Objective The intervention effect of Huoxue Jiedu Huayu Recipe on the pyroptosis of vascular smooth muscle cells(VSMC)on the contralateral renal vessel of 6-month unilateral ureteral obstruction(UUO)rat model was observed,and part of the treatment mechanism of Huoxue Jiedu Huayu Recipe on chronic kidney disease(CKD)was investigated.Methods Forty male wistar rats were randomly divided into 4 groups:the sham operation(Sham)group,the unilateral ureteral obstruction(UUO)group,the eplerenone treatment(EPL)group and the Huoxue Jiedu Huayu Recipe intervention(HJHR)group,10 rats in each group.The EPL group was given eplerenone and the HJHR group was given traditional Chinese medicine.The right kidney was collected after 6 months.Pathological changes were assessed by HE staining and Masson staining.The expressions of Cysteinyl aspartate specific proteinase1(Caspase-1),Interleukin-1β(IL-1β),Serum and glucocorticoid-induced kinase 1(SGK1)and nuclear factor κappa-B(NF-κB)were detected by immunohistochemistry.The DNA damage in renal VSMC was examined by TUNEL staining.Rat VSMCs were used for in vitro experiments.The cells were divided into 4 groups using the random number method:the control(CON)group,the aldosterone(ALD)group,the aldosterone plus eplerenone treatment(EPL)group and the aldosterone plus Huoxue Jiedu Huayu Recipe treatment(HJHR)group.The ALD group was given aldosterone stimulation for 24 h.The EPL group and the HJHR group were given eplerenone and rat serum containing 10%Huoxue Jiedu Huayu Recipe pretreatment before aldosterone stimulation,respectively.The apoptosis rate was detected by flow cytometry.Perforation of gasdermin D(GSDMD)and nuclear translocation of NR3C2(Encode MR)in cell membranes were examined by immunofluorescence.The protein expression of inflammation and pyroptosis-related signaling molecules were examined by western blotting.The mRNA expression of pyroptosis signaling pathway were detected by qRT-PCR.Results Compared with the Sham group,DNA damage of renal blood vessels and VSMCs was increased in UUO group,and expressions of Caspase-1,IL-1β,SGK1 and NF-κB were increased.Compared with the UUO group,the DNA damage of renal blood vessels and VSMCs in EPL and HJHR groups was reduced,and the expressions of Caspase-1,IL-1β,SGK1 and NF-κB were decreased.Compared with the CON group,the ALD group showed pyroptosis and DNA damage of rat VSMC.The expression of NR3C2,NLRP3,Caspase-1,GSDMD,IL-1β,SGK1 and NF-κB was up-regulated.Compared with the ALD group,the pyroptosis and DNA damage were alleviated,and the expression of NR3C2,NLRP3,Caspase-1,GSDMD,IL-1β,SGK1 and NF-κB was decreased in EPL and HJHR groups.Conclusion Huoxue Jiedu Huayu Recipe reduce the vascular damage of the contralateral kidneys of UUO by inhibiting the activation of the MR/NLRP3 inflammasome pathway and inhibiting the pyroptosis of VSMC.

The dual luciferase reporter gene system provides sensitive readout, while it relies on a constitutively-expressed control gene for readout normalization. However, most standard control reporter genes are not constitutively expressed under all conditions. Here, we report an effective method to construct a control reporter plasmid for the dual luciferase reporter gene system that would be suitable for hormone research in silkworm cell lines. First, we modified BmVgP78M, a stably-expressed constitutive promoter in silkworm cells by mutating its hormone-related element. Then, we constructed the pRL-VgP78M control reporter plasmid by replacing the SV40 promoter and chimeric intron sequences in pRL-SV40 with the BmVgP78M sequence. Finally, we confirmed that the pRL-VgP78M control reporter plasmid could be stably expressed in silkworm cell lines via cell transfection experiments, and it was unresponsive to the induction of ecdysone, juvenile hormone, or their transcription factors. We thus obtained a control reporter plasmid pRL-VgP78M that could be expressed stably and moderately in silkworm cells. It can be readily used as the control reporter plasmid of the dual luciferase reporter gene system for hormone research in silkworm cell lines. It will also provide a reference for construction of control reporter plasmids of dual luciferase reporter gene systems that are adaptable to cell lines isolated from other species.

