AIM: To explore the possible role of the vitamin D receptor (VDR) in the effect of 1,25-dihydroxyvitamin D 3 and its novel analogues on a human osteosarcoma cell line (HOS-8603) proliferation. METHODS: Detecting the VDR mRNA and protein expression in HOS-8603 cells by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical analysis, respectively. Using the method of transient transfection of a reporter gene (DR3-tk-CAT) for VDR to detect its function. The cell VDRas3 which stably expressed VDR antisense mRNA was used to observe the effect of 1,25(OH) 2 D 3 on the proliferation of HOS-8603 cells and the induction of p21 mRNA, one of the VDR target genes, when the VDR in the cells was blocked. RESULTS: HOS-8603 cells expressed the vitamin D receptor which functioned as a hormone-dependent transcriptional factor. When VDR in the HOS-8603 cells decreased, the cells lost the responsiveness to 1,25(OH) 2 D 3 irrespective of either the induction of p21 gene expression or the cell proliferation. CONCLUSION: The effect of 1,25(OH) 2 D 3 on the proliferation of the human osteosarcoma cell line HOS-8603 was mediated by its nuclear receptor VDR.

AIM: To investigate the effect of glucocorticoid receptor beta(GR?) on the transcriptional activation function of glucocorticoid receptor alpha(GR?) and elucidate the biological significance of glucocorticoid receptor beta. METHODS: GR? (pRSH-GR?) and GR? responsive reporter gene cat (pMMTV-cat) were cotransfected into human osteosarcoma cell line (HOS-8603). Transient transfectants were treated with dexamethasone and the activity of cat was determined by the method of ELISA. The expression p21 messenger RNA was detected by the method of reverse transcription-polymerase chain reaction. RESULTS: GR? repressed the expression of reporter gene-cat in a dose-dependent manner. The expression of messenger RNA of p21, an endogenous target gene of GR?, was also inhibited by GR?. CONCLUSION: GR? can inhibit the transcriptional activation function of GR?. GR? may be an endogenous inhibitor of GR?.

AIM: To explore the possible role of p21 wafl gene in the effect of 1,25-dihydroxyvitamin D 3 on a human osteosarcoma cell line (HOS-8603) proliferation and differentiation. METHODS: The effect of 1,25(OH) 2D 3 on the expression of p21 wafl mRNA was determined by quantitative reverse transcription-polymerase chain reaction. The human osteosarcoma cell line stably expressing the p21 wafl mRNA, which named HOS-p21 wafl , was constructed by lipofectamine transfection, and the expression of p21 wafl protein was detected by Western blot. Then the growth curve of the HOS-p21 wafl cells and the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells were investigated. RESULTS: The expression of endogenous p21 wafl mRNA in HOS-8603 cells was induced by 1,25(OH) 2D 3 in a time-dependent manner, reaching the maximum at 4 h, and remaining the inducible action for up to 24 h over basal levels. The HOS-p21 wafl cells grew much slower than the control cells, only reaching 50% of the control ones when cultured for up to 6 days. Histochemical analysis showed that the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells increased remarkably.CONCLUSION: These results suggest that the p21 wafl mRNA expression induced by 1,25(OH) 2D 3 is one of important mechanisms responsible for the cellular effects of the hormone on HOS-8603 cells.

Objective To compare the clinical effects of HC video laryngoscope and fiberoptic bronchoscope (FOB)in guidance of endotracheal intubation for patients undergoing cervical surgery.Meth-ods A total of 50 patients (ASA I or II)with cervical vertebra injury,nerve root cervical spondylopathy or cervical spondylotic myelopathy in the first affiliated hospital of Wenzhou medical university were selected, all of whom were undergone selective cervical operations between March 2014 and June 2015.The patients were randomly divided into two groups (n =25):HC video laryngoscope group (group H)and FOB group (group F).After induction of anesthesia,HC video laryngoscope and FOB were used for tracheal intuba-tion.⑴ Intubation time and success rates of intubation were recorded,and visual analogue scale (VAS) was used to evaluate the difficulty of intubation;⑵ Mean arterial pressure (MAP),heart rate (HR),and rate-pressure product (RPP)before incubation (Tb),immediate after intubation (T0),and 1min after in-cubation (T1)were recorded;(3)Intubation-related complications were recorded.Results ⑴ The dura-tion of intubation (19.7 ±7.1 )s in the group H was significantly shorter than that of group F (51.9 ± 19.2)s (P <0.05).The one-time success rate of intubation in group H (92%)was higher than that of group F (64%),with statistically significant differences (P <0.05).Intubation was easier in group H (P <0.05).⑵ MAP,HR and RPP of group H at T0 were higher than those of group F.And at T1,there were no statistical differences between groups in MAP,HR and RPP (P >0.05).⑶ The incidence of complications (sore throat)was lower in group H (P <0.05).Conclusions Compared to FOB,HC vid-eo laryngoscope-guided selective cervical operation has the characteristics of higher success rate of intubat-ion,shorter operation time,and easier operation,more stable hemodynamics and lower incidence of intuba-tion complications.Therefore,HC video laryngoscope is a safe and effective method in the intubation for pa-tients undergoing cervical surgery.

