The differences in chemical quality characteristics between Xinjiang Rehmannia glutinosa and R. glutinosa were analyzed to provide a theoretical basis for the introduction and quality control of R. glutinosa. In this study, the high performance liquid chromatography(HPLC) fingerprints of 6 batches of Xinjiang R. glutinosa and 10 batches of R. glutinosa samples were established. The content of iridoid glycosides, phenylethanoid glycosides, monosaccharides, oligosaccharides, and polysaccharides in Xinjiang R. glutinosa and R. glutinosa was determined by high performance liquid chromatography-diode array detection(HPLC-DAD), high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD), and ultraviolet-visible spectroscopy(UV-Vis). The determination results were analyzed with by chemical pattern recognition and entropy weight TOPSIS method. The results showed that there were 19 common peaks in the HPLC fingerprints of the 16 batches of R. glutinosa, and catalpol, aucubin, rehmannioside D, rehmannioside A, hydroxytyrosol, leonuride, salidroside, cistanoside A, and verbascoside were identified. Hierarchical cluster analysis(HCA) and principal component analysis(PCA) showed that Qinyang R. glutinosa, Mengzhou R. glutinosa, and Xinjiang R. glutinosa were grouped into three different categories, and eight common components causing the chemical quality difference between Xinjiang R. glutinosa and R. glutinosa in Mengzhou and Qinyang of Henan province were screened out by orthogonal partial least squares discriminant analysis(OPLS-DA). The results of content determination showed that there were glucose, sucrose, raffinose, stachyose, polysaccharides, and nine glycosides in Xinjiang R. glutinosa and R. glutinosa samples, and the content of catalpol, rehmannioside A, leonuride, cistanoside A, verbascoside, sucrose, and glucose was significantly different between Xinjiang R. glutinosa and R. glutinosa. The analysis with entropy weight TOPSIS method showed that the comprehensive quality of R. glutinosa in Mengzhou and Qinyang of Henan province was better than that of Xinjiang R. glutinosa. In conclusion, the types of main chemical components of R. glutinosa and Xinjiang R. glutinosa were the same, but their content was different. The chemical quality of R. glutinosa was better than Xinjiang R. glutinosa, and other components in R. glutinosa from two producing areas and their effects need further study.
Rehmannia/classification* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Quality Control

BACKGROUND:Most rotator cuff injuries occur in the supraspinatus tendon.Clinical treatment of rotator cuff injuries is very limited due to the lack of blood vessels and the complex anatomical structure of the rotator cuff.The rapid development of tissue engineering technology and stem cell biology has brought new hope for improving the quality of tendon repair.OBJECTIVE:To prepare human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffolds by bio-3D printing technology to observe the effect of this scaffold on repairing rotator cuff injury.METHODS:(1)In vitro cell assay:The gelatin microcarrier was prepared.The tissue engineered stem cells were constructed by inoculating human umbilical cord mesenchymal stem cells on the surface of gelatin microcarrier.Gelatin methacrylate hydrogel printing ink was prepared.Tissue engineered stem cells were re-suspended with gelatin methacrylate hydrogel printing ink and put into the bio-ink container of 3D printer for printing.Human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffold was obtained after 5 minutes of blue light irradiation and curing.The activity of human umbilical cord mesenchymal stem cells in scaffolds was detected by dead/alive staining and CCK-8 assay.(2)In vivo animal experiments:A random block design method was used to randomly assign 24 SD rats to 4 groups with 6 rats in each group.No treatment was given in the normal group.The rotator cuff injury model of supratinatus tendon tear was established in the rotator cuff injury group,the simple scaffold group,and the cellular scaffold group.The gelatin methacrylate scaffold and human umbilical cord mesenchymal stem cell/gelatin methacrylatecomposite scaffold were implanted into the tendon injury after the model was made in the simple scaffold group and the cellular scaffold group,respectively.Four weeks after operation,behavioral tests and histopathological morphology observation of supraspinatus tendon of rotator cuff were performed.RESULTS AND CONCLUSION:(1)In vitro cellular assay:The dead/alive staining showed that gelatin microcarrier could reduce the damage of human umbilical cord mesenchymal stem cells caused by 3D printing process.With the extension of culture time,the survival rate of human umbilical cord mesenchymal stem cells increased in the scaffold.The results of CCK-8 assay showed that with the extension of culture time,the activity of human umbilical cord mesenchymal stem cells in the scaffold did not change significantly.(2)In vivo animal experiments:Behavioral test results showed that compared with rotator cuff injury group and simple scaffold group,cellular scaffold group significantly improved limb motor function.The results of hematoxylin-eosin and Masson staining of rotator cuff supraspinatus tendon showed that compared with rotator cuff injury group and simple scaffold group,the muscle fiber arrangement in the cellular scaffold group was more regular;there was no obvious inflammatory cell infiltration,and the percentage of collagen volume decreased.The results of immunofluorescence staining showed that the expression levels of interleukin 6 and tumor necrosis factor α in the rotator cuff supraspinatus tendon were significantly decreased in the cellular scaffold group compared with the rotator cuff injury group and the simple scaffold group.(3)The results showed that bio-3D-printed cell scaffolds encapsulating human umbilical cord mesenchymal stem cells/gelatin methacrylate could promote tissue repair and regeneration of rotator cuff injuries.

