Background Retinal hypoxia is one of primary causes of retinal neovascularization,and its mechanism is research hot topic.Studies showed that hypoxia stimulates the activation of many trancripation factors and vascular endothelial cells,which leads to angiogenesis.Besides to the vascular endothelial growth factor,the effect of adenosine on angiogenesis is increasingly concerned.Objective This study was to invastigate the relationship of biological behaviour of human microvascular endothelial cells (HMEC-1) under the hypoxia and adenosine,and to explore the effect of adenosine on angiogenesis under hypoxia.Methods HMEC-1 cell line was cultured in vitro,and the cells were divided into normoxia group and hypoxia group.The cells in the normoxia group were cultured under the 5％ CO2 environment,and those in the hypoxia group were cultured under the 1％ O2,94％ N2 and 5％ CO2 environment.The proliferation ability and percentage of the cells were assayed by cell counting kit-8 (CCK8) and EDU.The migration number and invasive number of the cells were detected by transwell chamber.The expressions of CD39 and CD73 proteins in the cells were tested by Western blot and immunofluorescence technique,and the level of adenosine was measured by high performance liquid chromatography (HPLC).Results The proliferation values (absobancy) were 0.715-±0.067 and 0.821 ±0.056 in the normoxia group and the hypoxia group in 12 hours after culture,and those in 24 hours were 0.946±0.028 and 0.998±0.028,showing significant increase in the hypoxia group compared with the normoxia group (t12h =3.805,t24h =3.222,all at P ＜ 0.01).The precentage of the proliferation in the hypoxia group was evidently higher than that in the normoxia group (t =-6.868,P＜0.01).The number of the cell migration and invasion in 24 hours after culture was 185.3 ± 10.594 and 74.2± 10.741 respectively in the normoxia group,and that in the hypoxia group was 300.7±22.853 and 107.5±7.007,with significant differences between the two groups (t=-12.124,-6.367,both at P＜0.01).The expression levels of CD39 and CD73 proteins in the hypoxia group were significantly higher than those in the normoxia group in bothl2 hours and 24 hours after culture(all at P＜0.05),and the adenosine content in the cells is gradually increased in 2,6,12,24 and 36 hours after culture,with the highest content in 36 hours.The adenosine content was significantly higher in the hypoxia group than in the normoxia group at various time points (t2h =2.469,P =0.017;t6h =5.442,P＜0.001;t12h =3.841,P＜0.001;t24h =4.458,P＜0.001;t36h =2.757,P =0.008;t48h =3.319,P =0.002).Conclusions Compared with the normoxia group,the proliferation,migration and invasion abilities of HMEC-1 are stronger,meanwhile,the expression of CD39 and CD73 as well as the adenosine level in the cells are all increased under the hypoxic condition.It is suggested that the activation of human microvascular endothelial cells might be significantly related to the level of adenosine and its key enzymes.

Background Retinal ganglion cell (RGCs) death following ischaemic insult is the major cause of a number of vision-threatening diseases.Recent studies confirmed that micro RNA (miR-30b) can alleviate hypoxy-induced cardiac injury.However,whether miR-30b can protect RGCs against oxygen-glucose deprivation damage is still not ellucidated.Objective The aim of this study was to investigate the protective effect of miR-30b on RGCs damage caused by oxygen-glucose deprivation.Methods The retinas were isolated from the eyeballs of eight SD rats aged postnatal 24 hours and RGCs were primarily cultured.The cells were divided into the recombinant adeno-associated virus (rAVV) control group,rAAV-miR-30b mimic group and AAV-miR-30b inhibitor group.Then the cells were transfected using rAVV-miR plasmid,rAAV-miR-30b mimic plasmid and AAV-miR-30b inhibitor plasmid,respectively for 6 days with the RGCs ∶ AAV as 1 ∶ 10 000.The cells were cultured with low glucose medium in hypoxygen incubator (5％ CO2,17％ N2,3％ O2) or 5％ CO2 incubator respectively for 24 hours.Cell viability was detected by cell counting kit-8 assay.The expression of Tubulin Ⅲ,a neuron specific marker,was detected by immunofluorescence technology to evaluate the survival of RGCs.The apoptosis and necrosis of the cells were assessed by Hoechst/PI double staining.Results The RGCs grew well with round shape and 1 3 processes 7 days after cultured in the normal cells.However,the RGCs were diminished and the cell process disrupted in the oxygen-glucose deprivation group.The relative vability of the cells was 3.310-±0.162 in the rAAV-miR-30b mimic group,which was significantly higher than 0.949±0.141 in the rAAV-miR-30b inhibitor group and 0.900±0.181 in the rAAV-miR control group(t=10.508,10.296,both at P＜0.001).It was positively expressed in survival RGCs,with the red fluorescence.The number of Tubulin Ⅲ+ cells was (13.800± 1.924)/field in the rAAV-miR-30b mimic group,showing a significant increase in comparison with (0.600±0.548)/field in the rAAV-miR-30b inhibitor group and (0.800± 1.304)/field in the rAAV-miR control group (t =15.141,14.912,both at P ＜ 0.001).Significant differences were found in the apoptosis rate and necrosis rate among the rAAV-miR-30b mimic group,rAAV-miR control group and PBS group (F=10.851,P=0.002;F=6.378,P=0.013),and the apoptosis rate and necrosis rate in the rAAV-miR-30b mimic group were considerably lower than those in the rAAV-miR control group and PBS group (all at P＜0.05).Conclusions The oxygen-glucose deprivation models can be established in RGCs by hypooxygic and low-glucose cultivation.rAAV encoding miR-30b mimics transfection can protect RGCs against oxygen-glucose deprivation damage.

