Achromatopsia is a kind of autosomal recessive cone disorder.It occurs with nystagmus,photophobia,inability of color discrimination and severely reduced visual acuity.Five pathogenic genes had been reported to be associated with achromatopsia:cyclic nucleotide-gated (CNG)A3,CNGB3,guanine nucleotide binding protein alpha transduction active pepitide 2(GNAT2),phosphodiesterase (PDE)6C and PDE6H.They are crucial for cone phototransduction.Mutations of these genes can induce achromatopsia.Gene therapy,which can recover partial visual function,has been successfully used for the treatment of achromatopsia in animal model.Clinical features,pathogenic genes functions and mutations,the animal models and gene therapy of achromatopsia were reviewed.

Objective To detect the serum levels of carbohydrate antigen 199(CA199),carbohydrate antigen 125 (CA125)and carcino-embryonic antigen(CEA)in different population,and to explore their significances in diagnosis of pancreatic cancer.Methods The serum levels of CA199,CA125 and CEA in 56 patients with pancreatic cancer (pancreatic cancer group ), 5 6 patients with benign pancreatic disease (benign pancreatic disease group ) and 56 healthy volunteers(normal control group)were detected,then the positive rates of CA199,CA125 and CEA and the sensitivities and specificities of CA199,CEA and CA125 for the diagnosis of pancreatic cancer were compared among three groups.Results The levels of serum CA199,CA125 and CEA of the patients in pancreatic cancer group were significantly higher than those in benign pancreatic disease group and normal control group (P<0.05). The positive rates of CA199,CEA and CA125 in pancreatic cancer group were 87.50 %,71.42%,and 60.71%, respectively. The positive rates of the three tumor markers of the patients in pancreatic cancer group were obviously higher than those in other two groups (P<0.05).The sensitivity of CA199(87.50%)was the highest,while the specificity of CEA (9 1.9 6%)was the highest. The method of parallel combined testing increased the diagnostic sensitivity to 95.12% and negative predictive value to 90.47%,but its specificity and the positive predictive value were decreased. Serial combined examination increased the diagnostic specificity to 92.86% and the positive predictive value to 89.19%,but its sensitivity and negative predictive value were decreased.Conclusion Combined detection of serum levels of CA199,CA125 and CEA has higher sensitivity and specificity than single-marker detection in the diagnosis of pancreatic cancer.

ObjectiveTo study the dose and time effects of X-ray radiation on the expression of Pokemon gene and protein in human lung adenocarcinoma cell line A549.MethodsA549 cells was exposed to different doses of X-ray (2,4,6 and 8 Gy),and the expression of Pokemon mRNA and protein of the cells was detected by using Quantitative real-time PCR and western-blotting at 2,4,8,12,24,and 48 h after irradiation. 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazoliumbromidewasused to detectthe proliferation of A549 cells at 1,2,3,4,and 5 d after 2 Gy X-ray irradiation.The mock treated A549 cells were used as the control.ResultsThe expression of Pokemon mRNA trended to decrease after irradiated with 4,6 and 8 Gy in the earlier period and increased in the later period with statistical difference at the most time points (t =3.40 -154.76,P =0.000 -0.041 ).The expression of Pokemon protein trended to increase and reached the peak at 8 h after irradiated of 2,4,6 and 8 Gy with statistical difference at the most time points ( t =4.18 - 89.64,P =0.000 - 0.039).Compared with the control,the proliferation of A549 cells was significantly inhibited during 3 to 5 d after irradiation of 2 Gy ( t =2.34 - 18.19,P =0.000 -0.040).ConclusionsX-ray irradiation may increase the expression of Pokemon mRNA and protein in A549 cells,which might be correlated with radiation-resistance of A549 cells.

