Objective To study the protective effects and mechanism of sacubitril(Sac)/valsartan(Val)on cardiomyocytes of rabbits with heart failure induced by doxorubicin(DOX).Methods Thirty New Zealand rabbits were selected to establish DOX-induced heart failure rabbit model.Twenty-five rabbits with successful modeling were randomly assigned to model group(DOX group,n=9),DOX+Val group(n=8),and DOX+Sac/Val group(n=8);and another 8 New Zealand rabbits were selected as blank group.The DOX+Val group was gavaged with 4.65 mg/kg Val suspension each time,the DOX+Sac/Val group was gavaged with 9.3 mg/kg Sac/Val suspension each time,and the blank group and DOX group were gavaged with equal volume of distilled water each time.Each group was gavaged twice a day for 8 weeks.After 8 weeks of administration,echocardiography was used to measure left ventricular end-diastolic diameter(LVDD),left ventricular end-systolic diameter(LVSD),left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS).The heart mass index(HMI)and left ventricular mass index(LVMI)were calculated.The pathological morphology and myocardial fibrosis of myocardial tissue were observed by hematoxylin-eosin(H-E)and Masson staining.The ultrastructure of cardiomyocytes was observed by transmission electron microscope.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was used to observe cardiomyocytes apoptosis and apoptosis rate was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP),high-sensitivity cardiac troponin I(Hs-cTNI),angiotensin Ⅱ(Ang Ⅱ),aldosterone(ALD),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP),and protein kinase G(PKG)in serum.Quantitative polymerase chain reaction(qPCR)was used to detect the expression of natriuretic peptide receptor A(NPR-A),cGMP-specific phosphodiesterase 5A(PDE5A[cGMP]),PKG,B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),and cysteine aspartate protease 3(caspase 3)mRNA in myocardial tissue.Western blotting was used to detect the expression of phosphorylated cAMP response element-binding protein(p-CREB),phosphorylated Bcl-2 related death promoting factor(p-Bad),Bcl-2,Bax,and caspase 3 proteins in myocardial tissue.Results Compared with the blank group,the LVDD and LVSD in the DOX group were increased(both P<0.01),the LVEF and LVFS were decreased(both P<0.01)and the HMI and LVMI were increased(both P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were increased(P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,p-CREB,Bax and caspase 3 were all increased(all P<0.01),while the expression of Bcl-2 was decreased(P<0.01),and the expression of p-Bad had no significant difference(P>0.05).Compared with the DOX group,the LVDD and LVSD of the DOX+Sac/Val group and DOX+Val group were decreased(all P<0.01),the LVEF and LVFS were increased(all P<0.01)and the HMI and the LVMI were decreased(all P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were decreased(all P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,Bax and caspase 3 were all decreased(all P<0.01),while the expression of Bcl-2 was increased(P<0.01);and the expression of p-CREB and p-Bad was increased in the DOX+Sac/Val group(both P<0.01),but there was no significant difference in the DOX+Val group(both P>0.05).Compared with the DOX+Val group,the DOX+Sac/Val group showed a decrease in all indicators except for LVEF,LVFS,NPR-A,ANP,BNP,cGMP,PDE5A(cGMP),PKG,p-CREB,p-Bad,and Bcl-2,which were all elevated(all P<0.05).Myocardial pathology and transmission electron microscopy showed that Sac/Val effectively protected cardiomyocytes,reduced cardiomyocytes apoptosis and myocardial fibrosis,and these effects were significantly better than those of Val.Conclusion Sac/Val can effectively reduce cardiomyocytes apoptosis,improve cardiac function and reduce myocardial fibrosis in rabbits with heart failure,and these effects are superior to Val.Its mechanism may be related to activating the NPR-A/cGMP/PKG signaling pathway and inhibiting renin-angiotensin-aldosterone system.

