Objective To analyze the clinical features and pathogenic gene mutation in a Chinese family with cerebro-oculo-facio-skeletal(COFS)syndrome,in order to summarize the relationship between phenotype and genotype.Methods The clinical data of the proband and his family members were collected.Genomic DNA from the proband and his parents were extracted by using standard procedures from the peripheral blood leukocytes.Next-generation sequencing was used to detect gene mutation in the patient with COFS syndrome.Sanger sequencing was applied to confirm the results.Results The proband,male,1 year and 3 months old,presented with microcephaly nystagmus,large ears,prominent nose,high arched palate,overhanging upper lip,micrognathia,widely set nipples,flexion contractures(especially involving the elbows and knees),failure to thrive,developmental retardation and feeding difficulty.His parents were normal phenotype.Two different heterozygous mutations c.1843G>T(P.G615W)and c.1996 C> T(P.R666W)were identified in the ERCC2 gene.The proband's father had the heterozygous mutation c.1843G>T(P.G615W)and his mother had the heterozygous mutation c.1996 C> T(P.R666W).Meanwhile,this heterozygous mutation c.1996 C> T(P.R666W)had been reported as a pathogenic gene mutation.Conclusions COFS syndrome is characterized by microcephaly,prominent nose,arthrogryposis and severe developmental delay.This is the first report on COFS syndrome patient in the mainland of China.The pathogenic gene mutations and gene status were identified through genetic studies.The result has laid the foundation for accurate genetic counseling and further prenatal diagnosis.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.

Objective: To investigate the current status of malaria rapid diagnostic test (RDT) strips application and malaria laboratory technicians' evaluation about them at primary healthcare provider level in Jiangsu Province. Methods: From November to December 2016, 878 medical institutions and 118 CDCs of city, county and township/community level in Jiangsu Province were selected as study samples using stratified random sampling method. Self-designed questionnaire was distributed to investigate the institution's malaria work task, RDT strips application and evaluation status in 2015. We also investigated the socio-demographic information and collected the RDT strips evaluation score from the malaria laboratory technicians selected from the institutions investigated (one technician from each institution). Rank sum test was performed to compare the RDT strips evaluation scores between medical institutions and CDCs, and among different medical institutions and CDCs. Results: In 2015, 405 cases of malaria were reported, 362 200 person-time of malaria blood testing task was conducted, and 100 000 RDT strips were procured and provided for healthcare providers in Jiangsu province for free. Of the 996 healthcare institutions investigated, 628 used RDT strips in the year 2015 and the median (P₂₅, P₇₅) of RDT strips volume used in these institutions was 10 (2, 25). The volume of RDT strips used in CDCs (15 (5, 52)) was significantly higher than that in medical institutions (10 (2, 25), (Z=3.42, P=0.001)). The investigated CDCs gave higher score on RDT strips' testing time per operation (10 (8.5, 10)) than medical institutions (9(8, 10), (Z=-2.20, P=0.028)). The employers of 614 investigated malaria laboratory technicians used RDT strips in 2015. The median of the scores given by CDC malaria laboratory technicians for RDT strips in terms of testing time per operation, testing operation and results judgement difficulties were 10 (9, 10), 10 (9, 10) and 10 (9, 10), respectively, which were significantly higher than those from technicians of medical institutions (9 (8, 10), 9 (8, 10), 9 (8, 10), (Z values were -2.55, -2.97 and -2.96, respectively; P values were all less than 0.05)). Conclusion RDT strips had been widely performed in health institutions in Jiangsu Province. The amount of RDT strips used in CDCs was significantly higher than that in medical institutions. Primary-level institutions and malaria laboratory technicians generally recognized RDT strips' advantage for application in terms of testing time and operational procedure. CDCs and malaria laboratory technicians from them gave higher regards on RDT strips in terms of testing time per operation, testing operation and results judgement difficulties compared with that of medical institutions.