Objective To construct a lentiviral vector expressing small-hairpin RNA(shRNA) targeting Smad3 gene.Methods The targeting sequence of Smad3 gene which can be effectively silenced in RNA inference was confirmed in our previous study.The cDNA containing both sense and antisense Oligo DNA fragments of the targeting sequence was designed,synthesized and cloned into the pGCSIL-GFP vector.The obtained lentiviral vector containing Smad3 shRNA was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector GC-shSmad3,pHelper 1.0 and pHelper 2.0.The titer of virus was tested according to the expression level of GFP.Results PCR and DNA sequencing demonstrated that the constructed lentivirus vector GC-shSmad3 produced Smad3 shRNA.The titer of concentrated virus was 3?108 TU/ml.Conclusion The lentivirus RNAi vector targeting Smad3 is constructed successfully.

Objective To establish a co-culture system between mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) in vitro, and study the influence of SMCs on the differentiation of bone marrow MSCs (BMSCs) into SMCs in the co-culture system. Methods Density gradient centrifugation was used to separate and culture BMSCs, and collagenase digestion was employed to separate and culture bladder smooth muscle cells(BSMCs). Then the 2 types of cells were identified respectively. Then the cells were divided into 4 groups, BMSCs culture group (positive control), BSMCs cells culture group (negative control), BMSCs and BSMCs co-culture groups with contact or non-contact. The expression of ?-smooth muscle actin (?-SMA) and calponin were detected by immunofluorescence staining. Western blot analysis was employed to determine protein expression of calponin. Results In contact co-culture group, the positive rate of CD90 abd CD44 were 99.78% and 60.27% respectively in BMSCs, while only 0.21% for CD45. Western blotting showed that BMSCs expressed little calponin, but those contacting with BSMCs had an increasingly stronger expression of calponin with the elapse of contact culture time, while there was no significant change in its expression in BMSCs with contact. Immunofluorescence staining indicated that part of BMSCs were calponin positive at the 8th day after culture, but no positive cell was observed in non-contact culture group and BMSCs culture group. Conclusion The contact between BSMCs and BMSCs can promote the differentiation of BMSCs into SMCs.

Objective To explore how the different intervention measures affect the outcome of central venous catheter-related thrombosis (CRT) in children. Methods A retrospective analysis was carried out to collect the clinical data of patients with CRT from the nursing management system of the Children′s Hospital Affiliated to Zhejiang University School of Medicine which reported by each nursing unit from January 1,2015 to December 31,2015. Results Totally 108 cases were included (72 boys and 36 girls), median age of 24.5 months (ranged from 1 month to 14 years old). Nearly 42.59% (46/108) patients suffered from neurological diseases. Nearly 55.56%(60/108) CRT was detected in the first week after catheterization. Once CRT conformed, there were four kinds clinical intervention options applied. Intervention 1: thrombolytic therapy with urokinase combined anticoagulation with nadroparin calcium. Intervention 2: anticoagulant therapy only. Intervention 3: thrombolytic therapy alone. Intervention 4: no medications. The differences of effective between the four kinds of intervention were statistically significant (χ2=13.380, P=0.004). The single-factor regression analysis was done to each relevant factor. Finally the multivariate regression analysis showed four factors had impacts upon the results. The factors were as follows:gender (OR=10.400, 95%CI 1.879-57.563, P=0.007);interval (OR=1.107, 95%CI 1.035-1.184, P = 0.003), size of thrombus(OR = 1.562,95%CI 1.033-2.362,P=0.035; Intervention 2 (OR =11.757, 95% CI 2.254- 61.327, P = 0.003), intervention 4 (OR = 35.397, 95% CI 3.493-358.760, P =0.003). Conclusions The earlier and small size thrombus is more soluble. Thrombolytic therapy or combined anticoagulation is more effective. It is recommended that if no contraindications presents, thrombolytic combined with anticoagulant therapy should start early standardized treatment.

Nasopharyngeal carcinoma (NPC) poses serious health problems in Southern China and yet the molecular mechanism of the carcinogenesis remains unclear. We used modern proteomic technologies to compare the protein expression profiles between the NPC cell lines (HNE1 and CNE1 ) and an immortalized nasopharyngeal epithelial cell line NP69 to identify cancer related proteins. Cell lysates were separated by two-dimensional gel electrophoresis (2 DE ) and analyzed by PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). We discovered 15 up-regulated proteins and 18 down-regulated proteins in both HNE1 and CNE1 cell lines compared with NP69. These proteins are correlative with various functions, such as cell proliferation, apoptosis, cancer metastasis, metabolism, cytoskeleton and signal transduction. Western blotting analyses were further carried out to verify the differential expression of individual proteins. Several identified proteins in our research might be used as potential molecular markers to understand the molecular mechanism of NPC development and metastasis, and might be used as candidate targets for NPC treatments.