Aim To observe the effect of eplerenone(EPL) and Chinese decoction on cell apoptosis in obstructive nephropathy rats.Methods Sixty male Wistar rats were randomly divided into sham group, UUO group, EPL group and ZY group(n=15).Except sham group, the rats in the other groups were ligated with unilateral ureteral obstruction(UUO) for renal interstitial fibrosis model.The rats were treated with eplerenone at 100 mg·kg-1·d-1 added to diet in EPL group, and orally 13.7 g·kg-1·d-1 decoction of Chinese medicine in ZY group.The kidneys were harvested on 14th day, the number of renal cell apoptosis were detected by TUNEL, and serum aldosterone and 8-OhdG were detected with radioimmunoassay and ELISA.Caspase-12, caspase-9, Bax and Bcl-2 were examined by immunohistochemistry and Western blot.Results The levels of serum aldosterone, serum and urine 8-OhdG and the number of positive apoptotic cells increased significantly in UUO rats compared with Sham group.The overexpression of caspase-9, caspase-12 and Bax and down-regulated Bcl-2 were obvious in UUO group(P<0.01).The level of 8-OhdG, expression of caspase-9, caspase-12 and Bax were down-regulated, and Bcl-2 expression was up-regulated in eplerenone and Chinese decoction treated rats(P<0.01).Conclusion Eplerenone and Chinese decoction could inhibit cell apoptosis induced by oxidative damage after UUO via caspases and(or) Bax pathway.

Objective To evaluate the effect of dexmedetomidine on renal functions during the anesthesia of liver transplantation patients.Methods Forty patients (male 31 cases,female 9 cases, aged 40-60 years,ASA grade Ⅱ or Ⅲ)received liver transplantation were randomly divided into two groups(n =20):dexmedetomidine group (group D)and normal saline group (group C).Patients in the group D received a loading dose of dexmedetomidine (0.5 μg/kg within 10 min)and a continuous infusion of dexmedetomidine (0.4 μg·kg-1 ·h-1 )until the end of surgery,while patients in group C received saline.Central venous blood and urine were collected after induction of anesthesia (T1 ),the anhepatic phase of liver 30 min (T2 ),new liver stage 30 min (T3 ),new liver stage 6 h (T4 ),postop-erative 24 h (T5 )and postoperative 1 week (T6 )to detect the serum cystatin C,endogenous creati-nine clearance rate,blood urea nitrogen,blood creatinine,urinary NAG enzyme,urinary albumin, and red blood cells.The use of vasopressors and diuretics,blood loss,fluid,urine,and blood transfu-sion (including RBC,fresh frozen plasma,and platelets)were all recorded.Results Compared with T1 ,serum cystatin C,blood urea nitrogen,serum creatinine of group D increased significantly and en-dogenous creatinine clearance rate reduced significantly at T3 ,T4 (P < 0.05 ).Microalbuminuria in-creased at T3-T5 (P <0.05).Serum cystatin C,blood urea nitrogen,serum creatinine of group C in-creased significantly and endogenous creatinine clearance rate reduced significantly(P < 0.05 ).Com-pared with group C,serum cystatin C,blood urea nitrogen,serum creatinine of group D reduced signif-icantly at T3-T5 and endogenous creatinine clearance rate increased(P <0.05).Microalbuminuria re-duced significantly at T4 ,T5 (P <0.05 ).Perioperative use of diuretics in group D patients was less than that in group C,but the use of vasopressors in group D patients was more than that in group C (P <0.05).Urine volume in group D was more than that in group C (P < 0.05 ).There was no difference in perioperative blood loss, fluid, and blood transfusion between two groups. Conclusion Perioperative continuous infusion of dexmedetomidine might effectively alleviate acute kidney injury during operation and decrease the use of diuretics.

Aim To observe the effect of mineralocor-ticoid receptor blockade eplerenone on cell proliferation in obstructed kidney of rats. Methods Renal intersti-tial fibrotic animals were made with unilateral ureteral obstruction (UUO) and treated with eplerenone100 mg · kg - 1 · d - 1 . The kidneys were harvested on the 10th day and proliferating cell nuclear antigen ( PC-NA ), serum and glucocorticoid induced kinase-1 (SGK-1 ) and transforming growth factor-β1 ( TGF-β1 ) were detected with immunohistochemistry and Western blot. Results Renal histopathology showed large quantities extracellular matrix (ECM) accumula-tion in kidney with UUO, large numbers of inflammato-ry cells infiltrated in renal interstitium, renal tubular expansion and exfoliation of epithelial cells . The cell proliferation and ECM accumulation were inhibited in eplerenone treated rats significantly. Immunohisto-chemistry and Western blot showed that expressions of PCNA,SGK-1 and TGF-β1 were significantly up-regu-lated with UUO and down-regulated by eplerenone. Conclusion Eplerenone plays the role in inhibiting the cell proliferation and reducing ECM accumulation by down-regulating expression of SGK-1 pathway in rats with unilateral ureteral obstruction.

