OBJECTIVE To explore the changes and correlations of color， components and anti-tumor activity of Gelsemium elegans during sand-frying processing， so as to provide a reference for the toxicity-attenuating and efficacy-preserving processing as well as quality control of sand-fried G. elegans decoction pieces.METHODS Raw G.elegans and its processed products prepared by sand-frying for 15， 30， 75， 105 and 120 s were taken as test samples. The chromatic values， contents of total alkaloids and seven main active components （gelsemine， humantenine， koumine， gelsenicine， etc.） were determined. Their in vitro anti-tumor activity was investigated. The correlations between color and component contents， as well as between total alkaloid content and anti-tumor activity were analyzed.RESULTS The color of G.elegans samples generally darkened after sand-frying， and the total color difference （Δ E * ） of samples sand-fried for 75 s was greater than 20. At the initial stage of sand-frying （15-30 s）， the total alkaloid content in processed products decreased sharply compared with the raw material； at the later-stage of sand-frying （75-105 s）， the declining rate of total alkaloid content slowed down and tended to be stable. The contents of gelsenicine and humantenidine in processed products dropped drastically at 30 s of sand-frying， while the contents of koumine and sempervirine decreased substantially （by about 80%） after 75 s of sand-frying and then gradually tended to be stable. The anti-HepG2 cell proliferation activity of each processed product decreased with the extension of frying time. Correlation analysis demonstrated that color brightness and yellow-blue chromaticity were significantly positively correlated with the contents of total alkaloids and all determined active components except gelsemine （ P ＜0.05 or P ＜0.01）； red-green chromaticity was significantly positively correlated with the contents of koumine， sempervirine， gelsemivirine and humantenidine （ P ＜0.05 or P ＜0.01）； Δ E * was significantly negatively correlated with the contents of total alkaloids and all determined active components except gelsemine. Besides， total alkaloid content was significantly negatively correlated with the half maximal inhibitory concentration of the samples on HepG2 cells （ P ＜0.05 or P ＜0.01）.CONCLUSIONS With the prolongation of sand-frying time， the color of sand-fried G.elegans decoction pieces gradually turned brownish-brown， the contents of seven active components and total alkaloids decreased progressively， and its anti-HepG2 cell activity showed an overall downward trend. Sand-frying for 75 s serves as a critical turning-point for the above-mentioned changes.

OBJECTIVE: To observe the effects of point-moxibustion with Zhuang medicinal thread on differentially expressed genes (DEGs) in the dorsal root ganglion (DRG), tissue morphology, and the expression of Fc epsilon RI (FcεRI) pathway proteins spleen tyrosine kinase (Syk) and membrane spanning 4-domain A2 (MS4A2) in rat model of postherpetic neuralgia (PHN), and to explore the potential mechanism by which this therapy alleviates pain sensitization. METHODS: Thirty-nine male Sprague-Dawley (SD) rats were randomly divided into a control group, a model group, and a moxibustion group, with 13 rats in each group. The PHN model was established in the model and moxibustion groups by intraperitoneal injection of resiniferatoxin. In the moxibustion group, bilateral L₄-L₆ "Jiaji" (EX-B2) points were treated with point-moxibustion with Zhuang medicinal thread from day 7 post-modeling, with two cones per acupoint per session, every other day for a total of 10 sessions. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 1 day before modeling and on days 1, 4, 7, 13, 19, and 25 after modeling. After intervention, HE staining was used to observe DRG morphology. RNA sequencing was performed to analyze DEGs in DRG and conduct bioinformatics analysis. The expression of Syk and MS4A2 mRNA and proteins in the FcεRI pathway in DRG was detected by quantitative PCR and Western blot. RESULTS: Compared with the control group, the model group exhibited decreased MWT (P<0.05) and increased TWL (P<0.05); histopathological analysis revealed neuronal atrophy, nuclear displacement, and intracellular vacuoles, with a slightly loose arrangement; the RNA-Seq identified 3,207 DEGs (1,997 upregulated and 1,210 downregulated); the mRNA and protein expression levels of Syk and MS4A2 were significantly increased (P<0.01). Compared with the model group, the moxibustion group showed increased MWT (P<0.05) and decreased TWL (P<0.05), with relatively normal neuronal morphology; the RNA-Seq identified 426 DEGs (250 upregulated and 176 downregulated); the mRNA and protein expression levels of Syk and MS4A2 were significantly decreased (P<0.05). Venn diagram analysis identified 156 DEGs that showed a reversal in expression trends after treatment, including Syk and MS4A2, which are associated with pain sensitization. KEGG pathway analysis indicated that these DEGs were primarily enriched in the FcεRI pathway. CONCLUSION Point-moxibustion with Zhuang medicinal thread could alleviate pain sensitization in PHN rats, possibly by inhibiting the FcεRI signaling pathway and downregulating the expression of Syk and MS4A2.
Animals ; Rats, Sprague-Dawley ; Male ; Rats ; Moxibustion ; Neuralgia, Postherpetic/physiopathology* ; Syk Kinase/metabolism* ; Acupuncture Points ; Humans ; Ganglia, Spinal/metabolism* ; Signal Transduction

