ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

Objective To explore the changes of hemodynamics in pregnant women with uncomplicated dichorionic(DC)twin pregnancy and monochorionic(MC)twin pregnancy,so as to better perform prenatal monitoring.Methods A study was conducted to collect 64 pregnant women with uncomplicated twin pregnancy(41 cases of DC and 23 cases of MC)in the Obstetrics and Gynecology Hospital,Fudan University from May 2020 to Sept 2021,and 144 pregnant women with uncomplicated singleton pregnancies in the same period were selected as the control group.During the second trimester(20-28 weeks),conventional echocardiography was performed in pregnant women,and singleton pregnancy was used as the control.The left ventricular systolic,left ventricular diastolic,hemodynamic and cardiac structural parameters of DC and MC twin pregnancy were studied.Results At 20-28 weeks of gestation,compared with singleton pregnancies,the heart rate,mean arterial pressure,cardiac output,cardiac index,stroke volume,left ventricular mass,stroke work index parameters of twin pregnancy were significantly increased,and the total vascular resistance was significantly decreased,and the differences were statistically significant(P<0.05).The distribution of maternal hemodynamic parameters was similar in pregnant women with DC twin and MC twin pregnancy.Compared with DC twin pregnancy,MC twin pregnancy showed a significant increase in cardiac output(5.76 L/min vs.5.36 L/min,P=0.031).Total vascular resistance significantly decreased(1 270 vs.1 407,P=0.037).Conclusion Compared with singleton pregnancy,the hemodynamics of twin pregnancy significantly changed,which ensured the growth and development of the fetus by providing sufficient uteroplacental circulation.The cardiovascular adaptation patterns of DC and MC twin pregnancy were similar,but MC twin pregnancy had higher cardiac output and lower total vascular resistance.Monitoring the cardiac function of twin pregnancy,especially MC twin pregnancy,is very important for the safety of pregnant women.

To date, there is little information about the demography of vasectomy reversal (VR) patients or the factors currently influencing VR effectiveness in China, especially after the universal two-child policy was released in 2015. In this research, demographic data and perioperative medical records of VR patients were extracted from seven major hospitals in different provinces or municipalities of China. Meanwhile, a telephone survey of the patients was conducted to collect follow-up information. Eventually, 448 VR cases from the past 13 years were included. The results were analyzed by stratified comparison to investigate factors that can influence postoperative vas deferens patency and pregnancy rate. Appropriately statistical methods were used, and all of the protocols were approved by the Ethics Committees of the institutes in this research. The results showed that the annual operation volume of VR quadrupled after the two-child policy was implemented. Nonmicrosurgery and a long duration of vasectomy were significantly associated with a lower patency rate. A follow-up survey showed that the general postoperative pregnancy rate was 27.2%. For female partners over the age of 35 years, the postoperative pregnancy rate showed a more severe decline, but only 35.5% of them had been given a fertility examination before their husbands' VR surgery. Our work revealed that more patients in China have been demanding VR in recent years. High-quality microsurgery and a short duration of vasectomy are crucial for restoring patency by VR. Clinical andrologists should perform a preoperative fertility evaluation of the patients' female partners.
Male ; Pregnancy ; Humans ; Female ; Adult ; Vasovasostomy ; Retrospective Studies ; Vas Deferens/surgery* ; Vasectomy ; China/epidemiology*

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.

