Objective: To provide high-quality clinical evidence of the efficacy of Tibetan medicine Honghua Ruyi (HHRY) pills for endometriosis-associated dysmenorrhea. Methods: This study constitutes a multicenter, randomized, double-blind, placebo-controlled trial encompassing a three-menstrual cycle intervention followed by a three-menstrual cycle follow-up period. A total of 164 eligible females with endometriosis-associated dysmenorrhea were randomly divided into HHRY pills and placebo groups in a 1:1 ratio. The primary outcome included dysmenorrhea symptoms assessed using Visual Analog Scale (VAS) scores and quality of life, whereas the secondary outcome measures included the maximum VAS for non-menstrual pelvic pain, duration of pain episodes (in days), frequency and quantity of the consumption of ibuprofen sustained-release capsules (or other non-steroidal anti-inflammatory drugs), and days off work/study for staff/student due to dysmenorrhea, ovarian cyst, and/or pelvic nodule size. The safety was monitored throughout the treatment period. All the analyses were based on the intention-to-treat principle. For continuous outcomes, simple or multiple linear regressions were used to estimate the differences between the HHRY pills and placebo groups, with categorical data expressed as the number and percentage of occurrences. Differences were compared using the chi-square test or Fisher's exact test. The predefined analysis was adjusted for concomitant treatment, a variable considered to be associated with outcomes but unaffected by treatment allocation. Estimates of treatment effects were reported with 95% confidence intervals. Two-tailed P values ≤ .05 were considered statistically significant. Conclusion Positive results from this trial, upon completion would provide robust evidence for the efficacy and safety of HHRY pills in treating dysmenorrhea in patients with endometriosis.

Objective To investigate the effects of linoleic acid(LA)on the diversity and structure of intestinal flora in mice.Methods Twelve SPF grade male C57BL/6J mice at 7 weeks were randomly divided into control group(CTRL group)and linoleic acid group(LA group).One day before the linoleic acid diet was supplemented,the normal food was removed from the LA group and the mice in the LA group were fasted for one night,so that the LA diet was more acceptable to the mice in the LA group,and LA was given on the day of the experiment recording,and the feed was updated at any time to ensure that the mice could eat freely until the end of modeling.After 12 weeks of modeling,mouse feces were collected,and mixed samples were collected for every two mice feces,and then 16sRNA high-throughput sequencing was performed to analyze intestinal flora structure,Alpha and Beta diversity.Results 16sRNA high-throughput sequencing showed that LA intervention damaged the richness and diversity of intestinal flora.The results of principal component analysis showed that the composition of flora in CTRL group was different from that in LA group.At gate level,the relative abundance of Actinobacteria in LA group increased(P＜0.01).At the genus level,the relative abundance of L.Duchennei in the LA group decreased(P＜0.05),but the relative abundance of Bifidobacterium,Faecalibaculum and Erysipelotrichaceae in the LA group increased(all P＜0.01).Conclusion LA intervention could reduce the richness and diversity of intestinal flora in mice,and adjust the structure of intestinal flora.There were significant differences between beneficial bacteria and pathogenic bacte-ria in intestinal flora after LA intervention,which provided certain basis for the treatment of bioactive compounds of linoleic acid and the therapeutic adjustment of intestinal microorganisms as targets.

To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori(Hp),this study employed several strategies.First,using artificial intelligence(AI)auxiliary tools such as Pymol,I-TASSER,and ClussPro2,the molecular interactions between different antibodies and Hp urease subunit B(UreB)were analyzed.The fully humanized single-domain antibody UreBAb was identified as the primary research target.Next,the UreBAb gene sequence was optimized based on Escherichia coli codon preferences,and was inserted into expression vectors such as pET28a and pE-SUMO.The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3).Recombinant antibody proteins were prepared through IPTG induction,and its activity was detected using extracted Hp urease as the antigen.SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb,with protein yields of 0.34 mg/mL and 0.41 mg/mL,respectively.Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen,with inhibition rates of 51.27%and 74.07%,respectively.Additionally,leveraging artificial intelligence tools such as AlphaFold2,cluspro2,mCSM-AB,OSPREY,and FoldX,the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex.Subsequently,nine UreBAb evolution mutants were constructed,and their binding activities with the antigen were enhanced.Among these,the I107W mutant showed the most significant improvement,achieving a 24.95%increase compared to the wild-type UreBAb.This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.