Objective To evaluate the effects of low-dose intrathecal morphine(ITM)combined with liposomal bupivacaine adductor canal block(LB-ACB)on postoperative analgesia and opioid-sparing efficacy in patients undergoing total knee arthroplasty(TKA).Methods In this randomized,double-blind,controlled trial,80 TKA patients were allocated to either an intervention group(ITM 0.1 mg+LB-ACB,n=40)or a control group(intrathecal saline+LB-ACB,n=40).Primary outcomes included resting/movement visual analog scale(VAS)scores at 6,12,24,48,and 72 hours postoperatively,48-hour morphine consumption,time to first rescue analgesia,and incidence of complications.Results(1)The intervention group showed significantly lower resting and movement VAS scores at 6,12,24,and 48 hours postoperatively compared with controls(all P<0.05),except at 72 hours(P>0.05).(2)The intervention group had a significant reduction in 48-hour morphine consumption(4.58±1.0 mg vs.9.34±4.8 mg,P=0.027),a significantly lower rescue analgesia rate(15.0%vs.47.5%,P=0.002),and a significantly prolonged time to first rescue analgesia(48.8±7.5 h vs.14.5±5.5 h,P<0.001).(3)The intervention group demonstrated a significant decrease in the incidence of nausea(from 15.0%to 35.0%,P=0.039)and vomiting(from 10.0%to 27.5%,P=0.045),but no significant differences were observed in the incidences of pruritus,urinary retention,or motor block(all P>0.05).Conclusion Low-dose ITM(0.1 mg)combined with LB-ACB significantly enhances early postoperative analgesia,reduces opioid consumption,and decreases nausea/vomiting risk,without increasing the risks of other complications.This regimen aligns with enhanced recovery after surgery(ERAS)principles.

Objective To investigate the correlation between plasma vitamin D levels and a novel inflam-matory marker,the systemic immune-inflammatory index(SII),in patients with type 2 diabetes.Methods This study adopted a cross-sectional design,in which patients diagnosed with type 2 diabetes who were admitted to the First Affiliated Hospital of Guangxi Medical University were enrolled as study participants.Data on demographic characteristics,medical history,physical examination findings,and laboratory test results were systematically collected.Participants were categorized into three groups based on their serum vitamin D levels:deficient,insuffi-cient,and sufficient.The relationship between vitamin D levels and the SII was evaluated using a multivariate linear regression model.Additionally,a restricted cubic spline model was employed to assess the nonlinear dose-response association between vitamin D levels and SII.Results This study enrolled a total of 5,716 patients with type 2 diabetes.A statistically significant difference in the SII was observed across groups with varying vitamin D levels(P<0.05),with the highest SII value found in the vitamin D-deficient group.Multivariate linear regression analysis revealed that,after adjusting for potential confounding factors including gender,age,season of blood collection,body mass index,hypertension,dyslipidemia,and chronic kidney disease,vitamin D levels were negatively associ-ated with SII(β=-2.68,95%CI:-3.56 to-1.81,P<0.001).Compared with the vitamin D-deficient group,the vitamin D-sufficient group exhibited significantly lower SII levels(β=-78.42,95%CI:-137.90 to-18.93,P=0.01).Furthermore,the restricted cubic spline model indicated a nonlinear dose-response relationship between vita-min D levels and SII(P<0.001).Conclusion There is a significant inverse correlation between plasma vitamin D levels and the SII in patients with type 2 diabetes.