OBJECTIVE: To evaluate the effects of auricular acupoint bloodletting (AB) and auricular acupressure (AA) on sleep quality and the levels of melatonin (MT), glutamic acid (Glu), and γ -aminobutyric acid (GABA) in college students with primary insomnia, and to explore the possible mechanism. METHODS: Totally 74 college students at Hebei University of Chinese Medicine with primary insomnia were selected from October 2019 to October 2020. All patients were assigned to AB+AA group (37 cases, received combination of AB and AA) and AA group (37 cases, received only AA on the same acupoints) by a random number table. Each group was treated twice a week for 4 weeks. The Pittsburgh Sleep Quality Index (PSQI) score, Chinese medicine (CM) syndrome score, total effective rate, serum concentrations of MT, Glu, and GABA, and Glu/GABA ratio were compared between the two groups after treatment and at follow-up. The safety of therapy was also evaluated. RESULTS: After 4-week treatment, the total scores of PSQI, each PSQI component score, and the CM syndrome scores in both groups all decreased (P<0.05); the serum MT concentrations in both groups all increased (P<0.05). The concentrations of Glu and GABA in the AB+AA group were significantly higher than those in the AA group after treatment (P<0.05). However, there was no significant difference in the ratio of Glu/GABA in both groups before and after treatment (P>0.05). At follow-up, the CM syndrome score in the AB+AA group was significantly lower than that in the AA group (P<0.05). There was no significant difference between the two groups in total effective rates and adverse events (P>0.05). CONCLUSIONS Both AB+AA and AA can relieve insomnia symptoms, but a stronger long-term effect were observed for AB+AA. AB+AA can promote the secretion of MT, increase the levels of Glu and GABA more effective than AA, and regulate their imbalance, and thus it may be benificial for treating insomnia.
Humans ; Acupressure ; Acupuncture Points ; Bloodletting ; Sleep Initiation and Maintenance Disorders/therapy* ; Sleep Quality ; Syndrome ; Students ; gamma-Aminobutyric Acid

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child ; China/epidemiology* ; Cryptorchidism/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genital Diseases, Male ; Genotype ; Humans ; Hypospadias/genetics* ; Male ; Membrane Proteins/genetics* ; Penis/abnormalities* ; Phenotype ; Retrospective Studies ; Steroid 21-Hydroxylase/genetics*

BACKGROUND: Bendamustine was approved in China on May 26th, 2019 by the National Medical Product Administration for the treatment of indolent B-cell non-Hodgkin lymphoma (NHL). The current study was the registration trial and the first reported evaluation of the efficacy, safety, and pharmacokinetics of bendamustine in Chinese adult patients with indolent B-cell NHL following relapse after chemotherapy and rituximab treatment. METHODS: This was a prospective, multicenter, open-label, single-arm, phase 3 study (NCT01596621; C18083/3076) with a 2-year follow-up period. Eligible patients received bendamustine hydrochloride 120 mg/m2 infused intravenously on days 1 and 2 of each 21-day treatment cycle for at least six planned cycles (and up to eight cycles). The primary endpoint was the overall response rate (ORR); and secondary endpoints were duration of response (DoR), progression-free survival (PFS), safety, and pharmacokinetics. Patients were classified according to their best overall response after initiation of therapy. Proportions of patients in each response category (complete response [CR], partial response [PR], stable disease, or progressive disease) were summarized along with a two-sided binomial exact 95% confidence intervals (CIs) for the ORR. RESULTS: A total of 102 patients were enrolled from 20 centers between August 6th, 2012, and June 18th, 2015. At the time of the primary analysis, the ORR was 73% (95% CI: 63%-81%) per Independent Review Committee (IRC) including 19% CR and 54% PR. With the follow-up period, the median DoR was 16.2 months by IRC and 13.4 months by investigator assessment; the median PFS was 18.6 months and 15.3 months, respectively. The most common non-hematologic adverse events (AEs) were gastrointestinal toxicity, pyrexia, and rash. Grade 3/4 neutropenia was reported in 76% of patients. Serious AEs were reported in 29 patients and five patients died during the study. Pharmacokinetic analysis indicated that the characteristics of bendamustine and its metabolites M3 and M4 were generally consistent with those reported for other ethnicities. CONCLUSION: Bendamustine is an active and effective therapy in Chinese patients with relapsed, indolent B-cell NHL, with a comparable risk/benefit relationship to that reported in North American patients. CLINICAL TRIAL REGISTRATION ClinicalTrials.gov, No. NCT01596621; https://clinicaltrials.gov/ct2/show/NCT01596621.
Adult ; Antineoplastic Combined Chemotherapy Protocols ; Bendamustine Hydrochloride/therapeutic use* ; China ; Humans ; Lymphoma, Non-Hodgkin/drug therapy* ; Neoplasm Recurrence, Local/drug therapy* ; Prospective Studies ; Rituximab/therapeutic use*