Objective To evaluate the outcomes of liver transplant recipients who received liver grafts from HBsAg positive donors in patients with hepatocellular carcinoma beyond UCSF (University of California,San Francisco) Criteria.Method The medical records of patients who underwent HBsAg-positive donor liver transplantation for hepatocellular carcinoma beyond UCSF Criteria from October 2008 to December 2012 at our hospital were analyzed retrospectively,including the existence status of HBV,graft function,tumor recurrence,and the survival after transplantation.Result A total of 20 patients were enrolled in the study.One patient lost follow-up while the remaining 19 patients had complete follow-up data.All the patients were followed up until June 2013,with a median follow-up duration of 12 months (range 2-57 months).One patient died from postoperative abdominal bleeding and multiple organ failure at post-transplantative day 21.Five patients survived up to now,including 4 cases with disease-free survival,who has been surviving for 57,35,26 and 12 months respectively.The remaining all 14 patients died from tumor recurrence at different time points after transplantation.Entecavir was used alone in 19 patients and Entecavir combined with Adefovir dipivoxilalone were used as anti-HBV therapy in the recipients.At posttransplant day 45,all the recipients were positive for serum HBsAg and negative for serum HBVDNA,and the liver enzymatic criteria,coagulation criteria and the serum bilirubin restored to normal levels or within twice the upper limit of normal levels.Throughout the follow-up period,recipients were all positive for serum HBsAg,but there was no recurrence of hepatitis B.The 1-,2-,3-and 4-year cumulative survival rate was 48.0％,35.0％,18.7％ and 18.7％ respectively.Conclusion The HBsAg positive liver may be used as a donor in liver transplantation and the graft probably works well after the operation.Liver transplantation may prolong the survival and improve the quality of life,even achieve long-term disease-free survival in patients with hepatocellular carcinoma beyond UCSF Criteria.The use of nucleotide analogue only,instead of combination with hepatitis B immune globulin,can also bring HBV well under control in liver transplant recipients with HBsAg positive donors.

Objective To compare the impact of the sirolimus and tacrolimus on the tumor recurrence after liver transplantation due to HCC beyond Milan criteria.Method Sixty-one liver transplantation recipients due to HCC beyond Milan criteria,between Jan.2008 and Apri.2012,were randomized,with the informed consent,into two different immunosuppression groups: sirolimus group (n=30) and tacrolimus group (n=31).In tacrolimus group,tacrolimus was used as the basic immunosuppressant,methylprednisolone was discontinued within one month postoperatively,and mycophenolate mofetil was used within the dosage of 1.5 g/d accordingly.In sirolimus group,the immunosuppresive scheme was the same as that of the tacrolimus group within postoperative one month,and from that,tacrolimus was transferred to sirolimus.No antineoplastic agents were given before tumor recurrence.The tumor recurrence rate and the survival rate of the recipients were compared between the two groups.Result The median follow-up duration was 35.2 months (10.3～ 60.2).The tumor recurrence rate at postoperative year 1,2,3 and 4 in the sirolimus group (13.3％,36.7％,43.3％ and 53.3％) was significantly lower than that in the tacrolimus group (38.7％,67.7％,74.2％ and 77.4％),P ＜ 0.05 for all.The one-year survival rate in the recipients postoperation had no significant difference between sirolimus group and tacrolimus group (90.0％ vs.87.1％,P=0.438).The 2-,3-and 4-year survival rate in the recipients was significantly higher in the sirolimus group (53.3％,33.3％ and 20.0％) than that in the tacrolimus group (41.9％,22.6％ and 9.7％),P ＜ 0.05 for all.The liver function and renal function of the recipients at the postoperative year 1,2,3 and 4 showed no significant difference between the two groups,P＞0.05.Conclusion In comparison with tacrolimus,sirolimus could significantly reduce the tumor recurrence rate and increase the survival rate for the liver transplant recipients due to HCC beyond Milan criteria.