Objective To investigate the intervention effect of sacubitril/valsartan(Sac/Val)on doxorubicin-induced heart failure(HF)in rabbits and its regulative effect on the transforming growth factor β1(TGF-β1)/SMAD family member 3(Smad3)/connective tissue growth factor(CTGF)signaling pathway.Methods Thirty-eight male New Zealand rabbits were subjected,and 8 of them were randomly assigned into a control group.The other 30 rabbits were injected with doxorubicin to establish a rabbit HF model,and finally,there were 6 rabbits in a model group,7 in a valsartan group,and 7 in a Sac/Val group.After 8 weeks of intervention,echocardiography[left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD)],and myocardial histopathologic observation(HE staining,Masson staining)were performed,and collogen volume fraction(CVF)was calculated.The levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),soluble growth stimulation expressed gene 2(sST2),galectin-3(Gal-3)were detected.Activities of renin-angiotensin-aldosterone system(RAAS)and levels of atrial natriuretic peptide,B-type brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP)and protein kinase G(PKG)were measured.The expression of α-smooth muscle actin(α-SMA),typeⅠ collagen,TGF-β1,Smad3,recombinant SMAD family member 7(Smad7)and CTGF in the myocardial tissues were detected.Results Compared with the control group,the model group exhibited significantly lower LVEF and LVFS and decreased expression of Smad7,higher LVEDD and LVESD(P＜0.01).The CVF of each group was(7.15±0.82)％、(43.20±5.09)％、(29.53±4.05)％、(22.48±2.93)％.Valsartan and Sac/Val treatment resulted in obvious increases in LVEF and LVFS,up-regulation of Smad7,decreased in LVEDD,LVESD and CVF,reduced levels of NT-proBNP,sST2,Gal-3,AngⅡ,aldosterone,atrial natriuretic peptide,BNP,cGMP and PKG,and down-regulation of α-SMA,collagen I,TGF-β1,Smad3 and CTGF when compared with the model group(P＜0.01).Sac/Val treatment showed better effects in above indicators than simple valsartan treatment(P＜0.01).Conclusion Sac/Val can reduce myocardial fibrosis and improve cardiac function in doxorubicin-induced HF rabbits,which may be related to the dual inhibition of TGF-β1/Smad3/CTGF signaling pathway by upregulating atrial natriuretic peptide and BNP levels and blocking RAAS activation.

Objective:To investigate the effects and mechanisms of cyclic adenosine monophosphate (cAMP) on cisplatin-induced acute kidney injury (AKI).Methods:GSE227970 dataset derived from human renal tubular epithelial cells (HK-2 cells) in the GEO database was downloaded, and Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis were performed in normal and cisplatin-damaged cells. Eighteen 8-week-old male C57BL/6J mice with body weight of (22±2) g were randomly divided into normal group, cisplatin group and cAMP group according to random number table method, with 6 mice in each group. cAMP group was intraperitoneally injected with 30 mg/kg glumine cyclic adenosine monophosphate, while normal group and cisplatin group were intraperitoneally injected with the same volume of 0.9% sodium chloride solution for 10 consecutive days. The cisplatin and cAMP groups were intraperitoneally injected with 20 mg/kg cisplatin once on the 8th day. The body weight and kidney weight of mice were weighed, and kidney weight to body weight ratio was calculated. The blood urea nitrogen (BUN) and serum creatinine (Scr) in mice were detected. HE staining was used to evaluate the degree of renal injury. Western blotting was used to detect the protein expression of AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) signaling pathway in renal tissues. In vitro experiments, HK-2 cells were set up in normal group, cisplatin group, cAMP group and cAMP+AMPK inhibitor group. Immunofluorescence was used to detect the protein expression of phosphorylated (p)-AMPK in HK-2 cells. Adenosine triphosphate content and ratio of NAD + to NADH in cells were detected. Flow cytometry was used to detect cell apoptosis. Results:Biological signal analysis showed that axon guidance, Ras-related protein 1 signaling pathway and cAMP signaling pathway were significantly changed in cisplatin group. The body weight, kidney weight and kidney weight/body weight ratio in cisplatin group were significantly lower than those in normal group (all P<0.05). However, after cAMP treatment, kidney weight was significantly higher compared with cisplatin group ( P<0.05), and body weight and kidney weight/body weight ratio also increased, but the differences were not statistically significant (both P>0.05). BUN, Scr and renal tubular