Objective: To assess the dynamic changes of microcirculation at acupoints in patients with primary dysmenorrhea and cold congelation and blood stasis syndrome using laser speckle blood flow imaging. Methods: Patients with primary dysmenorrhea and cold coagulation and blood stasis syndrome (primary dysmenorrhea group, n=53) and healthy female college students(control group, n=57) who met the inclusion and exclusion criteria from October 2020 to July 2022 were enrolled at Hebei University of Chinese Medicine. On the premenstrual and first day of menstruation, a laser speckle blood flow imaging system was used to measure the microcirculation blood flow perfusion on the surface of acupoints related to the conception, thoroughfare, and governor vessels, and stomach, spleen, and bladder meridians in the abdomen and lumbosacral regions. The dynamic changes in microcirculation were calculated based on the difference in average blood flow perfusion at each acupoint before and after menstruation. Receiver operating curve (ROC) analysis was used to analyze the diagnostic efficacy of dynamic changes in microcirculation on the surface of each acupoint. The microcirculation sensitization rate of acupoints was calculated. Results: Compared with the control group, the dynamic changes in microcirculation at the following acupoints in the primary dysmenorrhea group were increased (P<0.05): conception vessel (Yinjiao[CV7], Qihai[CV6], Shimen[CV5], Guanyuan[CV4]); left thoroughfare vessel (left Huangshu[KI16], left Zhongzhu[KI15], left Siman[KI14], left Qixue[KI13], left Dahe[KI12], left Henggu[KI11]); left stomach meridian (left Tianshu[ST25], left Wailing[ST26], left Qichong[ST30]); left spleen meridian (left Daheng[SP15], left Fujie[SP14]); right thoroughfare vessel (right Huangshu[KI16], right Zhongzhu[KI15], right Siman[KI14], right Qixue[KI13], right Dahe[KI12], right Henggu[KI11]); right stomach meridian (right Wailing[ST26], right Daju[ST27], right Shuidao[ST28], right Guilai[ST29], right Qichong[ST30]); and right spleen meridian (right Fujie[SP14]). The area under the ROC curve of conception vessel (Yinjiao[CV7], Qihai[CV6], Shimen[CV5], Guanyuan[CV4]), thoroughfare vessel (right Siman[KI14], left Huangshu[KI16], right Qixue[KI13], right Zhongzhu[KI15], right Dahe[KI12], left Zhongzhu[KI15], left Siman[KI14], right Huangshu[KI16], left Qixue[KI13], right Henggu[KI11], left Henggu[KI11], left Dahe[KI12]); stomach meridian (left Tianshu[ST25], right Guilai[ST29], left Wailing[ST26], right Shuidao[ST28], right Daju[ST27], right Wailing[ST26], right Qichong[ST30], left Qichong[ST30]), and spleen meridian (left Daheng[SP15], left Fujie[SP14], right Fujie[SP14]) was 0.610-0.682 (P<0.05). Compared with the control group, the sensitization rate of some acupoints in the primary dysmenorrhea group increased (P<0.05). Conclusion With the onset of menstruation, the blood flow perfusion of some acupoints in the abdomen (thoroughfare, and conception vessels, and stomach and spleen meridians) of patients with primary dysmenorrhea and cold blood coagulation and blood stasis syndrome increased, and the status of acupoints changed from a resting state to an active state. These acupoints are sensitive in patients with primary dysmenorrhea and cold blood coagulation and blood stasis syndrome and have a certain diagnostic efficacy, providing a basis for further analyzing the efficacy and mechanism of acupuncture and moxibustion to treat primary dysmenorrhea with cold blood coagulation and blood stasis syndrome.

