Objective:To explore the mRNA expression of INSL3 gene in perinatal mice Leydig cells exposed to diethylstilbestrol(DES) in vivo and in vitro.Methods:The Leydig cells were isolated from fetal mouse testis and were cultured in DMEM/F12 medium and identified by 3 beta-hydroxysteroid dehydrogenase(3?-HSD).Reverse transcriptase-polymerase chain reaction(RT-PCR)and in situ hybridization were used for examination of the expression levels of INSL3 mRNA in primarily cultured Leydig cells incubated with DES(at the dose of 1?g/L,5?g/L and 25?g/L,respectively).Pregnant mice were exposed to DES at the dose of 1,10 or 100?g/kg body weight per day by gavage from gestation day(GD)12 to postnatal day(PND)3.The expression of INSL3 gene in neonatal mouse testis was investigated by RT-PCR.Results:Histological alterations of primarily cultured Leydig cells and neonatal mouse testis exposed to DES were observed.The expression level of INSL3 mRNA in DES treated groups including the primarily cultured Leydig cells and male mouse offspring’s testis were lower than those of the control groups.Conclusion:DES can downregulate the expression level of INSL3 mRNA in Leydig cells in vitro and in vivo.It may be a possible mechanism that DES interferes with gubernaculum development and cause cryptorchidism.

Objective To study the best method of radiation protection shielding calculation for the gamma photon produced in positron annihilation.Methods With 18F as the typical of nuclide,different methods or literature about constant and the calculation of the TVL or recommended values were adopted to calculate and analyze the shielding thickness by lead as block material.Results The shielding thickness by the dose constant was calculated by point source model (0.142 μSv· m2· MBq-1 · h-1) and TVL of lead recommended by NCRP was the biggest:at the control area the unit thickness for lead shield was 7.562 × 10-4 mm· MBq-1 for one patient.While by the dose constant recommended by AAPM TG108 (0.092 μSv·m2 ·MBq-1 ·h-1) and lead TVL recommended by IAEA was the minimum.The unit thickness for lead shield at the control area was 4.982 × 10-4 mm · MBq-1 · patient-1,with difference of 36.2％.Conclusions The dose constant and the TVL value of lead from AAPM.TG 108 are recommended for the calculation.In shielding design attention should be paid to the distance protection,the separate toilet with shielding for patients,and the separate corridors beteen doctors and patients.

Objective:To explore the changes in somatosensory evoked potential (SEP) in rats with spinal cord injury (SCI) and the effects of relieving spinal cord compression at different times on recovery and the evoked potential.Methods:Seventy Sprague-Dawley rats were randomly divided into a control group of 10 and an experimental group of 60. The experimental group was further divided into a mild injury group of 10, a moderate injury group of 40 and a severe injury group of 10. Spinal cord injuries with different severities were established in the experimental group using a self-made percussion device. The rats′ SEPs were measured before the injury, and 5 minutes, 1 hour, 6 hours, 3 days and 7 days afterward. Some of the rats receiving decompression treatment.Results:The more seriously the spinal cord was injured, the longer was the latency and the smaller was the amplitude. Both differences were statistically significant. Rats with longer compression time had significantly longer incubation periods and greater decreases in the amplitude. After relieving the compression, rats from whom it had been relieved earlier had quicker amplitude recovery. For rats under compression for 30 minutes, their amplitude was the lowest seven days later.Conclusions:For spinal cord injury, longer compression time can lead to more significant changes in the latency and amplitude of SEP, with the change in the amplitude more significant than that in the latency.

Objective To develop a modified method of adherent culturing NSCs to replace the traditional suspension culturing method.Methods Neural stem cells(NSCs)were isolated from brain of fetal mouse at 15.5-day fetal age.In culture bottle which had been pre-coated with poly-L-ornithine hydrochloride/fibronectin(PO/FN),the cells were adherently cultured and passaged.The cell morphology was observed under inverted microscope.NSCs and their differentiated cells were identified by immunofluorescence.Results NSCs obtained in this study were successfully adherently cultured and expressed specific markers.Conclusion NSCs are successfully adherently cultured in this study.As compared with the traditional suspension culturing method,adherent culturing is simpler and has lower wastage of passage.It is easy to observe cell morphology and differentiation by this method.It is also helpful to culturing and storage of NSCs.

