BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are few in bone marrow, and they are easily mingled with other cells, especially red blood cells. Therefore, intervention needs to be depleted in the process of isolation of red blood cells so as to obtain highlypurified BMSCs as many as possible.OBJECTIVE: To isolate and culture BMSCs of rats with red blood cell lysis method, and perform biological identification.DESIGN: Observation and controlled trial.SETTING: Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts.MATERIALS: Fifty 30-day-old male SD rats, weighing about 100 g, of SPF degree, were purchased from Beijing Experimental Animal Center (License No. SCXK (Jing) 2002-0003) and involved in this trial. DAB condensed chromogen (hydrogen dioxide included, Zhongshan Company), rabbit anti-ret polyclonal antibody (Boster Co.,Ltd., Wuhan), fetal calf serum (PAA, Austria) and LG-DMEM medium (Sigma Company, USA) were used in this trial.METHODS: This trial was carried out in the Laboratory of Aerospace Cell Molecular Biology, Scientific Research Training Center for Chinese Astronauts during September 2004 to September 2005. The rats were sacrificed by dislocation to expose bone marrow cavity. Cell suspension was collected. BMSCs were isolated and cultured primarily by whole bonemarrow culture method. ① Red blood cells lysis test: A, B, C and D 4 tubes were chosen and filled with 0.5 mL bonemarrow rinse solution which was filtered and fully beat upon. Then, red blood cell lysis buffer of 2 mL, ammonium chloride of 2 mL, phosphate buffer normal saline of 2 mL and 0.04 volume fraction acetic acid of 0.5 mL were correspondingly added into the 4 tubes. In each tube, absorbance and hemoglobin concentration were measured and cell growth was observed. ② Observation of growth curve, doubling time and surface marker molecule expression of BMSCs: Based on the formula, population doubling time (TD) =t[lg2/(lgNt-lgN0)] (NO and Nt represented the cell number after inoculation and t hours after culture ,respectively), cell population doubling time was calculated and traced and growth curve of the 2nd, 4th and 6th generations of BMSCs were analyzed; The proliferation of BMSCs was measured by methylene blue staining method; Surface marker molecule expression of BMSCs was detected with immunocytochemical staining.MAIN OUTCOME MEASURES: ①Observation of isolation and culture of bone marrowmesenchymal stems of rats. ②Effect of different methods on the lysis of red blood cells and the growth of BMSCs. ③ The growth curve and cell doubling time of the 2nd, 4th and 6th generations of cells. ④Surface marker molecule expression of BMSCs of rats.RESULTS: ① Results of isolation and culture of BMSCs of rats: After 48-hour primary culture, most of the cells had adhered to the wall, and 72 hours later, division growth of the adherent cells presented. Seven to eight days later, cell colonies formed obviously, and then increased quickly, expanded incessantly and fused with each other. On 14 to 16 days, cell clones grew densely. Immediately generative cells presented ball-shape, subsided and adhered the wall verysoon. Some few round cells suspended. Adherent cells distributed evenly and proliferated quickly within 3 to 5 days. Although cell morphology of generative cells did not change after passage, cell proliferation was speeded up obviously and cells covered the bottom of the whole bottom on about 6 days. ② Effect of different methods on red blood cell lysis and the growth of BMSCs: hemoglobin concentration in the red blood cell lysate-treated group, ammonium chloride-treated group and 4％ acetic acid-treated group was significantly higher than that in the phosphate buffer normal saline-treated group, with significant difference (P ＜ 0.01). ③ Observation of growth curve of different generations of BMSCs: The growth curves of the 2nd, 4th and 6th generations of the cells were basically the same: latent period about 1 to 2 days, then logarithmic growth phase, peak on the 5th day and finally plateau phase (about on the 5th to 7th days). The latent period of the 6th generation of BMSCs was not obvious and the population doubling time of BMSCs was about 34 hours. ④ldentification of immunophenotype of different generations of BMSCs: Both CD44 and CD106 staining of each generation of cells were positive, presenting brown granule sediments, and CD34 staining was negative.CONCLUSION: Inoculation of red blood cells lysate-treated bone marrow rinse solution can boost the adherent rate of BMSCs, and does not influence its post-adherent growth, so it is a feasible separation method. Cell surface marker staining confirms that thecells isolated in this study are BMSCs.

