AIM: To construct a knock-out vector for ?~(maj) & ?~(min) gene of ?-globin gene and establish embryonic stem cell lines by gene knock-out in ES level for generating gene knock-out animal model. METHODS: Two homologous sequences 1.7 kb and 7.0 kb were inserted into basic plasmic vector pNTK, which contains positive and negative selection system of the neomycin resistance (Neo) and herpes simplex virus thymidine kinase (TK) resistant gene to construct a gene knock-out recombinant plasmic vector nominated ?-pNTK, which was certified by polymerase chain reaction, enzyme digested and DNA sequencing. The ?-pNTK that contains two homologous sequence was linearized with Sal I and introduced into the D3 line of ES cells by electroporation. Positive cloning of ES cells was selected with positive and negative selection system of G418 600 mg/L and 2.5 mol/L Gancyclovir for four weeks. ES cells clone were picked, explanded and certified by polymerase chain reaction, Southern blotting. Examination of the undifferentiated state and pluripotential characteristics of the cell lines were made with the alkaline phosphatase (AKP) staining and embryoid bodies formation test and teratomas formation in nude mice. RESULTS: A knock-out recombinant plasmic vector for ? maj & ? min gene of ?-globin gene, ?-pNTK, was constructed in C129 mouse, which contains 8.7 kb length inserted homologous sequence and deleted most part of the ?~(maj) & ?~(min) gene. 8 strains of positive ES cells cloning were obtained, all of them were testified positive with polymerase chain reaction and Southern blotting. The results of AKP staining, embryoid bodies formation in vitro and teratomas formation in nude mice showed undifferentiated state and pluripotential characteristic of cell line. CONCLUSIONS: A knock-out recombinant plasmic vector of ?~(maj) & ?~(min) of ?-globin gene in C129 mouse was constructed, in which a knock-out ?~(maj) & ?~(min) gene of ?-globin gene ES-D3 cell line was established by electroporation, characterization of these cells show a prospect utility in producing gene knock-out mice in blastocysts stages.

PURPOSE: C-terminal binding protein 1 (CtBP1) is a transcriptional co-repressor that is overexpressed in many cancers. CtBP1 transcriptionally represses a broad array of tumor suppressors, which promotes cancer cell proliferation, migration, invasion, and resistance to apoptosis. Recent studies have demonstrated that CtBP1 is a potential target for cancer therapy. This study was designed to screen for compounds that potentially target CtBP1. METHODS: Using a structure-based virtual screening for CtBP1 inhibitors, we found protocatechuic aldehyde (PA), a natural compound found in the root of a traditional Chinese herb, Salvia miltiorrhiza, that directly binds to CtBP1. Microscale thermophoresis assay was performed to determine whether PA and CtBP1 directly bind to each other. Further, clustered regularly interspaced short palindromic repeats associated Cas9 nuclease-mediated CtBP1 knockout in breast cancer cells was used to validate the CtBP1 targeting specificity of PA. RESULTS: Functional studies showed that PA repressed the proliferation and migration of breast cancer cells. Furthermore, PA elevated the expression of the downstream targets of CtBP1, p21 and E-cadherin, and decreased CtBP1 binding affinity for the promoter regions of p21 and E-cadherin in breast cancer cells. However, PA did not affect the expression of p21 and E-cadherin in the CtBP1 knockout breast cancer cells. In addition, the CtBP1 knockout breast cancer cells showed resistance to PA-induced repression of proliferation and migration. CONCLUSION Our findings demonstrated that PA directly bound to CtBP1 and inhibited the growth and migration of breast cancer cells through CtBP1 inhibition. Structural modifications of PA are further required to enhance its binding affinity and selectivity for CtBP1.

【Objective】 To strengthen the management of transfusion adverse events, so as to reduce the occurrence of medical damage and accidents, and guarantee the safety of blood transfusion. 【Methods】 The adverse events of blood transfusion reported in our hospital from July 2016 to December 2022 were collected, the reasons were tracked, and continuous improvements were made. 【Results】 From 2016 to 2022, a total of 315 transfusion adverse events were reported, including 233(73.97%, 233/315) cases of transfusion reactions and 82(26.03%, 82/315) transfusion adverse events. There were 271 328 transfusion cases in the same period, and the incidence of transfusion reactions was 0.858 7‰(233/271 328). The number of transfusion application was 129 887, and the incidence of transfusion adverse event is 0.631 3‰(82/129 887). Sixty-eigtht(82.93%, 68/82) cases of transfusion adverse events were caused by human factors, while the other 14(17.07%, 14/82) cases were non-human factors. According to the linear regression analysis, we have concluded that the year is a significant indicator for transfusion reaction rates (P<0.05), but not for transfusion adverse event rates (P>0.05). 【Conclusion】 Strengthening the management of reporting adverse events in clinical blood transfusion, monitoring the incidence, analyzing and improving different types of adverse events by management tools can reduce the medical risks of blood transfusion and help to guarantee the safety of clinical blood transfusion.