Objective To investigate the difference in intestinal flora among patients with esophageal squamous cell carcinoma and normal population and to provide a basis for the early diagnosis of esophageal squamous cell carcinoma as a marker. Methods DNA was extracted from biopsy tissue samples of 30 patients with esophageal squamous cell carcinoma (observation group) and 25 healthy people (control group) by microbial amplification sequencing. The integrity and quality of DNA were detected. The composition and abundance of intestinal flora in the samples of the two groups were determined. Results A great similarity in beta diversity was found between the two groups, but some differences were also observed. The relative abundance of Proteobacteria and Verrucomicrobia in the observation group was higher than that in the control group (P<0.05). The relative abundance of Megamonas in the observation group was lower than that in the control group (P=0.025). Conclusion Strengthening the study on the changes in intestinal flora among patients with esophageal squamous cell carcinoma may be of great significance for its prevention and treatment.

Objective To explore the safety and efficacy of intrathecal administration of adipose stem cells de-rived from bioactive secretome (ASCBS)in treatment of whiter matter injury (WMI)in the preterm infants. Methods Sixty - three cases of WMI were recruited according to the uniform standards from multiple medical centers and they were divided into 3 gestational age (GA)subgroups,which were 21 cases in group A (GA 24 - 28 + 6 ),20 cases in group B (GA 29 - 32 + 6 ),and 22 cases in group C (GA 33 - 36 + 6 ). The patients were randomly divided into treatment groups and control groups by tossing coins. The treatment groups received lumbar puncture followed with ASCBS intra-thecal injection once daily for 3 consecutive days. Follow - up study included Neonatal Behavioral Neurological Assess-ment (NBNA)at term - equivalent age and neurodevelopment at corrected age of 6 - month. Neurodevelopment was assessed by using the Bayley Scales of Infant Development and Peabody Developmental Motor Scale. The survival rates, NBNA scores,mental development index (MDI),psychomotor develop index (PDI),total motor development quotient, gross motor development quotient and fine motor development among each subgroup were compared. Results Sixty -three cases were recruited,including 31 in the treatment group and 32 in the control group. Only 1 case in the treatment groups lost in the follow - up. No clinical side effects were found in the treatment groups. There was no significant diffe-rence in the survival rate and complication in the preterms in all subgroups of the treatment group and control group (all P > 0. 05). The gross and total motor development quotient in the treatment group A was higher than that in the control group A(gross motor development quotient:98. 330 ± 6. 282 in treatment group A,90. 330 ± 3. 777 in control group A, P = 0. 040;total motor development quotient:97. 330 ± 4. 803 in treatment group A,91. 000 ± 4. 472 in control group A,P = 0. 023). The rest findings showed no significant difference between groups. Conclusion The treatment of WMI in preterm infants with ASCBS is safe and can promote the motor development of preterm infants with GA in 24 - 28 weeks.

Objective : To investigate the effects of 3 types of trihalomethanes ( THMs) , the by⁃products in treatment of drinking water disinfection , on the development and reproductive capacity of germ cells of Caenorhabditis elegans. Methods : Tribromomethane ( TBM) , trichloromethane ( TCM) and triiodomethane ( TIM) , 3 types of THMs were selected and wild type Caenorhabditis elegans was used as a model to detect germ cell apoptosis , oocyte number and the final brood size after exposure to different concentrations of THMs. Results : The results of germ cell apoptosis showed that the exposure to 50 μmol/L TCM and TIM caused elevated levels of germ cell apoptosis in a dose⁃dependent manner. Oocyte counting results showed that exposure to 50 μmol/L all three THMs significantly reduced the number of oocytes. The brood size results showed that only 50 μmol/L TIM exposure significantly de⁃ creased the brood size of nematodes. Conclusion THMs showed negative impacts on the reproductive system of Caenorhabditis elegans , and the toxicity is related to the halogenated elements of THMs.