IgG4-related disease (IgG4-RD) is a newly recognized disease entity. IgG4-RD is characterized by a single or multiple masses in one or more organs; a lymphoplasmacytic infiltrate with a high percentage of plasma cells within the lesion staining for IgG4; a peculiar pattern of fibrosis known as "storiform" fibrosis; and elevated serum IgG4 concentrations. IgG4-RD can occur in various organs, including pancreas, kidneys, lungs, retroperitoneum, and prostate gland. The head and neck involvements of IgG4-RD have been chiefly described in Mikulicz disease (MD), Küttner's tumor, orbital? inflammatory pseudotumor, and idiopathic hypertrophic pachymeningitis (IHP) previously. Recent studies reported that IgG4-RD could also involve ear, nose and throat. Here we reviewed the literatures about ear, nose and throat involvement by IgG4-RD, in order to provide some theoretical bases for the diagnosis and treatment of IgG4-RD.
Autoimmune Diseases ; physiopathology ; Ear ; physiopathology ; Fibrosis ; Humans ; Immunoglobulin G ; Nose ; physiopathology ; Pharynx ; physiopathology ; Plasma Cells ; pathology

Objective To explore the expression profile of Wnt4 in rat kidney during renal development and its effect on renal development. Methods Rats with embryonic age of 18 days (E 18 d) , 20 days (E 20 d) as well as postnatal age of 0 day (P 0 d), 1 day (P 1 d), 3 days (P 3 d), 5 days (P 5 d) and 7 days (P 7 d) were selected. Expression levels of Wnt4 in rat kidney during renal development were quantified by immunohistochemistry and Western blot in all time points. Results Immuno?histochemistry analysis showed that during E 18 d to P 7 d, Wnt4 mainly expressed in proximal tubules, ureteric bud, comma shaped bodies and S shaped bodies of nephrogenic zone;the expression in the distal tubule was weak;the expression in renal corpuscle decreased with time;Western blot analysis showed that the expression of Wnt4 in rat kidney began to decrease from E 18 d and reached bottom at P 1 d then rise again until P 7 d when it dropped again. Conclusion During renal development, Wnt4 proteins were involved in the development of the nephrogenic zone through regulating canonical Wnt/β-catenin signaling pathway, and was involved in extension of proximal tubules by inducing the non canonical Wnt/PCP signaling pathway. Expression of Wnt4 protein in rat kidney was closely related to nephron formation and development of proximal tubules.

Objective: To explore the effects of peritoneal dialysis on acute renal insufifciency and on relevant blood indicators in children with congenital heart disease (CHD) after the operation.
Methods: A total of 48 CHD patients received direct open heart surgery by cardiopulmonary bypass and suffered from post-operative acute renal insufifciency in our hospital from 2011-12 to 2014-12 were retrospectively analyzed. The patients were divided into 2 groups: Peritoneal dialysis group and Routine medication group,n=24 in each group. The differences of renal function indexes, the blood levels of electrolyte and inlfammatory factors were compared between 2 groups.
Results: Compared with Routine medication group, the patients in Peritoneal dialysis group presented decreased serum critinine, BUN and urine β2-micro globulin, 24-hour protein level,P<0.05; decreased blood K+ level and increased blood levels of Ca2+, HCO-3P<0.05; decreased plasma levels of hs-CRP, TNF-α, IL-6,P<0.05. The cure rate and mortality in Peritoneal dialysis group were better than those in Control group,P<0.05. No long term complication related to peritoneal dialysis was found.
Conclusion: Peritoneal dialysis may improve the renal function in CHD patients with post-operative acute renal insufifciency and optimize the blood levels of electrolyte and inlfammatory factors.

Objective To explore the technology of excision,and finishing on donative pancreas during combined pancreas-kidney transplantation.Methods We successfully harvested multiple abdominal organs together on 40 cases.Wide surgical exposure was obtained.Cannulas were placed for in situ cooling in portal vein and abdominal aorta,and flushed with HC-A (2000 ml) and UW (1000 ml) with the pressure being 10 cm H2O.The intestine was flushed with 0～4 ( normal saline (1000 ml) and metronidazole (200 ml),the liver,kidney,pancreas,spleen and duodenum were en bloc excised and isolated,and the pancreas and kidney were pruned.Results Excision of donative abdominal organ was successfully performed on all 40 cases.The en bloc warm ischemic time was 3.2 min (2～5 min).The cold ischemic time of pancreas was 10.6 h (8～15 h).The cold ischemic time of kidney was 8.5 h (4～16 h).Post-operation mean withdrawal-insulin time was 9.5 days (4～17 d),FFG 6.7 μmol/L (4.4～10.7 μmol/L),GHbA1c 4.4 ％ (4.1 ％～4.7 ％).Creatine was 87.2 (56～121) μmol/L one month after operation.There were 2 cases of DGF after operation,and the creatine level was returned to the normal within one month after operation.Conclusion Technology of excision,preservation and finishing on donative pancreas for combined pancreas-kidney transplantation was one of the key points for successful transplantation.

