.9%, 0, respectively ( P=0.387); the 1‐, 3‐, 5‐year recurrence rates were 61.5%, 74.7%, 80.8% and 59.7%, 82.9%, 87.2%, respectively( P=0.909). Independent prognostic factors for both overall survival and recurrence‐free survival rates of HCC patients treated with liver resection included gender, neoadjuvant therapy, symptoms, AST, intraoperative or postoperative blood transfusion, tumor number, tumor size, cirrhosis, macrovascular invasion, microvascular invasion, and pathological differentiation. Propensity score matching analysis of 443 pairs further showed that there was no significant difference in overall survival rate between anatomical liver resection and non‐anatomical liver resection( P=0.895), but the recurrence rate of non‐anatomical liver resection was higher than that of anatomical liver resection( P=0.035). Conclusions:In the past decade, the overall survival rate of HCC undergoing surgical treatment is significantly higher than before. For HCC patients with good liver function reservation, surgical resection can be performed first, and salvage liver transplantation can be performed after recurrence. The effect of salvage liver transplantation is comparable to that of primary liver transplantation. As for the choice of liver resection approaches, non‐anatomical resection can reserve more liver tissue and can be selected as long as the negative margin is guaranteed.
2.Surgical treatment of primary liver cancer:a report of 10 966 cases
Yongxiang XIA ; Feng ZHANG ; Xiangcheng LI ; Lianbao KONG ; Hui ZHANG ; Donghua LI ; Feng CHENG ; Liyong PU ; Chuanyong ZHANG ; Xiaofeng QIAN ; Ping WANG ; Ke WANG ; Zhengshan WU ; Ling LYU ; Jianhua RAO ; Xiaofeng WU ; Aihua YAO ; Wenyu SHAO ; Ye FAN ; Wei YOU ; Xinzheng DAI ; Jianjie QIN ; Menyun LI ; Qin ZHU ; Xuehao WANG
Chinese Journal of Surgery 2021;59(1):6-17
Objective:To summarize the experience of surgical treatment of primary liver cancer.Methods:The clinical data of 10 966 surgically managed cases with primary liver cancer, from January 1986 to December 2019 at Hepatobiliary Center, the First Affiliated Hospital of Nanjing Medical University, were retrospectively analyzed. The life table method was used to calculate the survival rate and postoperative recurrence rate. Log‐rank test was used to compare the survival process of different groups, and the Cox regression model was used for multivariate analysis. In addition, 2 884 cases of hepatocellular carcinoma(HCC) with more detailed follow‐up data from 2009 to 2019 were selected for survival analysis. Among 2 549 patients treated with hepatectomy, there were 2 107 males and 442 females, with an age of (56.6±11.1) years (range: 20 to 86 years). Among 335 patients treated with liver transplantation, there were 292 males and 43 females, with an age of (51.0±9.7) years (range: 21 to 73 years). The outcomes of hepatectomy versus liver transplantation, anatomic versus non-anatomic hepatectomy were compared, respectively.Results:Of the 10 966 patients with primary liver cancer, 10 331 patients underwent hepatectomy and 635 patients underwent liver transplantation. Patients with liver resection were categorized into three groups: 1986-1995(712 cases), 1996-2008(3 988 cases), 2009?2019(5 631 cases). The 5‐year overall survival rate was 32.9% in the first group(1986-1995). The 5‐year overall survival rate of resected primary liver cancer was 51.7% in the third group(2009‐2019), among which the 5‐year overal survival rates of hepatocellular carcinoma, intrahepatic cholangiocarcinoma and mixed liver cancer were 57.4%, 26.6% and 50.6%, respectively. Further analysis was performed on 2 549 HCC patients with primary hepatectomy. The 1‐, 3‐, 5‐, and 10‐year overall survival rates were 88.1%, 71.9%, 60.0%, and 41.0%, respectively, and the perioperative mortality rate was 1.0%. Two hundred and forty‐seven HCC patients underwent primary liver transplantation, with 1‐, 3‐, 5‐, and 10‐year overall survival rates of 84.0%, 64.8%, 61.9%, and 57.6%, respectively. Eighty‐eight HCC patients underwent salvage liver transplantation, with the 1‐, 3‐, 5‐, and 10‐year overall survival rates of 86.8%, 65.2%, 52.5%, and 52.5%, respectively. There was no significant difference in survival rates between the two groups with