Objective To investigate the effects of sodium-glucose cotransporter 2 inhibitor (SGLT2i)canagliflozin and epithelial sodium channel inhibitor amiloride or benzamil in combination on the bone metabolism in the rats with the nephrotic syndrome (NS) induced by doxorubicin. Methods In the 49 male SD rats selected, 7 were randomly selected as control group (NG), and 42 were built as adriamycin-induced nephropathy model by injecting adriamycin through the tail vein, and were randomly divided into model group (MG group), canagliflozin group (KG group), benzamil group (BH group), amiloride group (AL group), canagliflozin+benzamil group (KB group) and canagliflozin+amiloride group (KA group), with 7 cases in each group. Each medication group was given intragastric administration according to the body weight of rats regularly every day, NG group and MG group were given equal amount of normal saline, the course of treatment was 6 weeks. The 24-hour urinary protein (24 h-UTP) of each group was detected one day before treatment to verify the successful preparation of the model. At 6 weeks after treatment, the 24 h-UTP, urine sodium (UNa), urinary potassium (UK) levels in urine and the albumin (ALB), triglyceride (TG), cholesterol (TC), low density lipoprotein (LDL), serum calcium (SCa), sodium (SNa), potassium (SK), 25-hydroxyvitamin D (25-OH-D), alkaline phosphatase (ALP), parathyroid hormone (PTH), procollagen type Ⅰ N-terminal propeptide (PINP) and beta C-terminal cross-linked telopeptides of typeⅠ collagen (β-CTX) levels in serum were measured, respectively. Results After successful modeling, the levels of 24 h-UTP, TG, TC, LDL and SCa in the MG group were significantly increased, while the levels of ALB, 25-OH-D, ALP, PINP and PTH were significantly decreased (P < 0.05 or P < 0.01). After 6 weeks of drug intervention, the body mass of rats treated with KG alone decreased significantly (P < 0.05). Compared with the MG group, the levels of 24 h-UTP, TC, TG, LDL and SCa were significantly decreased, and the level of ALB was significantly increased in all drug groups when compared to the MG (P < 0.05 or P < 0.01). Compared with the MG group, the levels of 25-OH-D, ALP, PINP and β-CTX in the KA group were significantly increased when compared to the MG (P < 0.05 or P < 0.01). Molding and drug therapy had no impact on levels of UNa, UK, SK and SNa (P>0.05). Conclusion Calaglizin alone, benzamil alone, amiloride alone and calaglizin combined with benzamil or amiloride can improve the mass proteinuria, hypoproteinemia and hyperlipidemia in adriamycin-induced NS model rats. The combination of caraglipzin and amiloride has a significant osteogenic or osteoclastic effect, which may play a role in reducing the risk of fracture in NS rats by forming a new osteogenic/osteoclastic balance.