Objective In order to reduce the toxicity and still maintain the bioactivity of the medicinal plant Tripterygium wilfordii(TW),the whole plant biotransfomed method was employed.By observing the pharmacological activities and the toxicity of MeOH extracts from the root of T.wilfordii that were biotransformed by the fungi Aspergillums spp.and Rhizopus spp.,the best biotransformed fungi strain has been selected based on the experimental data.Methods Applying the models of acute and chronic inflammation, the weight of immune organ and cell immunity were used to study the pharmacological activities,and ip injection to observe the acute toxicity.Results The anti-inflammation and immunosuppression have been increased slightly and the toxicity decreased significantly after being biotransformed by Aspergillus spp.(TW1),while the anti-inflammation,immunosuppression,and the toxicity have been declined after being biotransformed by Rhizopus spp.(TW2).Conclusion Although both selected fungi could change the pharmacological activities and toxicity of the T.wilfordii,Aspergillus spp.is better than Rhizopus spp.

Objective:To explore the application of mixed BOPPPS (bridge-in, objective, pre-assessment, participatory-learning, post-assessment, summary) combined with case analysis teaching in nursing practice teaching in the department of endocrinology.Methods:Forty-five nurses of Batch 2017 who practiced in the Department of Endocrinology were selected as the control group, using traditional teaching; 45 practical nursing students of Batch 2018 were selected as the research group, using mixed BOPPPS combined with case analysis teaching. After the teaching, the teaching effect was evaluated by the theoretical and practical skill operation assessments, and the satisfaction with the teaching was investigated through the questionnaire. SPSS 22.0 was used for t test and chi-square test. Results:The scores of basic theory [(28.31±2.07) vs. (24.66±2.15)] and professional theory [(67.01±3.05) vs. (55.62±3.20)] in the theoretical assessment of practical nurses in the research group were higher than those in the control group, with significant differences ( P<0.05); the scores of each sub-item of the practical skills operation assessment were better than those in the control group, with significant differences ( P<0.05); in terms of teaching satisfaction, the research group was better than the control group in such 10 aspects as whether to improve the level of theoretical and practical skills operation, whether to improve the awareness of humanistic care and whether to improve the nurse-patient communication ability, etc., with significant differences ( P<0.05). Conclusion:The BOPPPS combined with case analysis teaching significantly improves the theoretical and practical skills and comprehensive literacy of the endocrinology practical nursing students, and the teaching satisfaction is significantly better than that of the traditional teaching, which is of promotion value.

Objective:To study the duration of invasive mechanical ventilation (MV) and its influencing factors after neonatal abdominal surgery under general anesthesia in neonatal intensive care unit (NICU).Methods:From January 2018 to December 2020, neonates received abdominal surgery under general anesthesia and needed endotracheal intubation and MV after surgery in NICU of our hospital were retrospectively studied. According to MV duration, the neonates were assigned into <72 h group and ≥72 h group. Multivariate logistic regression was used to analyze the risk factors of postoperative MV duration.Results:A total of 113 neonates were enrolled, including 57 male (50.4%) and 56 female (49.6%). The gestational age was (35.7±3.6) weeks, the birth weight was (2 497±933) g, the average operation age was 9.9(3.6, 22.2) d and the average hospital stay was 22.0(12.0,37.0) d. Congenital intestinal obstruction (37/113, 32.7%) was the most common diagnoses on discharge, followed by neonatal necrotizing enterocolitis(28/113,24.8%) and gastrointestinal perforation (18/113,15.0%). The duration of operation was 80.0 (55.8,117.3) min. All neonates needed MV with endotracheal intubation. The duration of postoperative respiratory support was 30.0(7.0,84.5) h. 48 neonates (42.5%) had endotracheal intubation removed within 24 h after surgery. Multivariate logistic regression analysis showed that preoperative respiratory support ( P=0.004), congenital heart disease( P=0.013) and intravenous midazolam ( P=0.032) were independent risk factors for prolonged postoperative MV. Conclusions:The need of preoperative respiratory support, congential heart disease and intravenous midazolam were independent risk factors for the duration of postoperative MV after neonatal abdominal surgery under general anesthesia.