Objective To investigate the correlation between plasma vitamin D levels and a novel inflam-matory marker,the systemic immune-inflammatory index(SII),in patients with type 2 diabetes.Methods This study adopted a cross-sectional design,in which patients diagnosed with type 2 diabetes who were admitted to the First Affiliated Hospital of Guangxi Medical University were enrolled as study participants.Data on demographic characteristics,medical history,physical examination findings,and laboratory test results were systematically collected.Participants were categorized into three groups based on their serum vitamin D levels:deficient,insuffi-cient,and sufficient.The relationship between vitamin D levels and the SII was evaluated using a multivariate linear regression model.Additionally,a restricted cubic spline model was employed to assess the nonlinear dose-response association between vitamin D levels and SII.Results This study enrolled a total of 5,716 patients with type 2 diabetes.A statistically significant difference in the SII was observed across groups with varying vitamin D levels(P<0.05),with the highest SII value found in the vitamin D-deficient group.Multivariate linear regression analysis revealed that,after adjusting for potential confounding factors including gender,age,season of blood collection,body mass index,hypertension,dyslipidemia,and chronic kidney disease,vitamin D levels were negatively associ-ated with SII(β=-2.68,95%CI:-3.56 to-1.81,P<0.001).Compared with the vitamin D-deficient group,the vitamin D-sufficient group exhibited significantly lower SII levels(β=-78.42,95%CI:-137.90 to-18.93,P=0.01).Furthermore,the restricted cubic spline model indicated a nonlinear dose-response relationship between vita-min D levels and SII(P<0.001).Conclusion There is a significant inverse correlation between plasma vitamin D levels and the SII in patients with type 2 diabetes.

Objective To evaluate the effects of low-dose intrathecal morphine(ITM)combined with liposomal bupivacaine adductor canal block(LB-ACB)on postoperative analgesia and opioid-sparing efficacy in patients undergoing total knee arthroplasty(TKA).Methods In this randomized,double-blind,controlled trial,80 TKA patients were allocated to either an intervention group(ITM 0.1 mg+LB-ACB,n=40)or a control group(intrathecal saline+LB-ACB,n=40).Primary outcomes included resting/movement visual analog scale(VAS)scores at 6,12,24,48,and 72 hours postoperatively,48-hour morphine consumption,time to first rescue analgesia,and incidence of complications.Results(1)The intervention group showed significantly lower resting and movement VAS scores at 6,12,24,and 48 hours postoperatively compared with controls(all P<0.05),except at 72 hours(P>0.05).(2)The intervention group had a significant reduction in 48-hour morphine consumption(4.58±1.0 mg vs.9.34±4.8 mg,P=0.027),a significantly lower rescue analgesia rate(15.0%vs.47.5%,P=0.002),and a significantly prolonged time to first rescue analgesia(48.8±7.5 h vs.14.5±5.5 h,P<0.001).(3)The intervention group demonstrated a significant decrease in the incidence of nausea(from 15.0%to 35.0%,P=0.039)and vomiting(from 10.0%to 27.5%,P=0.045),but no significant differences were observed in the incidences of pruritus,urinary retention,or motor block(all P>0.05).Conclusion Low-dose ITM(0.1 mg)combined with LB-ACB significantly enhances early postoperative analgesia,reduces opioid consumption,and decreases nausea/vomiting risk,without increasing the risks of other complications.This regimen aligns with enhanced recovery after surgery(ERAS)principles.

Objective To preliminarily investigate the association between changes in intestinal microbiota and oral microbiota of allogeneic hematopoietic stem cell transplantation(allo-HSCT)patients and early gastrointestinal acute graft versus host disease(aGVHD),and try to explore potentially effective biomarkers and provide theoretical basis for early prediction and intervention of gastrointestinal aGVHD.Methods Ten acute leukemia patients who developed gastrointestinal aGVHD within 1 month after receiving allo-HSCT in Department of Hematology of Sichuan Provincial People's Hospital from September 2021 to June 2023 were enrolled,and their fecal samples and saliva samples before and after the aGVHD were collected.16S rRNA sequencing analysis was applied for the differential changes in intestinal and oral microbiota before and after the development of early gastrointestinal aGVHD.Results ① A decrease in Bacteroides spp.and an increase in Enterococcus spp.and Enterobacteriaceae spp.in the intestinal microbiota were positively correlated with the occurrence of early upper gastrointestinal aGVHD(P＜0.05),whereas no significant difference was observed in the overall microbial diversity of the oral microbiota(P＞0.05).② LEfSe analysis of the intestinal microbiota before and after gastrointestinal aGVHD revealed an increase in Klebsiella spp.and Enterococcus spp.and a decrease in Escherichia coli;In the oral microbiota,LEfSe analysis revealed 10 microbial markers with significant difference,of which Gamma proteobacteria was the most significant.③ The difference in β-diversity of the intestinal microbiota was significant(P=0.03),whereas there was no significant difference in the α-and β-diversity of the oral microbiota.Conclusion Significant differences are found in intestinal microbiota before and after the occurrence of early gastrointestinal aGVHD in patients after Allo-HSCT,and the occurrence may have a correlation with the chang in oral microbiota.