OBJECTIVE: To explore the therapeutic mechanism of Liushen Wan (LSW) against colitis-associated colorectal cancer (CAC) by network pharmacology. METHODS: TCMSP, BATMAN-TCM, CNKI, PubMed, Genecards, OMIM, and TTD databases were used to obtain the related targets of LSW and CAC. The common targets of LSW and CAC were obtained using Venny online website. The PPI network was constructed using Cytoscape 3.8.2 to screen the core targets of LSW in the treatment of CAC. GO and KEGG enrichment analysis were conducted using DAVID database. The therapeutic effect of LSW on CAC was evaluated in a C57BL/6J mouse model of AOM/DSS-induced CAC by observing the changes in body weight, disease activity index, colon length, and size and number of the tumor. HE staining and RT-qPCR were used to analyze the effect of LSW on inflammatory mediators. Immunohistochemistry and TUNEL staining were used to evaluate the effect of LSW on the proliferation and apoptosis of AOM/DSS-treated colon tumor cells. Immunohistochemistry and Western blotting were used to detect the effects of LSW on the expression of TLR4 proteins in CAC mice. RESULTS: Network pharmacology analysis identified 69 common targets of LSW and CAC, and 33 hub targets were screened in the PPI network. KEGG pathway enrichment analysis suggested that the effect of LSW on CAC was mediated by the Toll-like receptor signaling pathway. In the mouse model of AOM/DSS-induced CAC, LSW significantly inhibited colitis-associated tumorigenesis, reduced tumor number and tumor load (P < 0.05), obviously improved histopathological changes in the colon, downregulated the mRNA levels of proinflammatory cytokines, and inhibited the proliferation (P < 0.01) and promoted apoptosis of colon tumor cells (P < 0.001). LSW also significantly decreased TLR4 protein expression in the colon tissue (P < 0.05). CONCLUSION LSW can inhibit CAC in mice possibly by regulating the expression of TLR4 to reduce intestinal inflammation, inhibit colon tumor cell proliferation and promote their apoptosis.
Mice ; Animals ; Toll-Like Receptor 4 ; Colitis-Associated Neoplasms ; Network Pharmacology ; Mice, Inbred C57BL ; Colonic Neoplasms/pathology*

Objective:To have a better understanding of congenital leukemia by summarizing its clinical features and prognosis.Methods:This study retrospectively recruited 10 neonates with congenital leukemia treated in Beijing Children's Hospital, Capital Medical University from January 2006 to December 2018. Clinical data including clinical manifestations, laboratory examinations, treatment and follow-up were described.Results:The 10 patients consisted of six boys and four girls. In all cases, symptoms presented within 11 d after birth. The admission complaints were jaundice ( n=4), polypnea ( n=3), fever ( n=2) and rash ( n=2). Physical examinations of the 10 patients showed eight with splenomegaly, seven with hepatomegaly and seven with petechia/skin rash. All patients had significantly increased white blood cell count (from 45.8×10 9/L to 553.0×10 9/L), complicated by different degrees of anemia and thrombocytopenia. By bone marrow biopsy, two cases were diagnosed as acute lymphoblastic leukemia and the other eight cases as acute myeloid leukemia. All cases refused chemotherapy on the preliminary diagnosis. Three cases lost follow-up and six died within two months after discharge requested by their parents. One baby had spontaneous remission, but relapsed two years later. Complete remission was achieved after strict management and no relapse was reported until ten years old. Conclusions:Congenital leukemia is a severe condition with high mortality. Some cases may achieve spontaneous remission, but long-term follow-up is needed.

2,6-dimethylbenzenamine was determined as a genotoxic impurity in lidocaine hydrochloride injection, and 2-chloro-N-(2,6- dimethylphenyl) acetamide was determined as potential genotoxic impurity. An LC-MS/MS method was established to research the profiling of genotoxic impurities in active pharmaceutical ingredients (API), homemade preparation and reference preparation on column Agilent ZORBAX Eclipse Plus C18(4.6 mm250 mm，5 μm). The results show that in the homemade preparation the 2,6-dimethylbenzenamine and the 2-chloro-N-(2,6-dimethylphenyl) acetamide may be degraded under oxidation condition and alkaline condition in addition to the introduction from API preparation process. This study provides guidance for genotoxic risk assessment and prescription process optimization of lidocaine hydrochloride.