Objective: To explore the gene lifestyle interaction of ATP2B1-eNOS pathway gene polymorphisms on blood pressure. Methods: Using the convenient cluster sampling method, a total of 872 junior middle school students from 3 school in July to August 2019, were included in the final analysis. The survey included questionnaire investigation, anthropometry measurement and blood sample collection. After DNA was extracted from peripheral blood samples, the gene polymorphisms ( ATP2B1 gene rs 17249754 and rs 2070759, eNOS gene rs 1799983 and rs 2070744) were genotyped. Logistic regression model was used to analyze the association between gene polymorphism and blood pressure phenotypes. Results: The prevalence of high blood pressure was 9.52 % in adolescents(9.15% in boys and 9.87% in girls),with no significant sex difference ( χ 2=0.13, P =0.72). There were statistically significant differences between boys and girls in age, systolic blood pressure (SBP), diastolic blood pressure (DBP), body mass index (BMI) classification, birth weight, daily school physical exercise time and daily playing video games time ( P <0.05). Logistic regression analysis showed that, eNOS gene rs 2070744 polymorphism was associated with high blood pressure (HBP) under the recessive model, and the risk of HBP in CC genotype carriers were higher than that TT/TC genotype carriers ( OR=3.88, 95%CI =1.00-15.02, P < 0.05 ). The results of gene lifestyle interaction showed that ATP2B1 gene rs 2070759 polymorphism gene had an interaction with the time of physical exercise in school ( P interaction =0.05). In the subgroup with daily physical exercise time at sch ool ＜1 hour , the TT/TG genotype carriers were associated with increased risk of HBP compared with GG genotype carriers( OR= 2.65 , 95%CI =1.11-6.30, P <0.05). But in the subgroup with daily physical exercise time in school ≥1 hour, rs 2070759 was not significantly associated with HBP. Conclusion eNOS/rs 2070744 polymorphisms are associated with risk of HBP among adolescents. There is significant interaction between ATP2B1 gene rs 2070759 polymorphism and physical exercise time in school on HBP. Adolescents should spend more time on physical activity in school, which will help to maintain normal blood pressure level.

Objective:To evaluate Zhuang medicine Fuzheng compound combined with epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in the treatment of advanced epidermal growth factor receptor (EGFR) sensitive mutant non-small cell lung cancer (NSCLC).Methods:A total of 120 patients with advanced NSCLC who met the inclusion criteria from June 2019 to May 2020 in Guangxi International Zhuang Medical Hospital were divided into 2 groups according to the random number table method, with 60 in each group. The control group was treated with TKIs, and the observation group was treated with Zhuang medicine Fuzheng compound combined with EGFR-TKIs. TCM syndrome scores were compared, and the quality of life of the patients was assessed by the Quality of Life Scale (QLQ-C30). The serum levels of carcinoembryonic antigen (CEA), squamous cell carcinoma associated antigen (SCC-Ag) and carbohydrate antigen 50 (CA50) were detected by radioimmunoassay, and the levels of CD3 +, CD4 +, and CD8 + were detected by flow cytometry, and the CD4 +/CD8 + ratio was calculated. The adverse reactions during the treatment were observed and recorded. Results:The objective remission rate in the observation group was 66.7% (40/60) and the disease control rate was 81.7% (49/60), while in the control group were 48.3% (29/60) and 63.3% (38/60), respectively.The differences were statistically significant ( χ2 values were 4.13 and 5.06, P values were 0.042 and 0.025, respectively). After treatment, the scores of chest tightness, shortness of breath, blood in sputum, mental fatigue in the observation group were significantly lower than those in the control group ( t values were 8.72, 5.02, 5.47, all Ps<0.001), After treatment, QLQ-C30 score in the observation group was significantly higher than that of the control group ( t=5.21, P<0.01). After treatment, CEA [(31.45±4.56) mU/L vs. (38.98±5.71) mU/L, t=7.98], SCC-Ag [(4.87±0.93) μg/L vs. (7.29±1.25) μg/L, t=12.03], CA50 [(58.27±7.14) U/L vs. (66.48±7.94) U/L, t=5.96] levels were significantly lower than those in the control group ( P<0.01); CD3 +[(52.43±5.01)% vs. (48.56±4.87)%, t=4.29], CD4 + [(54.89±5.03)% vs. (51.09±5.22)%, t=4.06], CD4 +/CD8 + [(1.95±0.28) vs. (1.65±0.27), t=5.97] significantly higher than those in the control group ( P<0.01), CD8 + [(28.12±2.70)% vs. (31.23±2.64)%, t=6.38] significantly lower than that of the control group ( P<0.01). During the treatment period, the incidence of adverse reactions in the observation group was 13.3% (8/60) and that in the control group was 8.3% (5/60), with a statistically significant difference between two groups ( χ 2=0.78, P=0.378). Conclusion:The Zhuang medicine Fuzheng compound combined with EGFR-TKIs can reduce the level of tumor markers in patients with advanced EGFR-sensitive mutant NSCLC, improve patients' TCM syndromes, quality of life, enhance patient immunity, and improve efficacy.