Objective:To explore the efficacy and safety of cryopreservation-free integrated autologous hematopoietic stem cell transplantation (HSCT) model for patients with multiple myeloma.Methods:A total of 96 patients with newly diagnosed multiple myeloma (NDMM) between July 31, 2020, and December 31, 2022, were retrospectively analyzed, of which 41 patients in the observation group received integrated non-cryopreserved transplantation mode. After hematopoietic stem cells were mobilized and collected, melphalan was started immediately for pre-transplant conditioning, and non-cryopreserved grafts from the medical blood transfusion refrigerator were directly injected intravenously into the patient within 24-48 h after the melphalan conditioning. The control group consisted of 55 patients who received traditional transplantation mode. After hematopoietic stem cells were collected, stem cell cryopreservation was performed in liquid nitrogen, and then the transplant plans were started at the right time. All patients received mobilization of autologous hematopoietic stem cells using the G-CSF combined with the plerixafor.Results:① A total of 34 patients (82.9% ) with VGPR plus CR in the observation group were significantly higher than 33 patients (60.0% ) in the control group ( P=0.016). ②Compared with the control group, the incidence of grade 1 oral mucosal inflammation was higher in the observation group ( P<0.001) ; however, the incidence of grades 2 and 3 oral mucosal inflammation was lower ( P=0.004, P=0.048), and neither group experienced grade 4 or above oral mucosal inflammation. The incidence of grade 1 diarrhea was higher in the observation group ( P=0.002), whereas the incidence of grade 3 diarrhea was lower ( P=0.007). No statistically significant difference was observed in the incidence of grade 4 diarrhea ( P=0.506), and neither group experienced grade 5 diarrhea. ③ The incidence of bacterial infection in the observation group was lower than that in the control group (34.1% vs 65.5%, P=0.002), whereas no statistically significant difference was observed in the incidence of fungal infection (29.3% vs 31.4%, P=0.863) and viral infection (4.88% vs 3.64%, P=0.831). ④No statistically significant difference was observed in the implantation time of granulocytes and platelets between the observation and control groups [10 (8-20) days vs 11 (8-17) days, P=0.501; 13 (10-21) days vs 15 (10-20) days, P=0.245]. ⑤ All patients did not receive lenalidomide treatment 100 days post-transplantation. At 30 days post-transplantation, the CTL, NK, and Th cell counts in the observation group were lower than those in the control group ( P<0.001, P=0.002, P=0.049), and the NKT cell counts were higher than those in the control group ( P=0.024). At 100 days post-transplantation, the CTL, NKT, and Th cell counts in the observation group were higher than those in the control group ( P=0.025, P=0.011, P=0.007), and no statistically significant difference in NK cell counts was observed between the two groups ( P=0.396). ⑥ The median follow-up was 18 (4-33) months. The overall 2-year survival rates of the observation and control groups post-transplantation were 91.5% and 78.2%, respectively ( P=0.337). The recurrence-free survival rates were 85.3% and 77.6%, respectively ( P=0.386), and the cumulative recurrence rates were 9.8% and 16.9%, respectively ( P=0.373) . Conclusion:In NDMM, the cryopreservation-free integrated autologous HSCT model can achieve similar therapeutic effects as traditional transplantation models, with lower rates of severe mucosal inflammation and infection compared with traditional transplantation models.

OBJECTIVE To study the best extraction process of β-asarone from Acori Tatarinowii Rhizoma by ethanol heating reflux method, and to explore the antioxidant activity of different segments. METHODS With β-asarone from Acori Tatarinowii Rhizoma as the evaluation index to optimize the extraction method. On the basis of a single factor experiment, the effects of ethanol concentration, solid-liquid ratio and extraction time on the extraction amount of β-asarone from Acori Tatarinowii Rhizoma were investigated by orthogonal design and response surface methodology. After the optimal extraction process was determined, the antioxidant activities of different segments were studied. RESULTS The optimum extraction process of β-asarone from Acori Tatarinowii Rhizoma was as follows: ethanol concentration was 95%, solid-liquid ratio was 1∶20 g·mL–1 and extraction time was 2.5 h. Under these conditions, the extraction amount of β-asarone from Acori Tatarinowii Rhizoma was 0.918 7 mg·g–1. The results of in vitro antioxidant activity showed that the order of antioxidant capacity was ethyl acetate>petroleum ether>ethanol>n-butanol. Among them, the ethyl acetate fraction had the strongest antioxidant activity, with good ability to scavenge DPPH and ABTS free radicals, and had certain reduction ability. CONCLUSION The optimized method is stable, reliable and simple, which can be used for extraction and antioxidant activity determination of β-asarone from Acori Tatarinowii Rhizoma, and provides a basis for the further development of Acori Tatarinowii Rhizoma.