Objective Getting the robust exogenous gene expression vector under the control of porcine insulin promoter, and to lay the foundation for pancreaticβ-cells specific transgene expressing pigs.Method Using porcine insu-lin promoter ( PIP, 1500 bp of the 5′UTR from the porcine INS gene including the first exon and the first intron) to con-struct expression vector, the HindIII restriction site which connected the sequences of PIP and EGFP was designed before ATG, named PIP-HindIII-EGFP.Considering that the different location of restriction site may affect the expression efficien-cy of the transgene, we optimized the expression vector.Firstly the HindIII restriction site was deleted to realize the seam-less connection of PIP and EGFP,the vector was named PIP-EGFP.Also we mutated the 3′intron splicing acceptor site( SA) of the first intron into HindIII restriction site, named as PIP-SA( M)-EGFP.Three different EGFP expression vectors were respectively transfected MIN-6 mouse pancreatic β-cells, pig ear fibroblasts and kidney cells.The transfected cells were cultured for 48 h and harvested for RT-PCR, flow cytometry and Western blot analysis, to analyze and compare the expres-sion efficiency of vectors.Results After transfection,green fluorescence was observed only in MIN-6 mouse pancreaticβ-cells.RT-PCR analysis and product sequencing showed that the three expression vectors did have different stability with in-tron splicing.The PIP-HindIII-EGFP construct and PIP-EGFP vector produced two kinds of mRNA with the first intron spliced and no spliced, indicating the instability of intron splicing.Mutation of the PIP splice site would cause the first in-tron not spliced, while flow cytometry and Western blot displayed that the mutation induced a most efficient expression of the downstream gene.Conclusions A robust and specific β-cells expression vector has been successfully generated by mutating the intron splicing acceptor site of the porcine insulin promoter.It provides the foundation for preparation of pigs with pancreaticβ-cells specifically expressing the transgene.

Objective To summarize the clinical effect of Doty procedure on surgical treatment for patients with congenital supravalvular aortic stenosis (SVAS).Methods Between Jan.2009 and Jun.2013,12 cases of consecutive patients underwent surgical corrections of SVAS using Doty procedure.Among them,Williams' syndrome were present in 5 patients.Nine cases were male,and 3 cases were female.Mean age was(5.3 ±4.2) years(9 months to 12 years) and mean weight was (11.5 ± 5.3) kg (7.5-32.0 kg).Echocardiography and Computed Tomography confirmed the diagnosis.SVAS was discrete in 9 patients and diffuse in 3 patients.Mean peak preoperative gradient was 6.86-16.23 (11.07± 3.68) kPa,with 3 patients above 13.33 kPa.Bicuspid aortic valve was present in 2 patients,mild aortic valve stenosis in 1 patient,mild aortic regurgitation in 2 patients,1 patient with subaortic membrane,3 patients with stenosis of the left or the right pulmonary artery,1 patient with pulmonary valve stenosis,2 patients with PDA,moderate to severe mitral insufficiency in 1 patients.Surgery was performed using Doty procedure through median sternotomy with cardiopulmonary bypass.Results Mean time of cardiopulmonary bypass was (85 ± 26)min (68-129 min).Mean time of aortic clamp was (51 ± 16) min(43-68 min).There was no early death and low cardiac output syndrome.Mild ST segment change was present in 5 patients and occasional premature beat was in 3 patients.Ventilator was applied for 7 to 38 h and Cardiac Intensive Care Unit treatment was done for 1 to 4 d.The mean peak supravalvular gradient measured postoperatively was (2.18 ± 1.09) kPa.Follow-up data were available for 11 patients,ranged from 6 months to 3 years.The gradients across supravalvular were lower than 3.33 kPa,except for 2 patients with severe SVAS preoperatively.There was no late death,dizzy or palpitation during follow-up.Conclusion Doty produce for SVAS can get satisfactory clinical results.