injury score in cisplatin group were significantly higher than those in normal group (all P<0.05). After cAMP treatment, BUN, Scr and renal tubular injury score were significantly lower than those in cisplatin group (all P<0.05). Western blotting results showed that cAMP treatment could significantly increase the decreased AMPK/ACC signaling pathway protein in the renal tissues of cisplatin-induced mice (all P<0.05). In vitro experiments, immunofluorescence detection showed that the expression of p-AMPK protein in cisplatin-induced HK-2 cells decreased, and the addition of cAMP increased the expression of p-AMPK protein in cisplatin-induced HK-2 cells (all P<0.05). cAMP treatment could alleviate cisplatin-induced injury in HK-2 cells, restore the reduction of adenosine triphosphate content and NAD +/NADH ratio, and reduce the apoptosis induced by cisplatin (all P<0.05), while AMPK inhibitor could eliminate the protective effect of cAMP on cisplatin-induced injury in HK-2 cells. Conclusion:cAMP can play a protective role in renal injury caused by cisplatin, and its mechanism may be related to activation of AMPK/ACC signaling pathway.

Objective:To investigate the effects and mechanisms of cyclic adenosine monophosphate (cAMP) on cisplatin-induced acute kidney injury (AKI).Methods:GSE227970 dataset derived from human renal tubular epithelial cells (HK-2 cells) in the GEO database was downloaded, and Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis were performed in normal and cisplatin-damaged cells. Eighteen 8-week-old male C57BL/6J mice with body weight of (22±2) g were randomly divided into normal group, cisplatin group and cAMP group according to random number table method, with 6 mice in each group. cAMP group was intraperitoneally injected with 30 mg/kg glumine cyclic adenosine monophosphate, while normal group and cisplatin group were intraperitoneally injected with the same volume of 0.9% sodium chloride solution for 10 consecutive days. The cisplatin and cAMP groups were intraperitoneally injected with 20 mg/kg cisplatin once on the 8th day. The body weight and kidney weight of mice were weighed, and kidney weight to body weight ratio was calculated. The blood urea nitrogen (BUN) and serum creatinine (Scr) in mice were detected. HE staining was used to evaluate the degree of renal injury. Western blotting was used to detect the protein expression of AMP-activated protein kinase (AMPK)/acetyl-CoA carboxylase (ACC) signaling pathway in renal tissues. In vitro experiments, HK-2 cells were set up in normal group, cisplatin group, cAMP group and cAMP+AMPK inhibitor group. Immunofluorescence was used to detect the protein expression of phosphorylated (p)-AMPK in HK-2 cells. Adenosine triphosphate content and ratio of NAD + to NADH in cells were detected. Flow cytometry was used to detect cell apoptosis. Results:Biological signal analysis showed that axon guidance, Ras-related protein 1 signaling pathway and cAMP signaling pathway were significantly changed in cisplatin group. The body weight, kidney weight and kidney weight/body weight ratio in cisplatin group were significantly lower than those in normal group (all P<0.05). However, after cAMP treatment, kidney weight was significantly higher compared with cisplatin group ( P<0.05), and body weight and kidney weight/body weight ratio also increased, but the differences were not statistically significant (both P>0.05). BUN, Scr and renal tubular injury score in cisplatin group were significantly higher than those in normal group (all P<0.05). After cAMP treatment, BUN, Scr and renal tubular injury score were significantly lower than those in cisplatin group (all P<0.05). Western blotting results showed that cAMP treatment could significantly increase the decreased AMPK/ACC signaling pathway protein in the renal tissues of cisplatin-induced mice (all P<0.05). In vitro experiments, immunofluorescence detection showed that the expression of p-AMPK protein in cisplatin-induced HK-2 cells decreased, and the addition of cAMP increased the expression of p-AMPK protein in cisplatin-induced HK-2 cells (all P<0.05). cAMP treatment could alleviate cisplatin-induced injury in HK-2 cells, restore the reduction of adenosine triphosphate content and NAD +/NADH ratio, and reduce the apoptosis induced by cisplatin (all P<0.05), while AMPK inhibitor could eliminate the protective effect of cAMP on cisplatin-induced injury in HK-2 cells. Conclusion:cAMP can play a protective role in renal injury caused by cisplatin, and its mechanism may be related to activation of AMPK/ACC signaling pathway.