liver transplantation ( P>0.05). Comparing the overall survival rates and recurrence rates of primary hepatectomy (2 549 cases) with primary liver transplantation (247 cases), the 1‐, 3‐, 5‐, and 10‐year overall survival rates in patients within Milan criteria treated with hepatectomy and transplantation were 96.3%, 87.1%, 76.9%, 54.7%, and 95.4%, 79.4%, 77.4%, 71.7%, respectively ( P=0.754). The 1‐, 3‐, 5‐year recurrence rates were 16.3%, 35.9%, 47.6% and 8.1%, 11.7%, 13.9%, respectively( P<0.01). The 1‐, 3‐, 5‐, 10‐year overall survival rates in patients with no large vessels invasion beyond the Milan criteria treated with liver resection and transplantation were 87.2%, 65.9%, 53.0%, 33.0% and 87.6%, 71.8%, 71.8%, 69.3%, respectively( P=0.003); the 1‐, 3‐, 5‐year recurrence rate were 39.2%, 57.8%, 69.7% and 29.7%, 36.7%, 36.7%, respectively ( P<0.01). The 1‐, 3‐, 5‐, and 10‐year overall survival rates in patients with large vessels invasion treated with liver resection and transplantation were 62.1%, 36.1%, 22.2%, 15.0% and 62.9%, 31.8%,19.9%, 0, respectively ( P=0.387); the 1‐, 3‐, 5‐year recurrence rates were 61.5%, 74.7%, 80.8% and 59.7%, 82.9%, 87.2%, respectively( P=0.909). Independent prognostic factors for both overall survival and recurrence‐free survival rates of HCC patients treated with liver resection included gender, neoadjuvant therapy, symptoms, AST, intraoperative or postoperative blood transfusion, tumor number, tumor size, cirrhosis, macrovascular invasion, microvascular invasion, and pathological differentiation. Propensity score matching analysis of 443 pairs further showed that there was no significant difference in overall survival rate between anatomical liver resection and non‐anatomical liver resection( P=0.895), but the recurrence rate of non‐anatomical liver resection was higher than that of anatomical liver resection( P=0.035). Conclusions:In the past decade, the overall survival rate of HCC undergoing surgical treatment is significantly higher than before. For HCC patients with good liver function reservation, surgical resection can be performed first, and salvage liver transplantation can be performed after recurrence. The effect of salvage liver transplantation is comparable to that of primary liver transplantation. As for the choice of liver resection approaches, non‐anatomical resection can reserve more liver tissue and can be selected as long as the negative margin is guaranteed.
3.Effects of non-steroidal anti-inflammatory drugs administered at different time points on mesenteric traction syndrome following open radical gastrectomy in elderly patients
Zheng CHEN ; Donghua SHAO ; Zumin MAO ; Xiaodong MA
Chinese Journal of Geriatrics 2020;39(6):666-671
Objective:To investigate the effects of the non-steroidal anti-inflammatory drugs Parecoxib and Flurbiprofen administered at different time points on mesenteric traction syndrome(MTS).Methods:This was a prospective, randomized, controlled clinical trial.One hundred elderly patients scheduled for open radical gastrectomy under general anesthesia were randomly allocated to four groups: the control group, the P-Pre-MT group, the F-Pre-MT group, and the F-Post-MT group(n=25, each group). Parecoxib 40 mg and Flurbiprofen 50 mg were intravenously administered 30 min and 5 min before skin incision in the P-Pre-MTS group and the F-Pre-MTS group, respectively.Flurbiprofen 50 mg was infused at the moment of MTS in the F-Post-MTS group while the control group was intravenously injected with saline.Anesthesia induction and maintenance were performed with plasma target-controlled infusion of Propofol and Remifentanil.After the incision of the peritoneum.The incidence of MTS, the duration of hypotension, and the use of norepinephrine during MTS were recorded.Systolic blood pressure(SBP), heart rate(HR), and effect-site concentration of Remifentanil were monitored at MT(T 0), 10 min(T 10), 20 min(T 20), 30 min(T 30), 45 min(T 45), and 60 min(T 60)after MT in patients with MTS. Results:MTS was observed in 19 of 22 patients(86%), 19 of 23 patients(83%), 0 of 24 patients(0%)and 20 of 23 patients(87%)in the control, P-Pre-MT, F-Pre-MT and F-Post-MT groups, respectively.The incidence of MTS in the F-Pre-MT group was lower than that in