To investigate the high-risk factors for newborn hearing loss and to provide information for preventing the development of hearing loss and delaying its progression, from May 2003 to June 2006, neonates who failed to pass the universal newborn hearing screening (UNHS) were referred to Jinan Newborn Hearing Screening and Rehabilitation Center from 7 newborn hearing screening centers in seven cities of Shandong province. One-to-one pair-matched case-control method was employed for statistical analysis of the basic features of definitely identified cases. High-risk factors relating to the bilateral hearing loss were evaluated by univariate and multivariate Logistic regression analysis. Our results revealed that 721 transferred newborns who didn't pass the hearing screening received audiological and medical evaluation and 367 were confirmed to have hearing loss. Of them, 177 neonates with hearing loss who met the matching requirements were included in the study as subjects. Univariate analysis showed that high-risk factors related to hearing loss incuded age of father, education backgrounds of parents, parity, birth weight, gestational weeks, craniofacial deformity, history of receiving treatment in neonatal intensive care unit (NICU), neonatal disease, family history of otopathy and family history of congenital hearing loss. Multivariate Logistic regression analysis revealed that 4 independent risk factors were related to bilateral hearing loss, including parity (OR=16.285, 95% CI 3.379-78.481), neonatal disease (OR=34.968, 95% CI 2.720-449.534), family history of congenital hearing loss (OR=69.488, 95% CI 4.417-1093.300) and birth weight (OR=0.241, 95% CI 0.090-0.648). It is concluded that parity, neonatal disease and family history of hearing loss are the promoting factors of bilateral hearing loss in neonates and appropriate intervention measures should be taken to deal with the risk factors.

Objective To establish a carrying system for space cellular experiment suitable for astronaut to carry out cellular experiments on Shenzhou-6 mission.Methods The cell carrying sample bag,sample box and sample box integrated package were designed.Primary cardiomyocytes and osteoblasts culture and ground model experiment in the simulated environment of space cabin were performed.With man-tended,the cellular experiment was carried out on the orbit.Results After 5 d space flight,the returned cell samples were analyzed.The results demonstrated that the system was of good safety,reliability and applicability,as well as satisfied the demands of analyzed samples.Conclusion After Shenzhou-6 space flight,it is showed that this system fits for small loading,multi-cells and man-tended carrying mission,and can satisfy the demand of the first man-tended space cellular experiments carried out on the Shenzhou-6 spacecraft.

AMP-Activated Protein Kinases ; metabolism ; Adiponectin ; metabolism ; Fatty Liver, Alcoholic ; pathology ; Humans ; Signal Transduction ; Sirtuin 1 ; metabolism

To investigate the high-risk factors for newborn heating loss and to provide information for preventing the development of hearing loss and delaying its progression, from May 2003 to June 2006, neonates who failed to pass the universal newborn hearing screening (UNHS) were referred to Jinan Newborn Hearing Screening and Rehabilitation Center from 7 newborn heating screening centers in seven cities of Shandong province. One-to-one pair-matched case-control method was employed for statistical analysis of the basic features of definitely identified cases. High-risk factors relating to the bilateral hearing loss were evaluated by univariate and multivariate Logistic regression analysis. Our results revealed that 721 transferred newborns who didn't pass the heating screening received audiological and medical evaluation and 367 were confirmed to have heating loss. Of them,177 neonates with heating loss who met the matching requirements were included in the study as subjects. Univariate analysis showed that high-risk factors related to hearing loss incuded age of father, education backgrounds of parents, parity, birth weight, gestational weeks, craniofacial deformity,history of receiving treatment in neonatal intensive care unit (NICU), neonatal disease, family history of otopathy and family history of congenital hearing loss. Multivariate Logistic regression analysis revealed that 4 independent risk factors were related to bilateral hearing loss, including parity (OR=16.285, 95% CI 3.379-78.481), neonatal disease (OR=34.968, 95% CI 2.720-449.534),family history of congenital hearing loss (OR=69.488, 95% CI 4.417-1093.300) and birth weight (OR=0.241, 95% CI 0.090-0.648). It is concluded that parity, neonatal disease and family history of hearing loss are the promoting factors of bilateral hearing loss in neonates and appropriate intervention measures should be taken to deal with the risk factors.

OBJECTIVEIn this study, we employed newborn hearing screening and gene screening concurrently to explore the hearing loss associated with mutations in the city of Jinan.

METHODSA total of 3 288 newborns born between March 2013 and December 2013 in Jinan Maternity and Child Care Hospital received hearing concurrent genetic screening. Transiently evoked otoacoustic emissions (TEOAE) was used in rooming-in newborns, while TEOAE and auto auditory brainstem response (AABR) was used in infants in neonatal intensive care unit (NICU). Two drops of heel blood were harvested with filter paper. Nine mutations [GJB2 (235delC, 35delG, 299delAT, 176del16), SLC26A4 (IVS7-2A>G,2168 A>G), GJB3 (538 C>T), 12SrRNA (1555 A>G, 1494C>T)] of 4 frequent genes associated with Chinese hearing loss were determined by gene chip in these dried blood samples.

RESULTSAmong 3 288 newborns, 363 cases failed to pass the hearing screening, and 36 cases of these 363 newborns carried mutations, with a carrier rate of 9.91%. 2 925 cases passed the hearing screening, of which 113 carried mutations, with a carrier rate of 3.86%. There was a significantly statistic difference (χ2=8.67, P=0.000) in carrier rate between two groups. 149 (4.53%) infants were detected to carry at least one mutation allele,among which 113 cases passed the hearing screening and 36 cases failed. Seven cases were diagnosed to have hearing loss. Homozygous GJB2 mutation was detected in 2 cases, compound heterozygous GJB2 mutation was detected in 1 case, and heterozygous GJB2 mutation in 88 cases. There were 91 cases carried GJB2 mutations totally, with a total rate of 2.76%. There were 40 cases were detected to carry heterozygous SLC26A4 mutation, with a carrier rate of 1.22%. Nine cases had heterozygous GJB3 mutation, with a carrier rate of 0.27%. Six cases had homogeneous mitochondria 12SrRNA mutation, and 1 had heterogeneous mutations. There were 7 cases totally, with a total rate of 0.21%. 142 infants with gene mutation should be follow-up.

