Hepatocyte transplantation is an effective method for the end stage liver diseases. This procedure includes hepatoeyte isolation, culture, cryopreservation and so on. The cryopreservation is an important aspect for hepatocyte transplantation, which determines the viability after the resuscitation. The hepatocyte cryopreservation will be summarized in this passage.

Due to the increasing use of hepatoeytes,the demand of hepatoeytes is increasing consistently in basic clinical research.The application of rational and efficient method of hepatoeyte isolation is a prereq-uisite for its application.At present,there are many methods of hepatoeyte isolation.This review summarizes the latest progress on them.

Objective The aim of this study was to exam by image for APBJ, and to further establish the principles in the treatment of APBJ. Methods From December 1979 to October 2001, 64 adult patients with APBJ underwent surgical treatment in our hospital . Cholangiopancreatographic results and surgical treatment outcomes of these patients were reviewed and analyzed. Results Of the total 64 patients 28, 32 and 4 were typed as P - C(43. 8% ) ,C - P(50% ) and Common Channel(6. 2% ) distinctively. Conclusions Patients with APBJ are often associated with biliary tract and pancreatic diseases, early discovery and appropriate surgical treatment could avoid most of the serious complications.

Objective The value of endoscopic retrograde choledocho－ pancreatography( ERCP ) to detect narrow－ winding cystic duct and the effect of surgery were studied .Methods The clinical presentations, pathogenesis, diagnostic procedures and outcome of surgery were collected and analysed in 17 patients with this change. All had preoperative ERCP as well as intraoperative cholangiography. The operation was simple cholecystectomy. Results Satisfaction was obtained througthout the followup period for 2～ 5 years . Conclusion Narrow－ winding cystic duct is not a rare biliary disorder with high rate of confirmation by ERCP and fine result following cholecystectomy.

Objective:To investigate the protective effect of schisandrin B (SchB)on the cerebral ischemia reperfusion injury of the rats and the influence in HSPA12B/PI3K/Akt signaling pathway,and to explore the mechanisms.Methods:130 SD rats were divided into sham group,cerebral ischemia reperfusion injury model group (model group),low dose of SchB group (SchB 3 mg· kg-1 ,SchB1 group),middle dose of SchB group (SchB 10 mg·kg-1 ,SchB2 group)and high dose of SchB group (SchB 30 mg·kg-1 ,SchB3 group)(n=26).The rats in sham group didn’t plug lines;the rats in model were used to establish ischemia reperfusion models;the rats in SchB1 ,SchB2 and SchB3 groups were firstly pretreated with different doses of SchB for 7 d,and then they were used to build cerebral ischemia reperfusion injury models.The nerve dysfunction of rats was evaluated by neurologic deficit score.The cerebral edema was detected by measuring the content of water in brain tissue.The morphological changes of brain tissue were observed by toluidine blue staining.The levels of nuclear factor kappa B (NF-κB), tumor necrosis factor-α(TNF-α),interleukin-1 (IL-1)and interleukin-6 (IL-6)in the brain tissue were detected by ELISA.Western blotting method was used to detect the protein expression levels of heat shock protein A12B (HSPA12B ), serine-threonine kinase (Akt ) and phosphorylation serine-threonine kinase (p-AKT ). Results:Compared with sham group,the neurologic deficit score of rats in model group was significantly increased (P <0.01),and the content of water in brain tissue was increased (P < 0.01 );the brain tissue structure was loosened,and the mesenchyme appeared edema;the NF-κB,TNF-α,IL-1,and IL-6 levels were increased (P <0.01),and the expression levels of HSPA12B and p-Akt proteins were decreased (P <0.01).Compared with model group,the neurologic deficit scores of the rats in SchB1 ,SchB2 ,and SchB3 groups were decreased (P <0.01),and the contents of water in brain tissue of the rats in SchB2 and SchB3 groups were decreased (P <0.05);the edema of nerve cells was alleviated,and the cavities were reduced;the NF-κB,TNF-α,IL-1,and IL-6 levels were decreased (P <0.05 or P <0.01),the expression levels of the HSPA12B protein in SchB2 ,and SchB3 groups were increased (P <0.05),and the p-Akt protein expression levels of the rats in SchB1 ,SchB2 ,and SchB3 groups were increased (P <0.01).Conclusion:SchB could protect the cerebral ischemia reperfusion injury of rats,its mechanism may be related to regulating the HSPA12B/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.