the control group( χ2=35.313, P=0.000). The duration of hypotension and the use of norepinephrine in patients with MTS were less in the F-Post-MT group than in the control group( P=0.007 and 0.015). SBP and HR at different time points after MT had significant differences in patients with MTS in the control group( F=47.425 and 26.318, P=0.000 and 0.000), but did not differ in the F-Pre-MT group( F=2.140 and 1.013, P=0.066 and 0.413). SBP and the effect-site concentration of Remifentanil were lower and HR was higher in the control group than in the F-Pre-MT group at T 10and T 20after MT( P=0.000), and SBP was higher and HR was lower in the F-Post-MT group than in the control group C at T 20after MT( P=0.002 and 0.002). Conclusions:Flurbiprofen not only can prevent the occurrence of MTS, maintain blood pressure stability and heart rate after MT, but also can reduce the duration of hypotension and the amplitude of heart rate increase when MTS occurs in elderly patients undergoing open radical gastrectomy.Parecoxib has no effect on MTS.
4.Role of CaN/NFATc4 signaling pathway in ventilator-induced lung injury
Min LI ; Xiaotian LIU ; Yunbin XIE ; Yongfeng ZHENG ; Xiangzhi FANG ; Yan XIA ; Donghua SHAO
Chinese Journal of Anesthesiology 2019;39(6):761-764
Objective To evaluate the role of calcineurin ( CaN)/nuclear factor of activated T cell cytoplasmic 4 protein ( NFATc4) signaling pathway in inflammatory responses in lung tissues of rats with ventilator-induced lung injury ( VILI) . Methods Twenty-four clean-grade healthy male Wistar rats, aged 5-8 weeks, weighing 220-250 g, were divided into 3 groups ( n=8 each) using a random number table method: control C (group C), VILI group and cyclosporine A plus VILI group (group CsA+VILI). The animals were anesthetized with pentobarbital and tracheostomized. The rats were mechanically ventilated for 6 h with the tidal volume set at 40 ml/kg and respiratory rate at 40 breaths/min to establish the model of VI-LI. The rats kept spontaneous breathing in group C. CaN specific inhibitor cyclosporine A 10 mg/kg was in-traperitoneally injected at 1 h before ventilation in group CsA+VILI. Rats were sacrificed immediately after ventilation, lung tissues were obtained and stained with hematoxylin and eosin to evaluate lung injury, broncho-alveolar lavage fluid was collected for determination of tumor necrosis factor-alpha ( TNF-α) , inter-leukin-1beta ( IL-1β) and IL-6 concentrations by enzyme-linked immunosorbent assay, and the lungs were removed for determination of the wet to dry weight ratio ( W/D ratio) , expression of intercellular adhesion molecule-1 ( ICAM-1) and vascular cell adhesion molecule-1 ( VCAM-1) ( by real-time polymerase chain reaction) , and expression of calcineurin and NFATc4 in lung tissues ( using Western blot ) . Results Compared with group C, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-αin BALF were significantly increased, and the expression of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was up-regulated in group VILI ( P<0. 05) . Compared with group VILI, the W/D ratio, lung injury scores and concentrations of IL-1β, IL-6 and TNF-αin BALF were significantly decreased, and the expres-sion of CaN, NFATc4, ICAM-1 mRNA and VCAM-1 mRNA was down-regulated in group CsA+VILI ( P<0. 05) . Conclusion CaN/NFATc4 signaling pathway mediates inflammatory responses in lung tissues of rats with VILI.
5.Thoughts on the Construction of Humanistic Care System in Public Hospitals in Anhui Province
Tao XU ; Fuyong HU ; Li LI ; Qi YAN ; Shuang HU ; Hui FAN ; Donghua SHAO
Chinese Medical Ethics 2019;32(3):380-383,392
Humanistic care centered on the interests and needs of patients, is an important means to maintain the doctor-patient relationship and to build a harmonious society. However, there are still some problems in the major public hospitals, such as inadequate communication and coordination, complicated and time-consuming medical procedures, and so on. Based on this, this paper discussed the problems existing in public hospitals in Anhui Province, and put forward some measures, such as strengthening doctor-patient communication, caring for doctor-patient personnel, and constructing hospital culture, so as to perfect the humanistic care system.