Objective To investigate the effects of repetitive ultraviolet A(UVA) radiation on the endocytosis and intracellular degradation of extracellular elastin by human dermal fibroblasts(HDFs), and to explore the mechanism responsible for elastin accumulation in photoaging skin. Methods Cultured HDFs were divided into two groups:repetitive UVA radiation group treated with UVA radiation(9.9 J/cm2)once a day for seven consecutive days to establish a chronic photodamage model, and blank control group receiving no treatment. For the verification of the model, flow cytometry was performed to detect cell cycle, and senescence-associated β-galactosidase staining to determine the degree of cell senescence. Fluorescent tracer technique and conjugated polymer-based fluorescence quenching technique were conducted to observe the endocytosis and intracellular degradation of extracellular elastin in culture media by HDFs in these groups. Results Compared with the blank control group, the repetitive UVA radiation group showed increased proportions of G1-phase cells and senescent cells, which confirmed the successful establishment of chronic photodamage model in the repetitive UVA radiation group. After coculture with elastin for 24, 48 and 72 hours, the endocytosis rate of elastin was 10.0% ± 1.4%, 27.8% ± 4.2% and 39.9% ± 4.1% respectively in the blank control group, 9.9% ± 1.6%, 28.3% ± 5.1% and 42.0% ± 5.7% respectively in the repetitive UVA radiation group, with no significant difference between the two groups at the three time points (all P > 0.05). The percentage of cells showing intracellular degradation of extracellular elastin was significantly lower in the repetitive UVA radiation group than in the blank control group (4.2% ± 1.1% vs. 7.7% ± 0.9% at 24 hours, 16.5% ± 2.4% vs. 22.8% ± 1.8% at 48 hours, 26.7% ± 2.6% vs. 33.9% ± 3.1% at 72 hours, all P < 0.05). Conclusions UVA radiation at 9.9 J/cm2 for 7 consecutive days can decrease the intracellular degradation of extracellular elastin by HDFs, but has no obvious effect on endocytosis of elastin.

To summarize and explore the etiopathogenisis, clinical 'characteristics, diagnosis and treatment and prognosis of idiopathic adulthood ductopenia. We described a 27-year-old Chinese female initially manifested with jaundice, pruritus, dark urine, and pallor of the stools in October, 2002. Symptoms occurred repetitively and severed progressively, and she received therapy at the General Hospital of Chinese People's Armed Police Forces. B-ultrasound demonstrated that hepatic lesion and slightly swelling spleen. Pathological diagnosis of liver biopsy showed that she affected idiopathic adulthood ductopenia. The outcome of treatment of ursodeoxycholic acid and adrenal cortical hormone was not significant. She received orthotopic liver transplantation on June 24~(th), 2005. After liver transplantation, pruritus and jaundice were promptly disappeared. Each index of hepatic function recovered to a normal level. In March 2007, decreased dose of hormone induced an abnormal hepatic function, with mildly acute rejection. Drug dose was regulated, and hepatic function gradually returned to normal. During follow-up 51 months after the operation, she was doing well and was free of symptoms with normal liver function and no evidence of allograft dysfunction. The pathogenesis of idiopathic adulthood ductopenia is still unknown. There are still no precise and effective drugs for treatment of idiopathic adulthood ductopenia. Liver transplantation offers an effective therapy for end-stage patients with idiopathic adulthood ductopenia.

Objective To retrospectively analyze the anesthesia management of applying minimally invasive percutaneous neplrolithotripsy for treating hepatolithiasis.Methods The anaesthesia method,anesthetic effect,the time of resuscitation after anesthesia and side -effect of anesthesia in 86 patients who were underwent minimally invasive percutaneous neplrolithotripsy for hepatolithiasis were analyzed.Results All 86 patients were satisfied with anesthetic effect and completed surgery successfully.Among them,48 patients underwent operation with epidural anesthesia,31 patients with general anesthesia and 7 patients with the management of anesthesia monitoring(MAC) plus with local anesthesia.The time of resuscitation after anesthesia in general anesthesia patients was longer than those of epidural anesthesia and MAC.6 patients were delayed recovery and hypothermia after general anesthesia, 4 cases of epidural anesthesia and 1 case of MAC appeared cholecyst -heart reflection,and 2 cases of epidural anesthesia vomiting intraoperation.Conclusion The rational anesthesia method for patients underwent minimally invasive percutaneous neplrolithotripsy for treating hepatolithiasis should consider reasonably heart and lung function, maintain hemodynamics and respiratory stably,pay attention to insulation intraoperation,prevent cholecyst -heart reflection and shorten the operation time,which can reduce the side -effect of anesthesia and were favor for recovery after operation.

Objective To study the effect of MRCP for operation difficulty assessment before laparoscopic cholecystectomy. Methods 110 patients with gallbladder stone were randomly selected from March 2011 to July 2012 in our hospital,and all the patients were given LC operation after examined by MRCP. Regarded the average operation time 30 min as a criterion,when the operation time was no more than 30 min,the operation was considered as normal operation;when the operation time was more than 30 min, the operation was considered as op-eration of a certain degree of difficulty. Made a correlation analysis between LC operation difficulty and age,gender,location,volume of gall-bladder,angle of ductus cysticus, thickness of gallbladder wall, circumstances surrounding the cystic duct, length of ductus cysticus, and whether there were calculus in the cervical part of gallbladder. Results The difficulty level of LC operation is related to length of ductus cys-ticus and whether there were calculus in the cervical part of gallbladder,and there is no correlation with age,gender,location,volume of gall-bladder,angle of ductus cysticus,thickness of gallbladder wall,circumstances surrounding the cystic duct. Conclusion Routinely MRCP test in preoperative patients is of important significance to the prediction of LC operation difficulty.