Objective:To evaluate the clinical efficacy of acupuncture in the treatment of anxiety state in patients with primary insomnia(PI). Methods:Randomized controlled trials of acupuncture treatment for PI patients with an anxiety state in Web of Science,Cochrane Library,PubMed,Excerpta Medica Database(EMBASE),China National Knowledge Infrastructure(CNKI),Wanfang Data Knowledge Service Platform(Wanfang),and Chongqing VIP Database(VIP)were retrieved by computer.The retrieval time was from each database's inception to December 30,2022.Data extraction and evaluation were performed for the included studies.The Cochrane risk of bias assessment tool was used to assess the risk of bias in each article.Meta-analysis of valid data was performed using the RevMan 5.4 software.If the outcome indicator was a categorical variable,relative risk(RR)was used as the effect size.If it was a continuous variable,mean difference(MD)was used to calculate the effect size.Each effect size was expressed as a 95%confidence interval(CI).P<0.05 was considered to indicate a statistically significant difference. Results:A total of 18 studies were included,comprising a total of 1198 patients.The findings of the meta-analysis showed that electroacupuncture had a significant advantage in improving the Hamilton anxiety scale(HAMA)score than benzodiazepines[MD=-1.61,95%CI(-2.17,-1.06),P<0.001].Acupuncture was superior to sham acupuncture[MD=-14.90,95%CI(-20.39,-9.41),P<0.001]and benzodiazepines[MD=-3.39,95%CI(-4.67,-2.12),P<0.001]in reducing the self-rating anxiety scale(SAS)score.Acupuncture was superior to sham acupuncture in reducing the insomnia severity index(ISI)score[MD=-5.61,95%CI(-6.63,-4.89),P<0.001].Acupuncture was superior to benzodiazepines[MD=0.84,95%CI(-1.42,-0.25),P=0.005]and sham acupuncture[MD=-8.39,95%CI(-8.39,-7.86),P<0.001]in improving the Pittsburgh sleep quality index(PSQI)score.Acupuncture had a better effective rate than benzodiazepines[RR=1.16,95%CI(1.08,1.25),P<0.001]and sham acupuncture[RR=8.94,95%CI(4.63,17.25),P<0.001]in treating PI. Conclusion:Acupuncture or electroacupuncture has certain therapeutic advantages over benzodiazepines and sham acupuncture in the treatment of anxiety in PI patients.However,more high-quality randomized controlled trials are needed for further verification.

AIM To investigate the protective effects and the mechanism of the Liuwei Dihuang Pills on mouse brain microvascular endothelial(bEnd.3)cells damaged by β-Amyloid protein1-40(Aβ1-40).METHODS CCK8 method was used to detect the effects of Aβ1-40 and medicated serum of Liuwei Dihuang Pills(MSLDP)on cell activity,and to screen the appropriate concentration.bEnd.3 cells of the control group,the Aβ1-40 group,the MSLDP+Aβ1-40 group and the MSLDP group had their low density lipoprotein-associated protein 1(LRP1),receptor for advanced glycation end products(RAGE),matrix metalloproteinase-2(MMP-2),MMP-9,scaffold protein zonule protein-1(ZO-1)detected by Western blot.bEnd.3 cells assigned into the control group,the Aβ1-40 group,the FPS-ZM1(RAGE inhibitor)+Aβ1-40 group and the FPS-ZM1+Aβ1-40+MSLDP group had their expressions of RAGE,MMP-9,MMP-2 and ZO-1 detected by Western blot as well.RESULTS The cell activity of bEnd.3,was dose-dependently decreased by Aβ1-40(P<0.01),but was protected by MSLDP(P<0.05,P<0.01).And 10 μmol/L Aβ1-40 and 10%MSLDP were selected for subsequent experiments.Compared with the control group,the Aβ1-40 group displayed increased protein expressions of RAGE,MMP-2 and MMP-9(P<0.01),decreased protein expressions of LRP1,ZO-1 and BDNF(P<0.05,P<0.01),and decreased fluorescence intensities of LRP1 and ZO-1(P<0.01).Compared with the Aβ1-40 group,the MSLDP group shared decreased expressions of RAGE,MMP-2,MMP-9 proteins and RAGE fluorescence intensity(P<0.05,P<0.01),and increased expressions of LRP1,ZO-1 and BDNF proteins,and the fluorescence intensity of LRP1 and ZO-1(P<0.05,P<0.01);the Aβ1-40+FPS-ZM1 group displayed decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.05,P<0.01),and increased ZO-1 protein expression(P<0.05);and the Aβ1-40+FPS-ZM1+ MSLDP group displayed an even more decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.01),increased ZO-1 protein expression(P<0.01)due to the the combination use of FPS-ZM1 and MSLDP.CONCLUSION Liuwei Dihuang Pills can protect the tight junction of bEnd.3 injured by Aβ1-40 and neurovascular units from Alzheimer's disease by alleviating the dysfunction of the blood-brain barrier via RAGE-mediated MMP-2/MMP-9 pathway inhibition.