Objective: To observe the effect of the application of the "3Y" shape Multipore tape combined with transparent dressing in the maintenance of double lumen PowerPICC. Methods: Totally150 patients inserted double lumen powerPICC were randomly assigned to experimental group and control group, each group included 75 patients. when maintaining PICC, the "3Y" shape Multipore tape combined with transparent dressing were used for experimental group, while traditional method were used for control group. The endpoints were: the time for dressing change,the incidence of catheter migration,the incidence of dressing rolling or loosing,the incidence of PICC related complications. Results: The time for dressing change of experimental group (13.14±0.23) min was significant longer than control group (12.99±0.24) min (t=4.025, P<0.05) ; incidence of catheter migration of experimental group (5.3%,4/75) was significant lower than control group (18.7%, 14/75) (χ²=6.424, P=0.04). Incidence of dressing rolling or loosing of experimental group (9.3%,7/75) was significant lower than control group (22.7%,17/75)(χ²=7.028, P=0.03). Incidence of complication of experimental (1.3%, 1/75) was significant lower than control group (9.3%, 7/75) (χ²=4.754, P=0.03) . Conclusions Application of the "3Y" shape Multipore tape combined with transparent dressing in the maintenance of double lumen PowerPICC cost a little bite more time,while reduce the incidence of catheter migration and related complications, improve patients′degree of comfort, worth to be widely used.

Objective To evaluate the clinical efficacy of thyroxine replacement therapy in the treatment of pregnancy patients with subclinical hypothyroidism.Methods 80 cases of patients with subclinical hypothyroidism who were admitted to our hospital from May 2011 to May 2016 were selected, they were divided into thyroxine replacement therapy treatment group ( replacement treatment group ) and conventional treatment group two groups according to the treatment methods, 40 cases in each group.The serum TSH, FT4, TG, CHO levels, childbirth, pregnancy outcomes and complications, neonatal status of the two groups were statistically analyzed.Results The serum TSH, TG, CHO levels of the replacement treatment group were significantly lower, the FT4 level was significantly higher, the difference was statistically significant (P<0.05), the trans vaginal yield rate 70.0%(28/40) was significantly higher than the conventional treatment group 35.0%(14/40), the cesarean section rate 30.0%(12/40) was significantly lower than the conventional treatment group 65.0%( 26/40 ) , the difference was statistically significant ( P<0.05 ) , the incidences of preterm delivery, postpartum hemorrhage, preeclampsia, hypoproteinemia, fetal development restriction 2.5%(1/40), 2.5%(1/40), 5.0%(2/40), 7.5%(2/40), 0 were significantly lower than the conventional treatment group 17.5%(7/40), 15.0%(6/40), 20.0%(8/40), 17.5%(7/40), 15.0%(6/40), the difference was statistically significant (P<0.05), the neonates body weight was significantly higher than the conventional treatment group, the difference was statistically significant (P<0.05), the proportions of low birth weight infants, giant infants, premature infants, asphyxia, death 0, 2.5%(1/40), 5.0%(2/40), 7.5%(2/40), 0 were significantly lower than the conventional treatment group 15.0%(6/40), 20%(8/40), 22.5%(9/40), 27.5%(11/25), 20.0%(8/40), the difference was statistically significant (P<0.05).Conclusion The clinical efficacy of thyroxine replacement therapy in the treatment of pregnancy patients with subclinical hypothyroidism is more significant than conventional treatment .

Objective To discuss the feasibility of single hole thoracoscopy of pleural fibreboard endarterectomy surgical treatment on chronic tuberculous empyema. Methods Retrospective analysis of minimally invasive treatment of 52 cases of chronic tuberculous empyema form January 2013 to May 2016, 50 cases applied single hole thoracoscopy surgery, video-assisted mini-thoracoscopy for another 2 cases. Results There was no death, operation time 60 ~ 240 min, average 160 min, bleeding 150 ~ 2000 ml, average 350 ml, postoperative chest tube drainage time 3 ~ 21 d, average 7 d, postoperative persistent leakage in 3 patients, 3 cases of atelectasis, incisional infection in 1 case, pleural effusion in 1 case, 3 cases of arrhythmia. All the cured patients are received the corresponding treatment, the follow-up of 3 ~ 36 m, the chest CT scan show no atelectasis. Conclusion Under the condition of strict selection of indication, single hole thoracoscopy of pleural fibreboard endarterectomy in treatment of chronic tuberculous empyema is safe and feasible, so it is worthy of making further clinical promotion and application.