Objective To investigate the intervention effect of sacubitril/valsartan(Sac/Val)on doxorubicin-induced heart failure(HF)in rabbits and its regulative effect on the transforming growth factor β1(TGF-β1)/SMAD family member 3(Smad3)/connective tissue growth factor(CTGF)signaling pathway.Methods Thirty-eight male New Zealand rabbits were subjected,and 8 of them were randomly assigned into a control group.The other 30 rabbits were injected with doxorubicin to establish a rabbit HF model,and finally,there were 6 rabbits in a model group,7 in a valsartan group,and 7 in a Sac/Val group.After 8 weeks of intervention,echocardiography[left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD)],and myocardial histopathologic observation(HE staining,Masson staining)were performed,and collogen volume fraction(CVF)was calculated.The levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),soluble growth stimulation expressed gene 2(sST2),galectin-3(Gal-3)were detected.Activities of renin-angiotensin-aldosterone system(RAAS)and levels of atrial natriuretic peptide,B-type brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP)and protein kinase G(PKG)were measured.The expression of α-smooth muscle actin(α-SMA),typeⅠ collagen,TGF-β1,Smad3,recombinant SMAD family member 7(Smad7)and CTGF in the myocardial tissues were detected.Results Compared with the control group,the model group exhibited significantly lower LVEF and LVFS and decreased expression of Smad7,higher LVEDD and LVESD(P＜0.01).The CVF of each group was(7.15±0.82)％、(43.20±5.09)％、(29.53±4.05)％、(22.48±2.93)％.Valsartan and Sac/Val treatment resulted in obvious increases in LVEF and LVFS,up-regulation of Smad7,decreased in LVEDD,LVESD and CVF,reduced levels of NT-proBNP,sST2,Gal-3,AngⅡ,aldosterone,atrial natriuretic peptide,BNP,cGMP and PKG,and down-regulation of α-SMA,collagen I,TGF-β1,Smad3 and CTGF when compared with the model group(P＜0.01).Sac/Val treatment showed better effects in above indicators than simple valsartan treatment(P＜0.01).Conclusion Sac/Val can reduce myocardial fibrosis and improve cardiac function in doxorubicin-induced HF rabbits,which may be related to the dual inhibition of TGF-β1/Smad3/CTGF signaling pathway by upregulating atrial natriuretic peptide and BNP levels and blocking RAAS activation.

Objective To analyze the effects of GATA binding protein 3(GATA3)mediated mini RNA-21(miR-21)/phosphatase and tensin homologue(PTEN)axis missing from human chromosome Chromosome 10 on the proliferation and invasion of endometrial cancer cells.Methods HEC-1-A cells were transfected and divided into control group,GATA3 empty plasmid group,GATA3 overexpression plasmid group,GATA3 siRNA negative control group,and GATA3 siRNA group.Detect the expression levels of GATA3,miR-21,PTEN,proliferation,apoptosis rate,migration,and invasion in each group of cells.Results Compared with the hEEC group,the expression levels of GATA3 and miR-21 in cells of the HEC-1-A group,HEC-1-B group,and Ishikawa group increased,while the expression levels of PTEN decreased(P<0.05).Compared with the GATA3 empty plasmid group,the GATA3 overexpression plasmid group showed an increase in GATA3,miR-21 mRNA expression,pro-liferation rate,migration distance,number of invading cells,and Vimentin levels,while the PTEN mRNA expression,apoptosis rate,Caspase-9,Bax,and E-cadherin levels decreased(P<0.05);Compared with the GATA3 siRNA negative control group,the GATA3,miR-21 mRNA expression,proliferation rate,migration distance,number of invading cells,and Vimentin level decreased,while the PTEN mRNA expression,apoptosis rate,Caspase-9,Bax,and E-cadherin levels increased(P<0.05).Conclusion Downregulation of GATA3 expression can regulate the miR-21/PTEN axis,slow down the proliferation of HEC-1-A cells,and promote apoptosis of HEC-1-A cells.