ObjectiveTranscription factor NFE2 was observed abnormal expression in myeloproliferative neoplasm (MPN) patients. However, how NFE2 is transcriptionally regulated remains ambiguous. This study aims to explore the elements and molecular mechanisms involved in the transcriptional regulation of NFE2. MethodsActive enhancers were predicted by public NGS data and conformed experimentally via dual luciferase reporter assay. After that, PRO-seq and GRO-seq data was used to detect enhancer RNAs transcribed from these enhancers. RACE was utilized to clone the full length enhancer RNA (eRNA) transcripts, and RT-qPCR was used to measure their expression in different leukemia cell lines as well as the transcript levels during induced differentiation. Finally, to investigate the molecular function of the eRNA, overexpression and knockdown of the eRNA via lentivirus system was performed in K562 cells. ResultsWe identified three enhancers regulating NFE2 transcription, which located at -3.6k, -6.2k and +6.3k from NFE2 transcription start site (TSS) respectively. At the -3.6k enhancer, we cloned an eRNA transcript and characterized that as a lncRNA which was expressed and located in the nucleus in three types of leukemia cell lines. When this lncRNA was overexpressed, expression of NFE2 was upregulated and decreases of K562 cell proliferation and migration ability were observed. While knocking down of this lncRNA, the level of NFE2 decreases correspondingly and the proliferation ability of K562 cells increases accordingly. ConclusionWe identified an enhancer lncRNA that regulates NFE2 transcription positively and suppresses K562 cell proliferation.

Objective We aimed to observe the differences in the effects of different acupuncture technique on the endometrium of mice with simple endometrial hyperplasia model and to explore the potential mechanisms. Methods According to the random number tables,32 female C57BL/6J mice were divided into a blank control group,a model group,a quick needle group and a retaining needle group,with 8 mice in each group. A mouse model of simple endometrial hyperplasia was established using bilateral ovariectomy combined with estrogen loading. In the quick needle group,mice were punctured at the bilateral for "Yinbai"(SP1) points and withdrawn immediately,with the treatmeat performed once every other day for a total of 12 times. In the retaining needle group,mice were punctured at the bilateral "Yinbai"(SP1) points and the needles were retained for 15 min each time,with the treatment also performed once every other day for a total of 12 times. After the intervention,samples were collected. HE staining was used to observe morphological changes in the mouse uterine tissue;ELISA was used to detect serum estradiol level;flow cytometry was used to detect the ratio of M1 and M2 macrophages(M1/M2) and immunohistochemical method was used to measure the expression of CD86 and CD206 in uterine tissue;and Western blotting was used to detect the expression of interleukin-13 (IL-13) and interferon-γ(IFN-γ) in uterine tissue. Results The endometrium of mice in the model group showed simple hyperplasia. Compared with the blank control group,the endometrium of the model group was thickened (P<0.01);the level of estradiol in the serum was increased (P<0.01);M1/M2 in uterine tissues was decreased (P<0.01),the expression of CD86 was decreased (P<0.01),and the positive expression of CD206 was increased (P<0.01);and the level of IFN-γ protein expression in uterine tissues was decreased (P<0.01),and the expression of IL-13 protein was increased (P<0.01). Compared with the model group,the endometrial thicknesses of the quick needle group and the retaining needle group were reduced (P<0.05),the levels of estradiol in serum were reduced (P<0.05),M1/M2 in uterine tissues increased (P<0.01),and the reduction of CD206 positive expression,and IL-13 protein expression reduced (P<0.01);the level of CD86 positive expression,IFN-γ protein expression increased (P<0.01). Compared with the quick needle group,IL-13 protein expression increased in the retaining needle group (P<0.01).Conclusion Both quick needle and retaining needle may be through the regulation of the expression of IFN-γ and IL-13,thus prompting the polarization of macrophages from M2 to M1 type,inhibiting the pro-cell proliferative ability and tissue repair ability of M2 type macrophages,thus reducing the degree of endometrial hyperplasia,and the quick needle group was superior to the retaining needle group in regulating the expression of IL-13.