CONCLUSIONA follow-up system in infants, passed hearing screening,with single heterozygous mutation and mutations associated with drug-induced hearing loss, can help to detect infants with hearing defects early and effectively prevent late-onset hearing impairment.

Alleles ; Asian Continental Ancestry Group ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Evoked Potentials, Auditory, Brain Stem ; Genetic Testing ; Hearing Loss ; diagnosis ; Hearing Tests ; Heterozygote ; Homozygote ; Humans ; Infant, Newborn ; Membrane Transport Proteins ; genetics ; Mutation ; Neonatal Screening ; RNA, Ribosomal ; genetics

Objective:To investigate the clinical characteristics of carotid webs and prevention measurements of cerebral ischemic stroke caused by carotid webs.Methods:The clinical data of three patients with carotid webs admitted to our hospital from October 2017 to January 2019 were retrospectively studied, and publications (208 patients with carotid webs) reported from January 1, 2014 to June 30, 2019 in PubMed, Embase, Wanfang and CNKI databases were collected. The demographic and clinical characteristics of carotid webs and secondary prevention measurements of ischemic stroke caused by carotid webs were analyzed.Results:A total of 211 patients with carotid webs were enrolled, including 68 male (32.2%) and 143 female (67.8%), with an median age of 48 years. Prevalence of risk factors for cerebrovascular disease reported in 148 patients was as follows: hypertension ( n=43, 29.1%), dyslipidemia ( n=19, 12.8%), diabetes mellitus ( n=14, 9.5%), and smoking ( n=12, 8.1%); 89.8% of carotid webs(44/49) caused mild stenosis of carotid artery (stenosis degree<50%). In secondary stroke prevention, the recurrence rate of stroke patients treated with anticoagulation plus antiplatelet was obviously lower than that of patients treated with antiplatelet alone (11.1% vs. 49.3%). No ischemic stroke or surgical complications were noted in 23 patients underwent carotid endarterectomy and 54 patients underwent carotid stent implantation during the mean follow-up period of 12 months (one-60 months) and 11 months (3-144 months), respectively. Conclusions:Carotid web may be a vital risk factor for cryptogenic stroke. Anticoagulation plus antiplatelet is superior to antiplatelet alone in preventing stroke recurrence. Carotid endarterectomy and carotid stent implantation may be the perfect choices for carotid web patients with high risk of stroke recurrence.

Objective: To evalate the effectiveness and suitability of a wearable health monitoring device for community-based management of hypertension. Methods: In December 2015, 400 patients with hypertension were enrolled from Beijing, Chaoyang. Subjects were divided into an experimental group (220 cases) and control group (180 cases), and baseline data were collected. The control group received follow-up with general planning while the experimental group received wearable health devices. Follow-up was performed three times using a questionnaire (April, August, and December 2016), and medical staff provided feedback and guidance. The experimental group was also classified according to risk factors and intervention measures were individually designed, and included monitor and medication compliance, self-management ability, and social support. Communication between patients and medical staff was recorded to form a case system. Evaluation indexes included accuracy and reliability, blood pressure management efficacy, behavior intervention efficacy, satisfaction, and disease burden. A t-test, non-parametric test, and chi-square test were used to compare the experimental and control groups before and after intervention. Results: At 1-year follow-up, after correcting for differences in baseline information between the two groups, statistically significant differences in numerical indexes were observed for number of visits within 1 month [1(1) vs. 1(1), Z=5.42], payment within 1 month [85(141) yuan vs. 40(70) yuan, Z=-2.66], visiting time [20(20) min vs. 20(15) min, Z=-2.82], exercise times [4.79(2.24) times/week vs. 4.09(2.00) times/week, Z=9.27], medication compliance score [7.33(5.77) vs. 8.70(5.24), Z=6.86], satisfaction [9.27(0.08) vs. 8.88(0.10), Z=11.77], diastolic pressure [(78.93±0.56) mmHg vs. (81.32±0.61)mmHg, F=8.70] (1 mmHg=0.133 kPa), and body mass index [(25.55±0.27) kg/m² vs. (27.74±0.43) kg/m², F=-2.24]. In addition, classification indexes adjusted for normalized blood pressure and habitus were different between experimental and control groups (χ²=3.89, 8.38, P≤0.05). The equipment worked well, with performance rates of over 90% (90.9%, 97.3%, and 92.7%). Conclusion The wearable health monitoring equipment showed good stability and reliability, and was able to effectively support health management in patients with hypertension in the community. At the same time, the equipment can improve healthy lifestyle compliance and awareness or self-management of blood pressure. In this manner, the burden of disease is reduced and the quality of life is improved.