Objective To evaluate the blood-saving effect of prophylactic use of tranexamic acid in patients undergoing orthotopic liver transplantation.Methods Sixty ASA physical status Ⅰ-Ⅲ patients of both sexes,aged 18-60 yr,weighing 45-80 kg,scheduled for elective orthotopic liver transplantation,were randomly assigned to one of 2 groups (n =30 each) using a random number table:prophylactic use group (group P) and therapeutic use group (group T).Immediately after induction of anesthesia (T1),at 30 min of anhepatic phase (T2),and at 30 min and 2 h of neohepatic phase (T3,4),central venous blood samples were collected to determine plasma fibrinogen concentration (Fib) and platelet count,and the arterial blood samples were obtained to detect thromboelastography (TEG) parameters.In group T,when lysis after 30 min＞7.5％ and Clot Index ≤ 1.0 according to the results of TEG,which indicating that primary hyperfibrinolysis occurred,tranexamic acid 15-20 mng/kg was injected intravenously.In group P,immediately after beginning of skin incision,immediately after occlusion of portal vein,and immediately after portal vein unclamping,tranexamic acid 1 g was injected intravenously,and a single injection of tranexamic acid 15-20 mg/kg was given when primary hyperfibrinolysis occurred.The intraoperative blood loss,fluid input and output and transfusion of blood components were recorded.The duration of stay in ICU,amount of abdominal drainage during stay in ICU,volume of blood transfused within 72 h after operation,and hepatic artery and portal vein thrombosis within 1 week after operation were recorded.Results Compared with group T,the intraoperative blood loss,volume of succinylated gelatin injection transfused,and requirement for platelet and cryoprecipitate were significantly reduced,Angle at T2 and lysis after 30 min at T2,3 and maximum amplitude at T3 were increased,and no significant change was found in the duration of stay in ICU,postoperative amount of abdominal drainage and volume of blood transfused in group P.No patients developed primary hyperfibrinolysis in group P.No hepatic artery and portal vein thrombosis was detected within 1 week after operation in the two groups.Conclusion Prophylactic use of tranexamic acid can effectively prevent hyperfibrinolysis and reduce intraoperative blood loss without increasing the risk of development of thrombosis,and it provides better blood-saving effect than therapeutic use guided by TEG in patients undergoing orthotopic liver transplantation.

BACKGROUND:Osteoarthritis animal models induced by papain are commonly used for studying osteoarthritis.
OBJECTIVE:To observe the morphology of articular cartilage surface during early osteoarthritis process in rats induced by papain and L-cysteine under scanning electron microscope.
METHODS:The right knees of Sprague-Dawley rats were given 0.15 mL mixed solution of 2%papain and 0.03 mol/L L-cysteine in the ratio of 2:1. The left knees were given injections of 0.15 mL 0.9%NaCl, as the control group. Another two rats (four knees) maintained untreated, as the normal control group. Al the rats were observed with scanning electron microscope at 2, 4, 6 weeks after injection, to explore the surface morphology of medial condyle of femur.
RESULTS AND CONCLUSION:The condyles of normal and control groups revealed numerous shal ow pits on the surface of articular cartilage. At 2 weeks after injection, the surfaces of condyles were unsmooth, presenting the shrinkage and distortion. The thinner articular cartilage and local tiny cracks were seen at 4 weeks. The deep and great cracks and cartilage defect were observed on the surface of cartilage at 6 weeks. The time of 4-6 weeks is a reference point for establishing early osteoarthritis model in rats through injections of 2%papain and 0.03 mol/L L-cysteine.

Objective To discuss the diagnosis and management of metastatic adrenal carcinoma. Methods Forty-seven cases with metastatic tumor in the adrenal glands were analyzed retrospectively from December 1996 to April 2010. Lung was the most common primary tumor site (51.1 %, 24 cases), followed by the renal cell carcinoma (12.8%, 6 cases), liver cancer (10.6%,5 cases), breast cancer (8.5%, 4 cases), melanoma (4.2%, 2 cases), and other carcinoma (12.8%,6 cases). Most patients with metastatic adrenal carcinoma had no special clinic manifestation. Lesions of 36 cases were surgically removed and 11 cases gave up operation. Results The mean survival time were 33.8±4.5 months for 31 cases underwent adrenalectomy and 6.3±2.7 months for 6 cases without operation. Patients with surgically removed adrenal metastases had better survival than those without surgical resection. Conclusions Adrenal gland is a common site of metastatic carcinoma.Ultrasonographic and CT scans are important diagnosis methods for metastatic adrenal carcinoma. Operation is still effective and long-term survival may be achieved for those well selected patients, but surgical indication must be strictly monitored.

Objective To explore the incidence of tibial plateau fracture with ligament injuries and the early diagnosis and management of the condition. Methods Fifty seven cases of tibial plateau fractures which had been treated operatively from Jan. 2001 to Jun.2003 and had satisfactory reduction of the articular surface were reviewed. The stability of their knee joints was analyzed. Results The follow up lasted 6 to 30 months with an average of 15 months. The incidence of knee unstability was low. The four potential reasons for the postoperative knee stability were found to be: 1) the low prevalence of ligament injury with displaced fractures of tibial plateau; 2) incomplete ligament disruption or mild ligament injury; 3) anatomic reduction and rigid internal fixation; 4) fine and detailed plan for immobilization and functional exercise. Conclusion The prevalence of complete ligament disruption is low, and nonoperative management can result in satisfactory outcomes for tibial plateau fracture with incomplete ligament injury.