Objective To detect the expression of p53 and k-ras in benign and malignant gallbladder diseases.Method ABC immunohistochemistry was used to detect p53 and k-ras protein in 101 specimens of chronic cholecystisis with cholecystolithiasis(group A),20 with cholesterol polyp(group B),22 with adenoma of gallbladder(group C),25 with primary gallbladder cancer(PGC)(in which there were 5 with atypical hyperplasia and carcinoma in situ)(group D)in this study.The relationship between p53 or/and k-ras expression and clinicopathologic data were analyzed.Resuit p53 and k-ras expressions were detected in both benign and malignant specimens,with the positive rates increasing from cholesterol polyp,to chronic cholecystisis to adenoma of gallbladder to carcinoma in situ and PGC(p53 was 62.8% in patients with benign lesions,and 92.0%in malignant lesion x2=8.16,P=0.004;k-ras was positive in 66.4% patients with benign lesions,and 92.0% in malignant lesions x2=6.65,P=0.01).In benign lesions the p53 expressions in group C was higher than that in group A and B(x2=6.90 and 6.02,P=0.009 and 0.014).There was higher expression of p53 in chronic chylecystisis with a history more than 10 years than those less than 10 years(80.0%vs.52.9%,x2=8.28,P=0.004).k-ras in cases less than 60 years old was higher than those after 60 years of age(70.4%vs.46.8%,x2=5.79,P=0.016).Conclusion The higher expression rate of p53 in cholecyst adenoma may predict carcinogenesis,while k-ras is negatively correlated with patients'age,which may bespeak the development of carcinoma in patients younger than 60 years old.

Objective:To investigate the effect of jejunal loop fixation in the reoperation for anastomotic stricture after cholangiojejunostomy.Methods:From Jan 2016 to Jan 2020, clinical data of 15 patients undergoing reoperations for anastomotic stenosis was analyzed retrospectively.Original anastomosis was removed and Roux-en-Y cholangiojejunostomy and jejunal loop fixation were performed in all cases. Two different types of jejunal loop fixation were used. One with a blind loop of 10-15 cm at the proximal side of the anastomosis, which is fixed with a mark of the abdominal wall under the xiphoid process. The second is to mark and fix the jejunal side wall and the right abdominal wall about 10 cm from the distal side of the anastomosis. A T tube is placed in the intestine through the central part of the fixed intestinal wall.Results:Five cases underwent blind loop fixation, 10 cases underwent lateral wall fixation. There were 3 complications after operation, including 1 case of bile fistula, 1 case of incision infection, 1 case of abdominal hemorrhage, all were cured by conservative treatment, and there was no perioperative death. One case had postoperative anastomotic stenosis in the follow-up. The patient underwent jejunal puncture under local anesthesia, and was cured by percutaneous choledochoscopy.Conclusions:The possibility of restenosis should be considered in the reoperation of anastomotic stenosis. The jejunal loop should be fixed and marked during the operation in high risk patients. Once the anastomotic stricture recurred, choledochoscopy could be performed by puncture and dilation of fixed loop of intestine to avoid reopen surgery.

OBJECTIVETo explore the method for improving the aging resistance of seeds and seedlings of Perilla frutescens through study on seed germination and physiological characteristics of P. frutescens seedlings.

METHODSeveral physiological indexes of P. frutescens seeds treated by different concentrations of ZnSO4 and PEG were measured. And other indexes like the activities of malondialdehyde (MDA), superoxide (SOD), peroxidase (POD) and catalase (CAT) were also determined.

RESULTThe germination indexes of P. frutescens aging seeds treated by different concentrations of ZnSO4 and PEG were all increased. And the seeds that treated by ZnSO4 (600 mg x L(-1)) and PEG (20%) showed the most significantly increase in every index. The germination vigor were 64.7% and 66.8%, the germination rate were 78.7% and 79.4%, the germination index were 11.8 and 12.2, the vigor index were 0.091 1 and 0.0939 respectively. The content of MDA was decreased under different treatment. The activities of three enzymes include SOD, POD and CAT were increased by different treatment of ZnSO4 (0.28, 4.71, 3.82 U x mg(-1) respectively) and PEG (0.29, 4.93, 4.18 U x mg(-1) respectively).

CONCLUSIONZnSO4 with concentration of 600 mg x L(-1) and PEG with concentration of 20% could significantly alleviate the damages to the seeds and seedlings of P. frutescens by aging and promote the aging resistance of the seeds and seedlings.