Objective:To compare and evaluate recurrence patterns after robotic-assisted gastrectomy (RAG) versus laparoscopic-assisted gastrectomy (LAG).Methods:This was a retrospective cohort study of 2915 consecutive patients with gastric adenocarcinoma confirmed by postoperative histology as T1-4aN0-3M0, who had undergone minimally invasive radical gastrectomy at four large gastric cancer treatment centers (Fujian Medical University Union Hospital: 1426 patients; the First Affiliated Hospital, Nanchang University: 1108; Tianjin Medical University Cancer Institute and Hospital: 196; and First Affiliated Hospital of Xi'an Jiaotong University: 185 cases) between 1 January 2015 and 30 June 2019. 930 patients had undergone RAG (RAG group) and 1985 had undergone LAG (LAG group). We assessed the following characteristics: age, sex, body mass index, American Society of Anesthesiologists score, comorbidities, tumor size, extent of surgery, extent of lymph node dissection, pT, pN, year of surgery, and adjuvant chemotherapy, after propensity score matching (1:1). There were no significant differences in baseline clinical characteristics between the two groups formed by propensity score matching (837 in each group) (all P>0.05). The 3-year recurrence-free survival (RFS), recurrence pattern, and conditional RFS were compared. Results:We detected no significant differences in the overall recurrence rate at 3 years (128/837 [15.3%] vs. 141/837 [16.8%], P=0.387) or time to recurrence (15.7±8.1 months vs. 16.4±8.4 months, P=0.449) between the RAG and LAG groups. Peritoneal recurrence was the most common type of recurrence in both groups (55 [6.6%] vs. 69 [8.2%], P=0.524). The difference in 3-year RFS between the RAG and LAG groups was not statistically significant (83.2% vs. 82.5%, P=0.781). We found that age > 60 years, total gastrectomy, and worse pT stage and pN stage were independent risk factors for recurrence in the study patients (all P<0.05), whereas the surgical procedure (RAG or LAG) was not an independent risk factor for RFS ( P=0.242). The 3-year conditional RFS at various time points was comparable between the two groups (1 year postoperatively: 84.6% vs. 84.7%, P=0.793; 3 years postoperatively: 91.5% vs. 94.9%, P=0.647). Conclusions:In this multicenter study of patients with locally resectable gastric cancer, we demonstrated that RAG performed by surgeons at large gastric cancer centers is not inferior to LAG in 3-year recurrence rate or recurrence patterns.