The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.

[Abstract] Objective: To investigate the miRNAs that can intervene Mcl-1 expression in HBV-related liver cancers and to study their synergistic anti-cancer effect with sorafenib. Methods: The expressions of miR-29, miR-101 and miR-193b in HepG2.2.15 (HBV positive) and HepG2.vc (HBV negative) cells were detected by qPCR. miRNA mimics of low expressed genes in HepG2.2.15 cells were synthesized and transfected into HepG2.2.15 and HepG2.vc cells, respectively. qPCR was used to detect target miRNA expression. Western blotting was used to detect the expression of mcl-1 protein in cells before and after transfection.At the same time, (1×10-9)~(1× 10-3) mol/L of sorafenib was add to both transfected and non-transfected HepG2.2.15 and HepG2.vc cells; 72 h later, the IC50 and cell apoptosis was evaluated. Results: The expression of miR-193b in HepG2.2.15 cells was significantly lower than that in HepG2.2.15 cells (P <0.05). The expression of miR-193b in HepG2.2.15 cells and HepG2.2.15 cells was significantly higher after miR-193b mimics transfection (P <0.05). Compared with HepG2.vc cells, the expression of Mcl-1 protein in HepG2.2.15 cells was significantly increased (P <0.05). The expression of Mcl-1 protein in HepG2.2.15 and HepG2.vc cells was significantly decreased after miR-193b mimics transfection (P<0.05). After miR-193b mimics transfection, sorafenib could significantly increase apoptosis rate of both HepG2.2.15 and HepG2.vc cells. Conclusion: The low susceptibility of HBV-related liver cancer to sorafenib may be related with the low expression of miR-193b in cancer cells. Mcl-1 might be used as a target of miR-193b, and miR-193b mimics have a significant synergistic effect with sorafenib.

Objective: To investigate the clinical value of combined detection of serum miR-378 and miR-21 in gastric cancer (GC). Methods: Eighty-seven patients with GC and 78 patients with colorectal cancer(CRC) from National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences were selected, 83 individuals undergoing healthy physical examination were selected as the healthy controls. The levels of serum miR-378 and miR-21 were detected by quantitative real-time PCR (RT-qPCR) (result data were transformed as log2 for analysis). Results: Relative expression levels of miR-378 in the serum were -1.24, -3.25 and -2.73 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-378 were significantly decreased in GC and CRC patients (both P<0.05). Relative expression levels of miR-21 in the serum were 0.11, 2.34 and 2.47 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-21 were significantly up-regulated in GC and CRC patients (both P<0.05). Moreover, the serum level of miR-378 in GC patients was inversely associated with tumor clinical stage (P<0.05). However, the level of miR-21 showed no significant differences among patients with different clinical and pathological characteristics (all P>0.05). The area under the receiver operating characteristic curve (AUC), sensitivity and specificity of miRNA-378 to diagnose GC was 0.770, 82.0% and 66.0%, respectively, and were 0.900, 85.0%, and 88.0% of miR-21, respectively. The AUC, sensitivity and specificity of combined detection of serum miR-378 and miR-21 to diagnose GC were 0.930, 92.0% and 87.0%, respectively, while the AUC of combined detection of serum CEA and CA-199 was 0.767, the AUC of combined all of the four factors was 0.946. Conclusion The combined detection of serum miR-378 and miR-21 have a certain effect on diagnosis of GC.