6.Effect of dexmedetomidine on blood coagulation function following radical gastrectomy
Zheng CHEN ; Donghua SHAO ; Zumin MAO ; Leilei SHI ; Yongfeng ZHENG ; Dapeng ZHANG
The Journal of Clinical Anesthesiology 2017;33(11):1086-1090
Objective To explore the effect of dexmedetomidine on blood coagulation following radical gastrectomy.Methods ASA Ⅰ or Ⅱ patients aged 51-70 years weighing 53-75 kg scheduled for radical gastrectomy were randomly allocated to two groups:dexmedetomidine group (group D)and control group (group C).Dexmedetomidine 0.5 μg/kg was intravenously infused over 10 minutes before anesthesia induction,followed by a rate of 0.5 μg·kg-1 ·h-1 until peritoneal closure in group D and volume-matched normal saline was administrated in group C.Radical gastrectomy was performed under total intravenous anesthesia with propofol and remffentanil.A series of warming measures were implemented and artificial colloid and heparin flushing fluid were not used.Postoperative patient-controlled intravenous analgesia was performed to maintain visual analogue scale≤3.The blood samples were collected for TEG and standard coagulation monitoring before dexmedetomidine and saline administration and 3 h after surgery.Results The temperature and hematocrit in the postoperative period were significantly less than the preoperative period in two groups (P<0.01).In both groups,the activity of plasma antithrombin Ⅲ was significantly decreased and the concentration of plasma FDP was significantly increased in the postoperative period when compared with the preoperative period (P <0.01).In group D,the R time was significantly shortened and MA value was significantly increased in the postoperative period when compared with the preoperative period (P<0.05) and there were no significant differences in the K time and α angle between the preoperative and postoperative period.In group C,the R and K time were significantly shortened and the value for MA and α angle were significantly increased in the postoperative period compared with the preoperative period (P<0.01).The platelet counts,PT,APTT,and plasma fibrinogen concentration were comparable between the preoperative and postoperative period in both groups.The requirements of propofol and remifentanil in group D were significantly less than group C (P<0.05).In the preoperative period,the plasma antithrombin Ⅲ activity,FDP concentration,and the values for all TEG variables were similar in both groups.In the postoperative period,the value for MA and the concentration of plasma FDP in group D were less than that in group C and the value for R and the activity of plasma AT Ⅲ in group D were more than group C (P<0.05 or P<0.01) and there were no significant differences in the K time and α angle in both groups.There were no significant differences in the temperature,hematocrit,platelet counts,PT,APTT,and plasma fibrinogen concentration in the preoperative and postoperative periods between the two study groups.Conclusion Adjunctive dexmedetomidine in general anesthesia could inhibit the decrease of R time and raise of the value for MA,inhibit the decrease of plasma an tithrombin Ⅲ activity and raise of FDP concentration,which indicated that dexmedetomidine can improve blood coagulation state after radical gastrectomy.
7.Diagnostic Value of Postmortem CT Angiography in Coronary Atherosclerosis
Hui QIAN ; Yu SHAO ; Zhengdong LI ; Donghua ZOU ; Zhiqiang QIN ; Lei WAN ; Yijiu CHEN
Journal of Forensic Medicine 2017;33(2):109-113
Objective T o explore the application value of postm ortem com puted tom ography (C T ) an-giography on diagnosis of coronary atherosclerotic stenosis degree. Methods B ased on the previous ex-perim ental results, the postm ortem C T angiography device of hum an isolated heart w as im proved. D iffer-ent coronary atherosclerotic stenosis degree of sudden death cases w as selected. B efore the cardiac anatom y, hearts w ere rem oved out com pletely and C T angiography w as perform ed im m ediately. T he C T angiography results w ere com pared w ith histopathological findings. M eanw hile, the advantages and disad-vantages of the angiography device before and after im provem ent w ere com pared. Results T he im proved angiography device of isolated heart could get better im aging results. T he postm ortem C T angiography results had high consistency w ith the histopathological findings on diagnosis of coronary atherosclerotic stenosis degree. A nd the coronary artery lesions could be revealed m ore objectively and vividly by 3D reconstruction technology. H ow ever, C T angiography could only be used to exam ine the pathological changes of blood vessels, w hich m ight have som e lim itations on the diagnosis of cause of death. Con-clusion Postm ortem C T angiography can be used as an additional m ethod for the conventional autopsy in the cases of coronary atherosclerosis.