Objective:To compare and evaluate recurrence patterns after robotic-assisted gastrectomy (RAG) versus laparoscopic-assisted gastrectomy (LAG).Methods:This was a retrospective cohort study of 2915 consecutive patients with gastric adenocarcinoma confirmed by postoperative histology as T1-4aN0-3M0, who had undergone minimally invasive radical gastrectomy at four large gastric cancer treatment centers (Fujian Medical University Union Hospital: 1426 patients; the First Affiliated Hospital, Nanchang University: 1108; Tianjin Medical University Cancer Institute and Hospital: 196; and First Affiliated Hospital of Xi'an Jiaotong University: 185 cases) between 1 January 2015 and 30 June 2019. 930 patients had undergone RAG (RAG group) and 1985 had undergone LAG (LAG group). We assessed the following characteristics: age, sex, body mass index, American Society of Anesthesiologists score, comorbidities, tumor size, extent of surgery, extent of lymph node dissection, pT, pN, year of surgery, and adjuvant chemotherapy, after propensity score matching (1:1). There were no significant differences in baseline clinical characteristics between the two groups formed by propensity score matching (837 in each group) (all P>0.05). The 3-year recurrence-free survival (RFS), recurrence pattern, and conditional RFS were compared. Results:We detected no significant differences in the overall recurrence rate at 3 years (128/837 [15.3%] vs. 141/837 [16.8%], P=0.387) or time to recurrence (15.7±8.1 months vs. 16.4±8.4 months, P=0.449) between the RAG and LAG groups. Peritoneal recurrence was the most common type of recurrence in both groups (55 [6.6%] vs. 69 [8.2%], P=0.524). The difference in 3-year RFS between the RAG and LAG groups was not statistically significant (83.2% vs. 82.5%, P=0.781). We found that age > 60 years, total gastrectomy, and worse pT stage and pN stage were independent risk factors for recurrence in the study patients (all P<0.05), whereas the surgical procedure (RAG or LAG) was not an independent risk factor for RFS ( P=0.242). The 3-year conditional RFS at various time points was comparable between the two groups (1 year postoperatively: 84.6% vs. 84.7%, P=0.793; 3 years postoperatively: 91.5% vs. 94.9%, P=0.647). Conclusions:In this multicenter study of patients with locally resectable gastric cancer, we demonstrated that RAG performed by surgeons at large gastric cancer centers is not inferior to LAG in 3-year recurrence rate or recurrence patterns.

Sucrose synthase plays a crucial role in the plant sugar metabolism pathway by catalyzing the production of uridine diphosphate (UDP)-glucose, which serves as a bioactive glycosyl donor for various metabolic processes. In this study, a sucrose synthase gene named CtSus was cloned from Cistanche tubulosa, a traditional Chinese medicine known for its rich content of diverse glycosides, based on transcriptome analysis results. The open reading frame of CtSus was 2 418 bp long encoding 805 amino acids. Sequence analysis revealed conserved domains associated with the plant sucrose synthase family at both the N-terminal and C-terminal regions of CtSus protein. Phylogenetic analysis demonstrated that CtSus shares a close evolutionary relationship with sucrose synthases from other plants within the same order. Functional identification of CtSus was performed through whole-cell catalysis coupling with UGT71BD1, a characterized glycosyltransferase enzyme. The results showed that the introduction of CtSus significantly enhanced the conversion rate of glycosylation catalyzed by UGT71BD1. The soluble expression of CtSus in Escherichia coli was further achieved using the pCold^TM TF expression vector. In vitro enzymatic assay indicated the activity of CtSus to catalyze the formation of UDP-glucose in the presence of sucrose and UDP. Real-time quantitative-polymerase chain reaction results showed that the expression patterns of CtSus gene in different parts of C. tubulosa and the suspension cell cultures of C. tubulosa under drought stress correlated with the accumulation patterns observed for phenylethanol glycosides, respectively. Furthermore, key amino acids and their interactions between enzyme and substrate were explored based on protein structure prediction and molecular docking results. Overall, our findings identify a sucrose synthase CtSus responsible for the supply of the active glycosyl donor UDP-glucose during the biosynthesis of glycoside products in C. tubulosa and have also provided a gene element that can be utilized in engineering strain construction for glycoside products production.

