Objective To explore whether Helq deletion affect the pluripotency of stem cells. Methods Helq knockout embryonic stem cells were obtained by CRISPR?Cas9 gene editing technique. Results The results of immunoflu?orescence analysis showed that the expression of Oct4 and Nanog had no obvious difference to that of the control cells. The Helq-/ - embryonic stem cells could produce viable pups by tetraploid complementation, indicating that their pluripotency was not affected. Meanwhile, we found that day 2 epiblast?like cells also were obtained through differentiation of the Helq-/ - embryonic stem cells in vitro. Immunostaining and real?time PCR analysis showed that the gene expression of Helq-/ - epiblast cells were similar to the wild type cells. Conclusions Taken together, it is proved that the genomic in?stability caused by Helq deletion does not affect the pluripotency of pluripotent stem cells.

Objective To investigate the effect of electroacupuncture on serum insulin (INS) levels and gastric antral cholecystokinin (CCK) content in rats with diabetic gastroparesis (DGP).Methods Sixty SD rats were randomized into groups A, B, C, D and E, 12 rats each. Group A is a normal control. A DGP model was made by an intraperitoneal injection of 2% streptozocin (STZ) plus an irregular high sugar and fat diet in groups B, C, D and E. group B is a model one and did not receive treatment. Group C received electroacupuncture at points Zusanli, Sanyinjiao and Liangmen; group D, electroacupuncture at the control points of Zusanli, Sanyinjiao and Liangmen. Group C, an oral gavage of metoclopramide tablet solution. Blood sugar and urine sugar were determined using a OneTouch lood glucose meter and Tes-Tape, respectively. The gastric emptying rate and the intestinal migration rate were measured by an oral gavage of phenol red. Serum INS levels and gastric antral CCK content were measured by ELISA in every group of rats before and after treatment.Results There were statistically significant differences in blood and urine sugar values between group B, C, D or E rats and group A (P<0.01) and between group C rats and group B or E (P<0.05). There was a statistically significant difference in the gastric emptying rate between group B, C, D or E rats and group A (P<0.01) and between group C rats and group B or D (P<0.01,P<0.05). There was a statistically significant difference in the intestinal migration rate between group B or D rats and group A (P<0.01) and between group C rats and group B or D (P<0.01). There were statistically significant differences in serum INS levels and gastric antral CCK content between groups B and A rats (P<0.05,P<0.01). There was a statistically significant difference in gastric antral CCK content between groups D and A rats (P<0.01). There were statistically significant differences in serum INS levels and gastric antral CCK content between groups C or E rats and group B (P<0.05,P<0.01). There was a statistically significant difference in gastric antral CCK content between groups C and D rats (P<0.05).Conclusions Electroacupuncture can markedly improve gastrointestinal functions and promote gastric emptying in DGP rats. Its therapeutic effect may be related to electroacupuncture raising serum INS levels and reducing gastric antral CCK content.

Objective To primarily study the possible biological significance of yon Willebrand factor (vWF) expression at the fracture site in the mechanism of fracture healing. Methods A total of 28 male SD rats were selected to set up femoral fracture models. Then, tissue samples from fracture site were randomly taken at days 2, 3, 5, 7, 10, 14 and 21 after establishment of the models, with 4 rats at each time point, vWF expression at fracture site and the relationship of vWF expression with osteoblasts and chondroblasts were observed by using computer image analysis system. The biological significance of vWF expression at fracture site in mechanism of fracture healing were analysed based on the results of vWF expression of in vitro cultured rat osteoblasts. Results The expression of vWF at every time point was positive and distributed disproportionally irregular at bone trabecula and soft tissue of the fracture site. vWF expressed positively in the extracellular matrix around the chondroblasts. Conclusions During the course of fracture healing, there may be vWF-platelet activation pathway that can regulate differentiation and proliferation of local osteoblasts and chondrocytes and activate platelets secreting growth factors such as PDGF, IGF-1, EGF and TGF-a, as creates a sound microenvironment for fracture healing.