8.Optimal pressure for facemask ventilation during induction of general anesthesia in adult patients:real-time ultrasonographic measurement of antral cross-sectional area
Lihua HANG ; Shiyou WEI ; Zhenkai XU ; Weiwei SHU ; Yuanfeng CHEN ; Zheng CHEN ; Leilei SHI ; Donghua SHAO
Chinese Journal of Anesthesiology 2017;37(4):461-463
Objective To determine the optimal pressure for facemask ventilation during induction of general anesthesia by real-time ultrasonographic measurement of antral cross-sectional area (CSA) in adult patients.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 18-60 yr,with body mass index of 20-25 kg/m2,scheduled for elective operation under general anesthesia,were divided into 5 groups (n=12 each) using a random number table:P10 group,P13 group,P16 group,P19 group and P22 group.After induction of anesthesia,an oropharyngeal airway was inserted,and the patients were ventilated for a 2-min period in a pressure-controlled mode using the two-handed mask ventilation technique.The pressure for facemask ventilation was 10,13,16,19 and 22 cmH2O in P10,P13,P16,P19 and P22 groups,respectively.The antral CSA was measured using real-time ultrasonography before and after facemask ventilation.Respiratory parameters were recorded.Results Compared with group P1O,the number of patients in whom CSA<340 mm2 after facemask ventilation was significantly decreased in P16,P19 and P22 groups,and the number of patients in whom the tidal volume ≥ 6 ml/kg was increased in P13,P16,P19 and P22 groups (P< 0.01).The number of patients in whom optimnal pressure for facemask ventilation was achieved was 2,10,6,4 and 1 in P10,P13,P16,P19 and P22 groups,respectively,with the most cases in group P13 (P < 0.01).Conclusion The optimal pressure is 13 emH2O for facemask ventilation during induction of general anesthesia when determined by realtime ultrasonographic measurement of antral CSA,and it can ensure adequate oxygen supply and reduce gastric insufflation in adult patients.
9.Relationship between TDAG8 and endogenous neuron-protective mechanism against oxygen-glucose deprivation and restoration-induced apoptosis in rat neurons
Xiaodong MA ; Donghua SHAO ; Lihua HANG ; Weiwei SHU ; Xiulan HU ; Hong LUO
Chinese Journal of Anesthesiology 2016;36(9):1080-1084
Objective To investigate the relationship between T?cell death?associated gene 8 ( TD?AG8) and endogenous neuron?protective mechanism against oxygen?glucose deprivation and restoration ( OGD∕R)?induced apoptosis in rat neurons. Methods The primary cortical neurons obtained from fetal rats were seeded in 6?well plates at a density of 1×105 cells∕ml and divided into 5 groups using a random number table: control group ( group C, n=24 ) , group OGD∕R ( n=48 ) , TDAG8 agonist BTB09089 group (group BTB, n=24), TDAG8?siRNA group ( group siRNA, n=24), and blank vehicle group ( group V, n=24) . The medium was replaced with glucose?and serum?free Locke′s buffer, and the neu?rons were exposed to 95% N2?5% CO2 in an air?tight incubator at 37℃ for 60 min followed by routine cul?ture to establish the model of OGD∕R. In BTB, siRNA and V groups, 20 μmol∕L TDAG8 agonist
BTB09089, 200 pmol∕L TDAG8?siRNA, and 6 μl∕200 μl transfection reagent were added, respectively, at 24 h before oxygen?glucose restoration. At 6 h of oxygen?glucose restoration, the neuronal viability and a?mount of lactic dehydrogenase ( LDH) released were measured, and the expression of TDAG8 and caspase?3 mRNA in neurons was detected by fluorescent quantitative real?time polymerase chain reaction. In group OGD∕R, the expression of TDAG8 and caspase?3 was measured by Western blot at 0, 3, 6, 12 and 24 h of oxygen?glucose restoration. In C, OGD∕R, BTB, siRNA and V groups, the expression of TDAG8, caspase?3 and p?Akt was detected at 6 h of oxygen?glucose restoration. Results In group OGD∕R, the