Aim To investigate the effect of salvianolic acid B(Sal B) on the attenuation of rat hepatocyte in-jury induced by hypoxia/reoxygenation(H/R) and its possible molecular mechanism. Methods Rat hepato-cytes BRL-3A were cultured in vitro. H/R injury mod-el was established and then BRL-3A cells were pretrea-ted with Sal B. The viability of cells was measured by CCK-8 assay;the expression of ALT and AST was de-tected by microplate assay; the levels of TNF-α and IL-1β were determined by ELISA; the apoptosis was detected by flow cytometry;the protein and mRNA lev-els of SIRT1, NF-κB p65, p53, Bax and Bcl-2 were measured by Western blot and qPCR. Results H/R intervention decreased the viability and increased the apoptosis of cells;the production of ALT, AST, TNF-α and IL-1β was elevated;the protein and mRNA lev-els of SIRT1, Bcl-2 were reduced, but the levels of NF-κB p65, p53 and Bax increased. After pretreated with Sal B, the viability of cells increased while the apoptosis decreased; the expression of ALT, AST, TNF-α and IL-1β was inhibited;moreover,the protein and mRNA levels of SIRT1,Bcl-2 were enhanced,and the levels of NF-κB p65, p53 and Bax decreased sig-nificantly. Conclusion Sal B may attenuate rat hepa-tocyte injury induced by H/R via the SIRT1/NF-κB/p53 pathway.

To determine whether ginkgo biloba extract(GBE)combined with dexamethasone(DEX)plays a role in the treatment of allergic rhinitis-related olfactory dysfunction using an animal model.Six week old BALB/C mice were sensitized and challenged with ovalbumin.30 sensitized mice were divided into three groups:Group 1 was given high-dose GBE and DEX(n=10);Group 2 was given low dose GBE and DEX(n=10);Group 3 was given DEX alone(n=10).We assessed the histology of the olfactory mucosa and serum IL-4,IFN-γ,and caspase 1.A significant higher fraction of mice in group 1 could find the food pellet within300 scompared to group 3(<0.05).Caspase-1 levels improved during the second week compared with the first week in each group.IFN-γlevels were significantly lower during the second week compared with the first week(<0.05,all).IL-4 levels also were significantly lower during the second week compared with the first week in all groups except those receiving DEX alone.IFN-γ/IL-4 levels in each group were significantly lower during the second week compared with the first week(<0.05,all).In this animal model of allergic rhinitis-related olfactory dysfunction,the addition of ginkgo biloba extract to dexamethasone have a better anti-inflammatory effect,which can partly improve the therapeutic effect on olfactory dysfunction caused by allergic rhinitis.
Animals ; Cytokines ; metabolism ; Disease Models, Animal ; Glucocorticoids ; administration & dosage ; pharmacology ; Mice ; Mice, Inbred BALB C ; Plant Extracts ; administration & dosage ; pharmacology ; Rhinitis, Allergic ; drug therapy ; immunology

Objective To explore the prospective monitoring and control methods for missing reporting of health-care-associated infection(HAI)cases,analyze its implementation efficacy,provide basis for formulating targeted strategy for monitoring missing report of HAI.Methods From January 2016 to June 2017,the quality control circle (QCC)method was used to prospectively monitor HAI cases in hospitalized patients,missing reporting of HAI was controlled.Results "Information system intelligence screening+mobile messaging alerts+ HAI supervision"trinity monitoring model for avoid missing reporting of HAI cases was established,after the first round of PDCA(plan, do,check,action)cycle,missing reporting rate of HAI decreased from 79.16% before QCC to 59.75% after QCC, difference was statistically significant (χ2＝208.821,P＝0.000).Compared with missing reporting rate of HAI af-ter the first round of PDCA,missing reporting rate of HAI after the second round of PDCA dropped to 26.18%, difference was statistically significant (χ2＝200.075,P＝0.002).Conclusion Active prospective prevention and control before missing reporting of HAI can effectively avoid missing reporting of HAI cases.