Objective To detect epidermal growth factor receptor (EGFR) 19, 21 exon gene mutation and gene copy number in lung adenocarcinoma tissue, and to analyze the relationship of EGFR 19, 21 mutation with copies number. MethodsEGFR mutations and gene copy number in the tissue samples embedded by paraffin and fixed by for marlin from 58 cases of lung adenocarcinoma were detected by RT-PCR and FISH. The statistical data were analyzed by chi-square test.ResultsOf 58 cases, the overall single mutation rate of EGFR exon 19, 21 was 43.1% (25/58), and 2 cases contained both types of the mutation.The overall FISH positive rate of EGFR was 51.7 % (30/58), including 8 positive amplification and 22 highly ploidy amplification. The testing results showed that there had no statistically differences in FISH positive rates of EGFR mutation among different differentiation lung adenocarcinoma tissues(P ＞0.05), and the FISH positive rates of EGFR mutation in poorly differentiated cancer were lower than those in moderatedly differentiated and well-differentiated cancer (P ＜0.05). EGFR mutation was closely related to EGFR gene copies (P ＜0.01). ConclusionThere are high EGFR mutation frequencies and FISH positive rates in lung adenocarcinoma tissue; Combined detection of EGFR mutation and gene copy number may provide a better approach in selecting patients who may benefit from anti-EGFR target therapy.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Oximetry ; Pneumoconiosis ; blood ; complications ; Prognosis ; Respiratory Insufficiency ; blood ; etiology

Objective: To explore the action mechanism of electroacupuncture (EA) in improving the gastric motility of rats with diabetic gastroparesis (DGP). Methods: Forty-eight healthy Sprague-Dawley (SD) rats were randomly assigned into four groups: a normal group (group A), a model group (group B), a group of EA at acupoints (group C), and a group of EA at non-acupoints (group D), 12 rats in each group. The animal model of DGP was established by intraperitoneal injection of streptozotocin (STZ) plus high glucose and fat diet. The blood glucose, urine glucose and gastric emptying rate (GER) were observed. The content of insulin (INS) in serum and ghrelin in gastric antrum tissue were detected by enzyme linked immunoassay (ELISA). The expression of growth hormone secretagogue receptor mRNA (GHSR mRNA) in gastric antrum tissue was detected by real-time fluorescent quantitative polymerase chain reaction (PCR). Results:Compared with group A, blood glucose and urine glucose increased significantly (P<0.01), GER, content of serum insulin, the content of ghrelin and expression of GHSR mRNA in gastric antrum tissue decreased significantly (P<0.05 orP<0.01) in group B. Compared with group B, blood glucose and urine glucose decreased significantly (P<0.05), GER, the content of insulin in serum, the content of ghrelin and expression of GHSR mRNA in gastric antrum tissue increased significantly (P<0.05 orP<0.01) in group C. Conclusion: EA at acupoints can down-regulate the content of blood and urine sugar, and promote gastric emptying, which is possibly related to the regulation of serum insulin, and the expressions of ghrelin and GHSR mRNA in gastric antrum.

Objective To analyze human leucocyte antigen (HLA)-A0201 restricted antigen-specific cytotoxic lymphocytes (CTL), and to investigate the difference of T cell response to specific antigen epitopes between patients with acute phase of acute hepatitis B and active phase of chronic hepatitis B. Methods Peripheral blood mononuclear cells (PBMC) from 5 patients with acute phase of acute hepatitis B and 6 patients with active phase of chronic hepatitis B were isolated. The numbers and functions of CD8+ T-lymphocyte epitope peptide specific CTL were detected using enzyme-linked immunosorbent spot (ELISPOT) assay, and the 3 peptides were from HBV polymerase region (Pol575-583), envelope region (Env348-357) and core region (Core18-27), respectively. The data were analyzed using t test. Results The spot formation cell counts (SFC) of Pol575-583, Env348-357 and Core18-27 stimulations in patients with acute phase of acute hepatitis B were 110±13, 165±17 and 185±20, respectively; and those in patients with active phase of chronic hepatitis B were 22±4, 23±5 and 30±5, respectively; the differences were all significant (t=10.9, 15.2 and 8.0, respectively, all P<0.05). The CTL responses to the three peptides in patients with acute phase of acute hepatitis B were Pol575-5830.05). The SFC were increased upon non-antigen specific HLA-A2404 restricted epitope (Core117-125), but the difference was not significant compared with negative control group (P>0.05). Conclusions Hepatitis B virus-specific CTL responses in patients with acute hepatitis B are significantly higher than those in patients with chronic hepatitis B. The number and function of polyclonal CTL are both impaired in patients with chronic hepatitis B.