【Objective】 To investigate the factors influencing the length of hospital stays of the acute fatty liver of pregnancy (AFLP) patients, so as to establish the prediction model. 【Methods】 A total of 49 patients diagnosed as AFLP)in ShenZhen People’s Hospital between January 2008 and January 2023 were retrospectively analyzed. According to the median length of hospital stays, the patients were divided into two groups: Group A(n=21)and Group B(n=28). Preoperative general laboratory data, clinical features and postpartum adverse outcomes in both groups were analyzed. Multivariate binary logistic regression was used to analyze the independent factors affecting the length of hospital stays for AFLP, and a prediction model for hospitalization time was established. 【Results】 Comparison between Group B and Group A were as follows: hospital stays(d)(15.5 vs 8), preoperative icterus(%)[16(57.1%)vs 3(14.3%)], thrombin time(TT)(s)(24.2 vs 21.3), prothrombin time(PT)(s)(16.8 vs 15.3), activated partial thromboplastin time(APTT)(s)(52.3 vs 40.7), total bilirubin(TBIL)(μmol/L)(77.2 vs 45.2), indirect bilirubin(IBIL)(μmol/L)(21.2 vs 10), creatinine(Cre)(μmol/L)[(171.97±53.34) vs (131.81±45.06]), TT extension(%)[24(85.7%)vs 11(52.4%)], APTT extension(%)[27(96.4%)vs 7(33.3%)], IBIL elevation(%)[19(67.9%)vs 4(19%)], Cre concentration rise(%)[21(75%)vs 8(38%)], number of postpartum plasma exchange sessions(%)[23(82.1%)vs 5(23.8%)], postpregnancy co-infection phenomenon(%)[21(75%)vs 4(19%)], with Group B significantly higher than Group A. The preoperative platelet count(×10⁹/L)(128 vs 221)and the concentration of fibrinogen(g/L)[0.9 vs 1.6] in Group B were significantly lower than those in Group A. Univariate logistic regression analysis showed that preoperative icterus, postpregnancy co-infection phenomenon, number of postpartum plasma exchange sessions, preoperative TT extension, preoperative APTT extension, Cre concentration rise were influencing factors for the hospital stays of AFLP patients. According to the minimum result of Akaike information criterion, the multivariate binary logistic regression analysis (step-wise selection) showed that the number of postpartum plasma exchange sessions, icterus, preoperative APTT extension were the independent risk factor influencing the hospital stays of AFLP patients, and the logistic regression prediction model was established by incorporating the above three factors. Regularization techniques were further employed in linear regression to address and assess overfitting issues. Additionally, the confidence interval for the estimated effect sizes in each model have been acquired by bootstrapping techniques. 【Conclusion】 Preoperative icterus, preoperative APTT extension(APTT>43s)and the number of postpartum plasma exchange sessions were the independent risk factor influencing the hospital stays of AFLP patients and the logistic regression prediction model with high predictive effectiveness was established successfully.

Objective:To investigate the prevalence of hyperuricemia (HUA) in Bozidun Kirghiz township of Xinjiang Aksu region, and to explore the risk factors for the occurrence of HUA in the local area.Methods:A cross-sectional survey study was conducted by randomly selecting 9 villages in Bozidun Kirgiz Township by the whole-group sampling method and questionnaire were distributed to the households. The questionnaire included: demographic information, history of past illness, personal history, and all subjects were measured for height, weight, blood pressure, abdominal circumference, etc. The diagnostic of HUA if the serum uric acid (SUA) level >420 μmol/L in men or >360 μmol/L in women. The incidences of HUA in different age, sex, food type and life style behavior were analyzed. T test, non-parametric test and Chi-square test were used to analyze the differences among the groups, and logistic regression was used to analyze the risk factors. Results:①A total of 2 138 subjects were surveyed, among which 68 patients were with HUA, the prevalence of HUA in Bozidun Kirghiz township, Aksu region in the general population was 3.18%(68/2 138); the prevalence rate in men was 4.60%(45/978), 45 patients were identified; and the prevalence rate in women was 1.98%(23/1 160), 23 patients were identified. The peak age of HUA in male and female patients was 51~60 years old. ②The prevalence of HUA was lower in those who consumed dairy products ( χ2=6.91, P=0.017), nuts ( χ2=8.43, P=0.038) and eggs ( χ2=7.38, P=0.023), and lower in those who consumed more. Different intake of cereals ( χ2=0.87, P=0.647), meat( χ2=0.82, P=0.662), vegetables and fruits( χ2=5.22, P=0.073) had no effect on the prevalence of HUA.③In terms of different life behaviors, the prevalence of HUA in men who had been smoking was higher than those who had never smoked (57.78%, 28.89%, 13.33%, χ2=8.16, P=0.017). In the relationship between drinking and HUA, the prevalence rates of male always drinking, ever drinking and never drinking were 80.00%, 11.11% and 3.89%, respectively, the difference was statistically significant ( χ2=6.67, P=0.038). ④Multi-factor logistic regression analysis showed that high BMI, old age, high TG, increased Cr and increased WBC were risk factors for the occurrence of HUA [ OR(95% CI)=1.13(1.04, 1.23), 1.03(1.00,1.05),1.39(1.00, 1.93), 1.03(1.02, 1.05), 1.27(1.07, 1.49), all P<0.05]. Conclusion:The prevalence of HUA in Bozidun Kirgiz township in Aksu prefecture of Xinjiang is lower than that in other areas with continental climate. High BMI, old age, high TG, increased Cr and increased WBC count are risk factors for the development of HUA .