Objective: To explore the characteristics of plasma Epstein-Barr virus (EBV) DNA in primary infection in pediatric cases. Methods: The laboratory and clinical data of 571 children diagnosed with EBV primary infection in Children's Hospital of Fudan University during September 1^st, 2017 to September 30^th, 2018 were retrospectively analyzed. According to the results of plasma EBV DNA, they were divided into positive group and negative group. According to the EBV DNA, they were devided into high plasma virol load group and low plasma virol load group. The Chi-square test, Wilcoxon rank sum test were used to compare the differences between groups. Results: Among the 571 children with EBV primary infection, 334 were males and 237 were females. The age of first diagnosis was 3.8 (2.2, 5.7) years. There were 255 cases in positive group and 316 cases in negative group. The percentage of cases with fever,hepatomegaly and (or) splenomegaly, elevated transaminase in the positive group were higher than those in the negative group (235 cases (92.2%) vs. 255 cases (80.7%), χ²=15.22, P<0.001; 169 cases (66.3%) vs. 85 cases (26.9%), χ²=96.80, P<0.001; and 144 cases (56.5%) vs. 120 cases (38.0%), χ²=18.27, P<0.001; respectively).In the positive group, 70 cases were followed up for 46 (27, 106) days, 68 cases (97.1%) turned negative within 28 days, with the exception of 2 cases (2.9%) developed chronic active EBV infection by follow-up revision.There were 218 cases in high plasma viral DNA copies group and 37 cases in low copies group. More cases presented with elevated transaminases in the high plasma viral DNA copies group than those in the low group (75.7% (28/37) vs. 56.0%(116/207), χ²=5.00, P=0.025).Both the positive rate of EBV DNA in peripheral blood leukocytes (84.2% (266/316) vs. 44.7% (255/571), χ²=76.26, P<0.001) and the copies of EBV DNA (7.0×10⁷ (1.3×10⁷, 3.0×10⁸) vs. 3.1×10⁶ (1.6×10⁶, 6.1×10⁶) copies /L, Z=15.23, P<0.001) were higher than that of plasma. Conclusions: In immunocompetent pediatric cases diagnosed as EBV primary infection, cases with positive plasma EBV DNA were prone to have fever, hepatomegaly and (or) splenomegaly, and elevated transaminase than those with negative plasma viral DNA. The plasma EBV DNA usually turns negative within 28 days after initial diagnosis.Most cases with high viral load in plasma showed elevated aminotransferase.
Female ; Male ; Humans ; Child ; DNA, Viral ; Herpesvirus 4, Human ; Epstein-Barr Virus Infections ; Hepatomegaly ; Retrospective Studies ; Splenomegaly ; Fever ; Transaminases

Objective: To observe the effects of different frequencies of pricking-bloodletting at auricular points plus auricular point sticking therapy on the levels of the serum immunoglobulin G (IgG), immunoglobulin M (IgM), testosterone (T), and insulin-like growth factor-1 (IGF-1) in patients with acne vulgaris (AV). Methods: Ninety patients with AV were randomly assigned to treatment group 1, treatment group 2, and treatment group 3 according to the random number remainder grouping method, with 30 cases in each group. All three groups received pricking-bloodletting at auricular points plus auricular point sticking therapy. Treatment group 1 was treated once a week, treatment group 2 was treated twice a week, and treatment group 3 was treated 3 times a week. Four-week treatment was taken as 1 treatment course, and 3 treatment courses were observed. On the day before the start of the study and on the day next to the end of each course, the global acne grading system (GAGS) score was recorded, and 3 mL of blood from the median cubital vein was collected to test the serum levels of IgG, IgM, T, and IGF-1. After 3 courses of treatment, the efficacy index was calculated according to the GAGS score, and the serum indicators in patients with effective treatment (efficacy index ≥20％) and complete sampling were analyzed. Results: A total of 60 patients were included in the final blood indicator analysis. After 3 courses of treatment, the intra-group comparisons showed that the serum IgG level increased significantly in patients in treatment group 1 (P<0.01); the serum T level decreased significantly in the female patients in treatment group 2 (P<0.05); the IGF-1 level significantly decreased in the patients in all three groups (P<0.05 or P<0.01). There were no significant differences in the serum levels of IgG, IgM, T, and IGF-1 among the three groups at each time point (P>0.05). Conclusion: Pricking-bloodletting at auricular points plus auricular point sticking therapy can affect the levels of serum IgG, T, and IGF-1 in AV patients. The level of the serum IGF-1 can be reduced by treatment once a week, twice a week, or 3 times a week. Treatment once a week can increase the patients' serum IgG level, and treatment twice a week can significantly decrease the serum T level in female patients. Reducing the serum IGF-1 level may be one of the action mechanisms of pricking-bloodletting at auricular points plus auricular point sticking therapy in treating AV.