Objective To study the coordination and function of a laparoscopic assistant in laparoscopic pancreaticoduodenectomy (LPD).Methods A retrospective analysis was conducted on 101 patients who underwent LPD at the Department of Hepatobiliary Surgery,Hunan Provincial People's Hospital,from January 2014 to March 2017.The study aimed to study the coordination and function of a laparoscopic assistant.Results LPD was successfully completed in all the 101 patients.There was no conversion to open surgery.The operation time was (326.0 ± 55.6) min,and the resection time was (174.4 ± 42.5) min.The digestive tract reconstruction time was (101.0 ± 21.4) min.The time of pancreaticojejunostomy was (40.5 ± 8.7) min.The time of gastrointestinal anastomosis was:(26.3 ± 5.5) min.The time of biliary anastomosis was (24.4 ± 6.5) min.The intraoperative bleeding was (175.6 ± 41.1) ml.Postoperative pathological data showed that 27 patients (26.7％) had distal common bile duct cancer,23 patients (22.8％)ampullary carcinoma,39 patients (38.6％) duodenal papillary carcinoma,and 12 patients (11.9％) pancreatic ductal adenocarcinoma.The tumor diameter was (2.3 ± 1.3) cm,and the number of resected lymph nodes was (16.7 ±4.2).The number of positive lymph nodes was 1.3 ± 1.1.The length of postoperative hospital stay was 14.8 (8 ～ 29) d.Twenty-three patients developed postoperative pancreatic fistula,including 17 patients (16.8％) with a biochemical fistula,5 patients (5.0％) with a grade B pancreatic fistula,and 1 patient (1.0％) with a grade C pancreatic fistula.There were 2 patients (3.0％) with bile leakage,7 patients (6.9％) with intra-abdominal bleeding,4 patients (4.0％) with delayed gastric emptying,6 patients (5.9％) with abdominal infection,3 patients (3.0％) with pulmonary infection,2 patients (2.0％)with intestinal obstruction,3 patients (3.0％) required a repeated operation,and 1 patient (1.0％) with death in hospital within 30 days after surgery.Conclusions The laparoscopic assistant should have the perspective of "one axis,two sides and four regions" in LPD,and warn the operator to ensure the safety and fluency of the operation by clearly exposing important blood vessels and organs when performing the Kocher incision and when dissecting the key parts such as the dangerous triangle of the uncinate process.During anastomosis,the laparoscopic assistant should appropriately adjust the distance of vision,clearly reveal the surgical field of the anastomotic area,and help the surgeon in improving the precision of the suture and the quality of the anastomosis.