Objective: To determine possible relations between early adiposity rebound and adolescent development. Methods: Prospective children cohort from 2 kindergartens selected through clustering convenience sampling method in Anhui Province was established since Sep. 2010. Participants were classified as Cohort 1 (2010), Cohort 2 (2011) and Cohort 3 (2012) according to the recruiting year. Till Sep. 2015, a toal of 802 girls were included in this study, and received follow-up till primary school. During kindergarten period, physical examination was carried out every 3 months, 8 times in total. In primary school, physical examination was carried out annually; till Sep.2015, the cohort 1, 2, 3 took physical examination for 12, 11, 10 times, respectively. Information on household economic status and child physical activity was acquired through parents questionnaire survey, and breast development were assessed through visual inspection and palpation. Adiposity rebound was determined according to Rolland-Cachera's method for each girl. Differences between early adiposity rebound and normal adiposity rebound groups were compared by using t test and χ² test. Multivariate regression analysis was applied to explore the association between early adiposity rebound and breast development. Results: The average age of participants was (8.90±0.87) years old and the BMI was (17.48±2.70) kg/m². The average age at adiposity rebound was (6.16±0.90) years old and the BMI was (15.33±1.82) kg/m². Premature breast development was found significantly higher in girls in early adiposity rebound group (27.8%, 54/802) than it in normal adiposity rebound group (13.7%) (P<0.001). After current adiposity, age, household economic status, childhood physical activity adjusted, the OR of premature beast development in early adiposity rebound group was 2.41(95%CI: 1.41-4.12). Conclusion Early adiposity rebound increases the risk of premature puberty in girls.

Aim To investigate the inhibition role and mechanism of adipose derived mesenchymal stem cell(ADMSC)exosomes(Exo)on adverse ventricular remodeling after myocardial infarction(MI).Methods The chan-ges of autophagy and inflammasomes phenotype of cardiac fibroblasts after H2O2 treatment were observed.MI rats were in-jected with an equal volume of normal saline,adipose derived mesenchymal stem cell exosomes(MSC-Exo)or fibroblast exosomes(MEF-Exo)via a tail vein.The expression of autophagy related 16 like protein 1(ATG16L1),autophagy re-lated protein 7(ATG7)and NOD-like receptor protein 3(NLRP3),inflammatory response,the degree of myocardial fi-brosis,and the cardiac function were observed in different groups.Results After treatment with H2O2 on cardiac fi-broblasts,the expressions of ATG16L1 and ATG7 were significantly decreased(P＜0.001),NLRP3 was significantly in-creased(P＜0.001),and the levels of inflammatory cytokines interleukin-1β(IL-1β)and IL-18 were significantly elevated(P＜0.001).After MI rats were intervened with MSC-Exo,the expressions of autophagy related proteins ATG16L1 and ATG7 were significantly up-regulated(P＜0.001),NLRP3 was significantly down-regulated(P＜0.001),serum IL-1β and IL-18 levels were significantly decreased(P＜0.001),fibrosis-related proteins collagen Ⅰ and Ⅲ were significantly reduced(P＜0.001),myocardial fibrosis was significantly relieved(P＜0.001),and cardiac function was sig-nificantly improved(P＜0.001).Conclusion Adipose derived MSC-Exo play a role in inhibiting adverse ventricular remodeling after MI by regulating the balance of autophagy and NLRP3 inflammasomes.