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult ; Asian Continental Ancestry Group* ; China ; Comorbidity ; Developed Countries ; Developing Countries ; Diagnosis* ; Epidemiologic Studies ; Epidemiology ; Global Health ; Humans ; Hypersensitivity* ; Prevalence ; Rhinitis, Allergic*

Objective:To assess the effectiveness of a 6-month Ba Duan Jin exercise program in improving the balance of community-dwelling older adults.Methods:A two arms, parallel-group, cluster randomized controlled trial was conducted in 1 028 community residents aged 60-80 years in 40 communities in 5 provinces of China. Participants in the intervention group (20 communities, 523 people) received Ba Duan Jin exercise 5 days/week, 1 hour/day for 6 months, and three times of falls prevention health education, and the control group (20 communities, 505 people) received falls prevention health education same as the intervention group. The Berg balance scale (BBS) score was the leading outcome indicator, and the secondary outcome indicators included the length of time of standing on one foot (with eyes open and closed), standing in a tandem stance (with eyes open and closed), the closed circle test, and the timed up to test.Results:A total of 1 028 participants were included in the final analysis, including 731 women (71.11%) and 297 men (28.89%), and the age was (69.87±5.67) years. After the 3-month intervention, compared with the baseline data, the BBS score of the intervention group was significantly higher than the control group by 3.05 (95% CI: 2.23-3.88) points ( P<0.001). After the 6-month intervention, compared with the baseline data, the BBS score of the intervention group was significantly higher than the control group by 4.70 (95% CI: 4.03-5.37) points ( P<0.001). Ba Duan Jin showed significant improvement ( P<0.05) in all secondary outcomes after 6 months of exercise in the intervention group compared with the control group. Conclusions:This study showed that Ba Duan Jin exercise can improve balance in community-dwelling older adults aged 60-80. The longer the exercise time, the better the improvement.

ObjectiveTo explore the effects of phillygenin （PHI） on the inflammation in L02 cells induced by lipopolysaccharide （LPS） and adenosine triphosphate （ATP） and the expression of purinergic 2X7 receptor （P2X7R）， NOD-like receptor family pyrin domain containing 3 （NLRP3）， and nuclear factor kappa B （NF-κB） expression. MethodIn this study， the inflammation model was induced in L02 cells by 100 μg·L^-1 LPS treatment for 24 h and 5 mmoL·L^-1 ATP treatment for 5 h. The cells in the PHI groups were cultured with PHI （100， 50， 25 mg·L^-1） for 6 h in the LPS treatment period， followed by LPS treatment for another 18 h. After ATP treatment for 5 h， the mRNA and protein expression of interleukin-1β （IL-1β）， interleukin-18 （IL-18）， P2X7R， NLRP3， Caspase-1 precursor （pro-Caspase-1）， cleaved Caspase-1， NF-κB， and NF-κB inhibitor protein α （IκBα） in L02 cells was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. Molecular docking was used to predict whether P2X7R could bind to PHI， and DCFH-DA was employed to detect the accumulation of reactive oxygen species （ROS） in cells. P2X7R was silenced by small interfering ribonucleic acid （siRNA）， and then the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was detected by Real-time PCR. ResultReal-time PCR and Western blot showed that compared with the normal group， the model group showed increased expression of IL-1β and IL-18 （P<0.05）， and compared with the model group， the PHI groups showed down-regulated IL-1β， IL-18 mRNA and protein expression （P<0.05）. Molecular docking suggested a good binding effect of PHI to P2X7R. Real-time PCR and Western blot analysis showed that the expression of P2X7R in the model group was significantly up-regulated compared with that in the normal group （P<0.05）， and compared with the model group， the PHI groups showed down-regulated mRNA and protein expression of P2X7R （P<0.05）. DCFH-DA results showed that compared with the normal group， the model group showed increased content of ROS （P<0.05）， and compared with the model group， the PHI groups decreased the accumulation of ROS （P<0.05）. As demonstrated by Real-time PCR and Western blot， compared with the normal group， the model group showed increased expression of NLRP3 inflammasome and NF-κB （P<0.05）， and compared with the model group， the PHI groups significantly decreased the mRNA and protein expression of NLRP3 and cleaved Caspase-1， and up-regulated the mRNA and protein expression of NF-κB and IκBα （P<0.05）. Real-time PCR analysis showed that compared with the results in the model group， after silencing P2X7R by siRNA， the mRNA expression of IL-1β， IL-18， P2X7R， NLRP3， Caspase-1， NF-κB， and IκBα was decreased （P<0.05）. PHI exerted the same effect， and the mRNA expression was further reduced after the combination of them. ConclusionPHI is presumed to suppress the expression of the NLRP3/NF-κB signaling pathway by down-regulating upstream P2X7R to alleviate the LPS/ATP-induced inflammation in L02 cells， suggesting that P2X7R may be the target of PHI against inflammation.