Objective: To observe the protective effect of salidroside pretreatment on rabbit heart after limb ischemia/reperfusion (I/R). Methods: A total of 24 New Zealand rabbits were randomly and equally divided into sham operation group, I/R + placebo group （I/R group）and salidroside pretreatment group（salidroside group）. Before establishment of rabbit models of limb I/R, salidroside group received salidroside injection via ear marginal vein, and I/R group received saline injection, once a day for three days. After model establishment, echocardiography was used to evaluate rabbit cardiac function of each group 4h after reperfusion, including left ventricular end-systolic dimension (LVESd), left ventricular end-diastolic dimension (LVEDd), left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS). Blood sample was taken to measure serum cardiac troponin I (cTnI) and tumor necrosis factor (TNF)-α. Some ventricular tissue homogenates were taken to measure levels of superoxide dismutase (SOD) and malonyl diadehyde (MDA). Then heart sample was taken to receive pathological examination. Results: Compared with I/R group 4h after reperfusion, there were significant decrease in LVESd [(0.69±0.07) mm vs. (0.62±0.05) mm] and significant increase in LVEF [(64.6±3.4) % vs. (72.1±3.6) %], FS [(34.2±3.2) % vs. (41.7±3.4) %] (P<0.05 all), but these indexes of salidroside group were all no significant different than those of sham operation group (P>0.05). Compared with I/R group, there were significant decrease in cTnI [(5.24±0.34) μg/ml vs. (1.06±0.12) μg/ml], MDA [(8.92±2.18) μmol/L vs. (6.79±1.43) μmol/L] and TNF-α [(37.43±10.02) pg/ L vs. (19.73±6.31) pg/ L], and significant increase in SOD level [(16.61±3.75) U/ml vs. (22.26±4.73) U/ml] in salidroside group (P<0.05 all). Pathological results indicated that injury degree in salidroside group was significantly attenuated than that of I/R group. Conclusion: Salidroside pretreatment could protect cardiac function and relieve rabbit cardiac injury after limb ischemia/reperfusion.

Objective To provide anatomical basis for lateral antebrachial neurocutaneous flap pedi-cled with inferior cubital artery perforator in repairing tissue defects around elbow joint. Methods Thirty embalmed upper limbs of adult cadavers perfused with red latex were used for this study, and followings were observed:①The course and distribution of lateral antebrachial cutaneous nerve; ②Anastomoses between inferior cubital artery and nutrient vessels of lateral antebrachial cutaneous nerve. Mimic operation was performed on other side of fresh specimen. Results ①The main trunk of lateral antebrachial cutaneous nerve (LACN) lined in the radial forearm and distributed in the 1/3 region of lateral forearm. ①The nutritional vessels of the flap were plurisegmental and polyphyletic. The inferior cubital artery which was relatively constant reached to skin through "V"-shaped peak formed by communicating branches of cephalic vein and deep venous system. They also gave off large number of small veins, which closely aligned with perineural branches and neural stem vascular chain of lateral antebrachial cutaneous nerve. Conclusion The lateral antebrachial neurocutaneos flap pedicled with inferior cubital artery perforator can be formed to repaire tissue defects around elbow joint.

Objective To observe the anatomy of the perforator flap of the posterolateral midforearm. Methods Lateral condyle of the humems wag taken as the observation mark on 30 specimens of adult upper limb perfused with red latex.The surgical magnifier Wag used to obse~e the origin,branches and distribution of the perforating branches of the posterolateral midforearm as well as alanagtomosis between perforating branches and peripheral vessels.Mimic operation WaS performed on the two sides of the fresh specimen.Results The perforating branches of the posterolateral midforearm originated from the radial musculoculancous branches of the posterior interosseous artery,the intermuscular branches of the radial artery and the direct periosteal branch of the radial artery had relatively stable location of piercing the deep fascia.Then,the perforating branches of the posterolateral midforearm pagsed through the deep fascia to the subcutaneous part among the spatium intermusculare of extensor digitorum and extensor carpi radialis brevis,supinator and abductor pollicis longus(within 12.5-15.8 cm below the lateral condyle of the humerus).Large number of small blood Vessels were also separated and closely aligned with the musculoculancous branches vascular,perineural and neural stem vascular chain of lateral branches of posterior antebrachial cutaneous nerve.Then,the vascular plexus was formed along the spatium intermusculare and lateral branches of posterior antebrachial cutaneous nerve longitudinal axis between extensor digitorum and extensor carpi radialis brevis. Conclusion The axial pattern flaps or cross-regional blood supply skin flap pedicled with the perforating branches of the posterolateral midforearm Can be formed to repair the soft tissue defect of tlle forearm and wrist.