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 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.

Objective To investigate the application and clinical value of donation after citizens death (DCD) in salvage liver transplantation (SLT).Method The clinical data of 12 recipients who underwent SLT from DCD of 12 donors at the Guangzhou General Hospital of Guangzhou Military Area from October 2010 to December 2013 were retrospectively analyzed.Of the donors,there was one case of type of China-Ⅰ,2 cases of the type of China-Ⅱ and 9 cases of the type of China-Ⅲ.Extracorporeal membrane oxygenation (ECMO) was applied to donation after brain and cardiac death to avoid warm ischemia.Of the recipients,6 met the Milan criteria,3 UCSF(The University of California,San Francisco) criteria and 3 Hangzhou criteria respectively.Orthotopic liver transplantation was performed on recipients.Eleven recipients received end-to-end anastomosis of the bile duct and followed up regularly.Result Liver transplantation was successfully performed on all recipients.No mortality during operation,no recovering delay and non-function of the transplanted liver occurred.Postoperative complications occurred in 4 cases,2 patients died,and 2 patients recovered and discharged after the second surgery.Ten recipients had a long-term survival.Two cases of hepatocellular carcinoma (HCC) recurrence were still alive after active treatment.The longest survival time was 42 months.All of them had no long-term complications such as biliary stricture.Conclusion The role and status of SLT in the treatment of HCC has become increasingly,and there will be more and more use of DCD donor in SLT.To ensure the DCD donors safely and effectively applied in SLT,we should grasp the characteristics of SLT,choose the appropriate DCD donor according to recipients,and rationally use the ECMO to protect the graft quality.

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 risk factors of electrical storm(ES) in patients with acute myocardial infarction (AMI) during perioperative period of direct percutaneous coronary intervention(PCI).Methods Forty-one AMI patients had been treated with direct PCI.The patients with perioperative ES were included in ES group and those without perioperative ES were included in conntrol group.ES was defined as the occurrence of spontaneous ventricular tachycardia or venicular fibrillation was twice or more within 24 h and unable to stop by itself and emergency treatment was needed.The difference of the clinical data between two groups were compared.Results There were 7 in 41 patients with direct PCI who had ES,the incidence was 17.07％,and 34 cases didn't have ES.Systolic pressure,diastolic pressure,white cell count,blood glucose,international normalized ratio and time duration from chest pain onset to direct PCI between two groups had no significant differences (P ＞0.05).Age,CK-MB,cardiac troponin I,the diameter of infarctrelated arleries(IRA ),incidence of reperfusion arrhythmia and mortality of ES group were all obviously higher than those of control group (P ＜ 0.05 or ＜ 0.01 ).The incidence of ES in patients whose IRA was left main artery or occlusion of middle section of two main coronary arteries,right coronary artery,left anterior descending branch and left circumflex artery was 66.67％(2/3),18.75％(3/16),11.76％(2/17) and O, respectively.Conclusions Perioperative ES during direct PCI most commonly occurrs in AMI patients with left main artery or occlusion of middle section of two main coronary artery.The diameter of IRA,TIMI flow classification after the patency of IRA and recanalization arrhythmia are the main risk factors of the occurrence of perioperative ES.

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.

ObjectiveTo analyze the etiologies and diagnosis of jaundice after liver transplantation during mid later stage and summarize the experience of clinical practice.Methods822 liver transplantation patients were collected from the Department of Internal Medicine, Organ Transplantation Institute in General Hospital of Chinese People's Armed Police Forces between June 2005 and Dec.2008.The clinical data of these cases were analyzed retrospectively.ResultsAmongst 822 patients,129 experienced jaundice after liver transplantation and the occurrence of jaundice ranged from 43 days to 39 months postoperation.Total bilirubin levels were from 27.4 to 503 μmol/L.The etiologies of jaundice and its percentage were as follow:59 cases of biliary complications (45.7％),36 of rejection (27.8％ ),11 cases of virus infection (8.5％),5 cases of drug-induced hepatic injury (3.9％),4 cases of tumor recurrence (3.1％),4 cases of Gilbert syndrome (3.1％),3.cases of dysfunction of papillary muscle (2.3％),2 cases of vascellum complication (1.6％),2 cases of radiation hepatitis (1.6％),1 case of hepatapostema (0.8％),2 cases of unknown reasons (1.6％),respectively.The jaundice of most patients (93％) got released while small part of patients (7％)failed to treatment,of whom 5 cases received liver re-transplantation and 4 cases died of disease progression.ConclusionThe etiologies of jaundice after liver transplantation during mid-later stage are diversiform and complex.And the etiological diagnosis is the premise to treat effectively.