Objective To study the effects of muscular trumatic fluid on the biological properties of muscle-derived stem cells (MDSCs) in rats.Methods MDSCs were isolated and purified by the preplate technique,muscle injury was made for the extraction of muscular traumatic fluid.Bradford colorimetric was used to detect the protein content in the muscular traumatic fluid,and the fluid with the highest protein was used to co-culture with MDSCs.Methyl thiazolyl tetrazolium (MTT) assay and wound model of monolayer of cells were used to observe the effects of wound fluids on the proliferation and mobility of MDSCs.The expressions of α-SMA and Vimentin were tested by immunohistochemistry and Western blot technique.Results MTT assay showed that wound fluid with concentration of 10% could most significantly increase the proliferation of MDSCs.The increased expressions of α-SMA and Vimentin were found in MDSCs after cultrue with muscular traumatic fluid in vitro,and the time-dependent relation exists.Conclusions Wound environment can directly participate in the muscle regeneration by inducing the proliferation,mobility of MDSCs;on the other hand,MDSCs can differentiate into fibrotic cells under stimulation of muscular traumatic fluid.

Objective To observe the effect of home nursing based upon protocol nursing theory on liq-uid intake compliance improvement of patients undergoing hemedialysis. Methods Forty-eight HD patients of hquid intake un-compliance were randomized into the observation group and the control group with 24 cases in each group. The control group received routine nursing, the observation group was given home care instruc-tion based upon protocol nursing theory for 1 month. The two groups both received follow-up for 3 months and their liquid intake comphance were evaluated before and after intervention. The relationship between family sup-port and relative increase of body weight during hemodialysis was also evaluated. Results The IDGW relative magnitude was lower, the compliance of liquid intake was higher, the level of family support was higher in the observation group than those of the control group. There was negative correlation between the family support and the IDGW relative magnitude the IDGW relative magnitude, family support, comphance of liquid intake. Conclusions Home care nursing based upon nursing theory facilitate patients to get effect family support, increase the compliance of hquid intake and make IDGW within desirable range.

Objective To investigate the spectrum,diagnosis,time of therapy and management of the congeni-tal heart disease(CHD)in patients with Down′s syndrome(DS).Methods A retrospective report was undertaken of 96 cases in children with DS accompanied by CHD in Department of Pediatric Cardiology,Beijing Anzhen Hospital Af-filiated to Capital Medical University.Data were collected and analyzed about their clinical characteristics,and types of cardiovascular abnormalities,and the important laboratory examinations such as echocardiography and catheterization as well as the procedures of diagnosis and treatments were summarized.Then the interventions,complications and prognosis of different patients were estimated.Results (1)Single congenital heart disease was found in 33 cases (34.38%),a-mong which ventricular septal defect was the most common (14 cases,14.58%),followed by atrioventricular septal de-fect and atrial septal defect (equally,7 cases,7.29%).Multi -cardiovascular abnormalities were discovered in 63 ca-ses,and patent ductus arteriosus turned out to be the most common (42 cases,66.67%).(2)Cat-heterization was car-ried out in 18 cases of serious pulmonary arterial hypertension,and 8 cases were proved resistant pulmonary arterial hy-pertension without operation opportunity.The other 8 cases were estimated as high pulmonary arterial hypertension and medical therapy was suggested before reassessment to reduce surgical risks.(3)Operations were undertaken in 61 ca-ses,among which percutaneous interventional occlusion was performed in 7 cases and surgical interventions were per-formed in 54 patients,in which perioperation complications and death were found in 5 cases and 4 cases,respectively. Conclusions Operation interventions are practicable and most cases recovered well with systematic examinations and assessment in patients with DS and cardiovascular malformations.Early diagnosis and timely interventions are highly suggested.Also close attentions should be paid to follow -up and re -estimation after medical therapy.