ex?pression of TDAG8 was gradually up?regulated after oxygen?glucose restoration, and the expression of caspase?3 peaked at 6 h of oxygen?glucose restoration. Compared with group C, the neuronal viability was significantly decreased, the amount of LDH released was significantly increased, and the expression of TD?AG8 and caspase?3 protein and mRNA and p?Akt was significantly up?regulated in OGD∕R, V and siRNA groups ( P<0?05) . Compared with group OGD∕R, the expression of TDAG8 protein and mRNA and p?Akt was significantly up?regulated, the expression of caspase?3 protein and mRNA was significantly down?regu?lated, the neuronal viability was significantly increased, and the amount of LDH released was significantly decreased in group BTB, the expression of TDAG8 protein and mRNA and p?Akt was significantly down?regulated, the expression of caspase?3 protein and mRNA was significantly up?regulated, the neuronal via?bility was significantly decreased, and the amount of LDH released was significantly increased in group siR?NA ( P<0?05) , and no significant change was found in the parameters mentioned above in group V ( P>0?05) . Conclusion TDAG8 is partially involved in the endogenous neuron?protective mechanism against OGD∕R?induced apoptosis in rat neurons, which may be related to activation of Akt signaling pathway.
10.Association between PNPLA3 polymorphisms (rs738409) and non-alcoholic fatty liver disease
Guowei LIANG ; Donghua SHAO ; Jie LIU ; Meilin HE
Chinese Journal of Laboratory Medicine 2016;(1):45-49
Objective To develop a novel method for genotyping rs 738409 ( C >G ) single nucleotide polymorphism ( SNP ) , and explore the association between the rs 738409 genotypes and non-alcoholic fatty liver disease ( NAFLD ) .Methods Method establishment and analysis of genetic susceptibility.The principle of amplification refractory mutation system ( ARMS) and combined a TaqMan fluorogenic probe as signal report were used , by monitoring the difference of cycle threshold (ΔCt=C allele-special primer Ct values minus G allele-special primer Ct values ) between the two PCR reactions in a real-time PCR, the method for rs738409 genotyping was established ( ARMS-TaqMan) .618 subjects ( men:401;women: 217 ) , in an annual health check-up program from January 2011 to December 2014 in Aerospace Center Hospital , were performed for rs738409 genotyping by the ARMS-TaqMan assay.Fatty liver was diagnosed based on abdominal ultrasonography .The chi-square test and multiple logistic analyses were used to analyze the relationship of rs 738409 genotypes and NAFLD.Results The ΔCt by ARMS-TaqMan for rs738409 genotyping were -13.1 ±1.4 of CC alleles ( 243 cases ) , 0.01 ±0.45 of CG alleles ( 282 cases), and 12.7 ±1.9 of GG alleles (93 cases), respectively.The GG alleles frequency of rs738409 were significantly higher in patients with NAFLD compared with subjects without NAFLD (21.5%vs 12.3%,χ2=8.677, P =0.003).In comparison to subjects with CC alleles, the OR (95% confidence interval) adjusted for age, gender, total cholesterol, triglyceride, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, fasting blood glucose and body mass index was 1.35 (0.91-2.00) in subjects with CG alleles and was 2.21 ( 1.32 -3.71 ) in subjects with GG alleles ( P =0.013 ) .Variant rs738409 genotypes were associated with significant increased trend in alanine aminotransferase ( ALT) level from CC alleles, CG alleles to GG alleles (F=8.980, P<0.001), and in aspartate aminotansferase (AST) between GG alleles and CC alleles (F=6.491, P<0.001).Conclusions The novel ARMS-TaqMan assay had the features of accuracy , one step and high-throughput for rs738409 genotyping.The G allele of rs738409 was a risk factor of NAFLD susceptibility and associated with higher level serum ALT and AST .

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