Catalase ; metabolism ; Germination ; drug effects ; Malondialdehyde ; metabolism ; Perilla frutescens ; drug effects ; enzymology ; physiology ; Peroxidase ; metabolism ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Polyethylene Glycols ; pharmacology ; Seedlings ; enzymology ; Seeds ; enzymology ; physiology ; Zinc Sulfate ; pharmacology

OBJECTIVETo find a method for improving the salt resistance of seeds and seedlings for Perilla Frutescens under NaCl stress, seed germination and physiological characteristics of P. frutescens seedlings were studied.

METHODSeveral physiological indexes of P. frutescens seeds treated with different concentrations of Ca2+, 5-aminolevulinic acid (ALA), salicylic acid (SA) and spermidine (Spd) under NaCl stress like the germination vigor, germination rate, germination index and vigor index were measured. And other indexes like the biomass of the seedlings, the content of malondialdehyde (MDA) in leaves, the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT) were also measured.

RESULTThe germination of P. frutescens seeds under NaCl stress (100 mmol x L(-1)) was inhibited obviously. But after the treatment with Ca2+, ALA , SA and Spd, all germination indexes were increased. Ca2+ (10 mmol x L(-1)), ALA (100 mg x L(-1)), SA (50 mg x L(-1)) and Spd (0.25 mmol x L(-1)) could obviously alleviate the damage of salt stress to the seeds of P. frutescens. ALA (100 mg x L(-1)) significantly increased all indexes. The germination vigor was 65.3%, the germination rate was 89.7%, the germination index and vigor index were 15.2 and 0.1238, respectively. All treatments decreased the content of MDA in leaves. The activities of three enzymes including SOD, POD and CAT were all increased. ALA (100 mg x L(-1)) had the enzymes activity reach the maximum with 0.72, 6, 82 and 5.64 U x mg(-1), respectively.

CONCLUSIONCa2+ ALA , SA and Spd with appropriate concentration could significantly alleviate the damages to the seeds and seedlings of P. frutescens under NaCl stress and promote the salt resistance of the seeds and seedlings.

Aminolevulinic Acid ; pharmacology ; Calcium ; pharmacology ; Catalase ; metabolism ; Dose-Response Relationship, Drug ; Germination ; drug effects ; Malondialdehyde ; metabolism ; Perilla frutescens ; drug effects ; enzymology ; metabolism ; physiology ; Peroxidase ; metabolism ; Salicylic Acid ; pharmacology ; Seedlings ; drug effects ; enzymology ; metabolism ; physiology ; Sodium Chloride ; pharmacology ; Spermidine ; pharmacology ; Stress, Physiological ; drug effects ; Superoxide Dismutase ; metabolism

OBJECTIVEIn order to find a method for improving the salt resistance of seeds and seedlings for Perilla frutescens under NaCl stress, seed germination and physiological characteristics of P. frutescens seedlings were studied.

METHODSeveral physiological indexes of P. frutescens seeds treated by Ca2+ and sodium nitroprusside (SNP) under NaCl stress like the germination vigor, germination rate, germination index and vigor index were measured. And other indexes like the biomass of the seedlings, the content of malondialdehyde (MDA) in leaves, the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT) were also measured.

RESULTThe germination of P. frutescens seeds under NaCl stress was inhibited obviously. But after the treatment with Ca2+ and SNP, all of the germination indexes increased. And the seeds that treated with SNP + Ca2+ had the most significantly increase in all indexes. The germination vigor was 65.1%, the germination rate was 89.3%, the germination index and vigor index were 13.9 and 0.1109, respectively. The content of MDA decreased after the treatment. The activities of three enzymes include SOD, POD and CAT were increased by the treatment and get the maximin 0.84, 5.71 and 4.92 U x mg(-1) respectively. And the EGTA showed an obvious inhibition to the effect of SNP on P. frutescens.

CONCLUSIONSNP (0.1 mmol x L(-1)) and Ca2+ (10 mmol x L(-1)) could significantly alleviate the damages to the seeds and seedlings of P. frutescens under NaCl stress, and promote the salt resistance of the seeds and seedlings. These results might suggest that exogenous NO might enhance P. frutescens salt resistance and alleviate salt injury possible by enhancing Ca2+ influx by activating Ca2+ channels and improving concentration of Ca2+ of P. frutescens seedlings.

Calcium ; pharmacology ; Catalase ; metabolism ; Germination ; drug effects ; Nitroprusside ; pharmacokinetics ; Perilla ; drug effects ; enzymology ; physiology ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seedlings ; drug effects ; enzymology ; metabolism ; physiology ; Sodium Chloride ; metabolism ; Stress, Physiological