Objective Based on the secreted frizzled-related protein 2(SFRP2)-Wnt/β-catenin signaling pathway,this study explored the effect and mechanism of Cuiru Keli(CRKL)in the treatment of postpartum hypogalactia.Methods A rat model of postpartum hypogalactia was established by gavaging 2 mL of 1.6 mg/mL bromocriptine mesylate to female rats on the third day after delivery.Female rats with a delivery time difference of less than 48 hours were selected and randomly assigned to 7 groups,including a normal group(without any modeling or medication),a model group,a CRKL low-dose group of model group model rats receiving CRKL at the dose of 3 g/kg,a CRKL medium-dose group of model rats receiving CRKL at the dose of 6 g/kg,a CRKL high-dose group of model rats receiving CRKL at the dose of 9 g/kg,a positive drug group of model rats receiving domperidone at the dose of 3 mg/kg,and a negative control(NC)group of model rats receiving normal saline.Each group contained 6 rats.Except for the normal and model groups,the remaining 5 groups were continuously administered with the respective intervention drugs at the specified doses by gavage once a day for 10 days.Changes in the total litter mass of the offspring in the 7 groups within 10 days were measured,and HE staining was performed to identify pathological changes in the mammary tissue(MT).Six groups of rats(excluding the positive control group)were used to observe the pathological changes of eosinophils in pituitary tissue.ELISA was performed to determine the content of prolactin(PRL)in serum,immunohistochemical staining was used to determine the expression of prolactin receptor(PRLR)in MT,and RT-qPCR was used to determine the mRNA expression of genes related to lactation in MT.Network pharmacology and molecular docking were used to study the therapeutic effect and mechanism of CRKL on postpartum hypogalactia,particularly whether it acted through the SFRP2-Wnt/β-catenin signaling pathway.The mechanism of CRKL treatment was further validated by detecting mRNA(RT-qPCR)and protein expression(Western blot)of related pathway genes.Cell experiments were conducted using primary culture rat mammary epithelial cells(RMEC)from rat MT.RMEC were divided into four groups,including a normal group(primary culture RMEC,untreated),SFRP2 overexpression group(primary cultured RMEC treated with SFRP2 overexpression vector),SFRP2 overexpression+CRKL group(receiving treatment for SFRP2 overexpression group plus 10% drug-containing serum),and negative control group(primary culture RMEC treated with empty vector).The effect of CRKL on the expression of lactation-related genes FASN,CSN2,and GLUT1 mRNA after SFRP2 overexpression was detected by RT-qPCR.Results In this study,CRKL was administered at a dose of 3 g/kg in the CRKL low-dose group,6 g/kg in the medium-dose group,and 9 g/kg in the high-dose group(P<0.05 or P<0.01).Compared with the model group,CRKL at all doses significantly increased the total litter weight gain of the offsprings within 10 days(P<0.05 or P<0.01),and effectively increased lactation(P<0.01),the area of mammary lobules,and the size and filling of acinar cavities.CRKL at all doses also increased the number of eosinophils that secreted PRL in the pituitary gland of the postpartum hypogalactia rat model,and increased the content of PRL in the serum(P<0.05 or P<0.01).CRKL promoted the secretion and expression of PRL in postpartum hypogalactic model rats.In addition,it significantly promoted the expression of genes related to milk fat,milk protein,and lactose synthesis in MT(P<0.05 or P<0.01).Network pharmacology predicted that the Wnt signaling pathway might be a key pathway for CRKL in treating postpartum hypogalactia.The molecular docking results showed that related chemical components in CRKL had good binding ability with CCND1 and SFRP2.Compared with the model group,CRKL at all doses inhibited the expression of SFRP2 gene in vivo(P<0.01)and activated the mRNA and protein expression of CCND1 and c-Myc in the Wnt/β-catenin signaling pathway in MT(P<0.05 or P<0.01).Cell experiments showed that,compared to the normal group,SFRP2 overexpression reduced the mRNA expression of milk synthesis-related genes FASN,CSN2,and GLUT1 in RMEC(P<0.01).The CCK8 results indicated that 10% of the drug-containing serum was the effective concentration administered to cells(P<0.01).After administering drug-containing serum,the expression of the lactation-related genes FASN,CSN2,and GLUT1 were up-regulated(compared with the SFRP2 overexpression group,P<0.01).Conclusion CRKL alleviates postpartum hypogalactia through the SFRP2-Wnt/β-catenin signaling pathway.SFRP2 might be a potential new target for the diagnosis and treatment of postpartum hypogalactia.This reveals a new mechanism of CRKL in treating postpartum hypogalactia and promotes its clinical application.