Objective: To explore the effects of different types of acute exercise on the working memory of sedentary college students，so as to provide a basis for exercise intervention. Methods: From April 15 to May 30, 2023, a total of 42 sedentary college students were recruited from one university in Beijing. Using a single blind, completely randomized experimental design, participants were randomly assigned to an open skill exercise group, a closed skill exercise group, or a control group, with 14 participants in each group. The open skill exercise group engaged in 30 minutes of badminton, the closed skill exercise group performed 30 minutes of running, and the control group remained seated for 30 minutes. All participants completed a 2-back working memory task and had their electroencephalogram (EEG) data recorded before and after the intervention. Results: The accuracy rates of the open skill exercise group, closed skill exercise group, and control group (0.90±0.06, 0.94±0.05; 0.88±0.05, 0.94±0.05; 0.85±0.10, 0.90±0.06) showed a significant main effect of time ( F=37.14, P <0.01). Reaction times [(923.65±145.08, 711.56± 140.93 ; 909.59±180.28, 807.85±169.66; 917.05±166.35, 871.86±186.07)ms] showed both a significant main effect of time and a significant interaction between group and time ( F=70.55, 11.83, P <0.01). Repeated measures ANOVA revealed that all three groups improved in accuracy and reaction time compared to pre test values, with no significant difference in accuracy between groups. However, the reaction time of the open skill exercise group was significantly faster than that of the control group ( P <0.05), while there was no significant difference between the closed skill exercise group and the control group ( P >0.05). For EEG data, the P2 amplitude showed a significant main effect of time and a significant interaction between groups and time ( F=10.60, 7.66, P < 0.01 ), with the open skill exercise group exhibiting a higher P2 amplitude than the control group ( P <0.05), while the closed skill exercise group showed no significant difference compared to the control group ( P >0.05). The N2 amplitude showed a significant main effect of time ( F=5.94, P <0.05). The P3 amplitude showed significant main effects of time and electrode position, as well as a significant interaction between groups and time ( F=23.16, 4.53, 5.85, P <0.05), with both exercise groups exhibiting higher P3 amplitudes than the control group ( P <0.05), but no significant difference between the two exercise groups ( P >0.05). Conclusion Open skill exercise is more effective than closed skill exercise in improving the working memory of sedentary college students.

BACKGROUND:The absence of blood vessels in tendon tissue makes tendon repair challenging.Therefore,improving tendon healing and raising the efficacy of stem cell and other therapeutic cell transplantation after tendon damage have become hotspots for research in both clinical and scientific contexts. OBJECTIVE:The stem cells and gelatin microcarrier scaffold were joined to form tissue engineered stem cells.Human umbilical cord mesenchymal stem cells cultured in gelatin microcarriers were used to investigate the therapeutic impact and mode of action on tendinopathy healing in rats in vitro and In vivo. METHODS:(1)In vitro cell experiments:After seeding human umbilical cord mesenchymal stem cells with three-dimensional gelatin microcarriers,the cell vitality and survival were assessed.Human umbilical cord mesenchymal stem cells conventionally cultured were cultured as controls.(2)In vivo experiment:Adult SD rats were randomly assigned to normal group,tendinopathy group,2D group(tendinopathy+conventional culture of human umbilical cord mesenchymal stem cells),and 3D group(tendinopathy+gelatin microcarrier three-dimensional culture of human umbilical cord mesenchymal stem cells),with 6 rats in each group.Four weeks after therapy,animal behavior tests and histopathologic morphology of the Achilles tendon was examined. RESULTS AND CONCLUSION:(1)In vitro cell experiments:the seeded human umbilical cord mesenchymal stem cells on gelatin microcarriers showed high viability and as time went on,the stem cell proliferation level grew.Compared with the control group,3D stem cell culture preserved cell viability.(2)In vivo experiment:Following a 4-week treatment,the 3D stem cell culture group showed a significant improvement in both functional recovery of the lower limbs and histopathological scores when compared to the tendinopathy group.The 2D stem cell culture group also showed improvement in tendinopathy injury,but its effect is not as much as the 3D stem cell culture group.(3)The outcomes demonstrate that human umbilical cord mesenchymal stem cells cultured with three-dimensional gelatin microcarrier can promote the repair and regeneration of tendon injury tissue,and the repair effect is better than that of conventional human umbilical cord mesenchymal stem cells.

Objective To establish a stable and reproducible animal model of abdominal intestinal firearm penetrating injury in a cold high-altitude environment.Methods Twenty landrace pigs were randomly and equally assigned to a low-altitude normal temperature(LN)group and a high-altitude cold(HC)group.The HC group was placed in a cold environment at high altitudes,and the LN group was placed in a normal-temperature environment at low altitudes.They were raised for 48 hours respectively.After anesthesia,they were suspended on the shooting range,and the right lower abdomen of the experimental pigs was shot with a gun.After injury,they were simply bandaged and transported back to the laboratory for observation in the normal temperature environment of the low altitudes.The vital signs and injuries at 0,2,4,8,12 and 24 h and 24 h survival rates of experimental pigs were compared.Laparotomy was immediately performed on the dead pigs and the experimental pigs still alive at 24 h to explore the injuries and observe the pathology of the small intestine and colon.Results The 24 h survival rate of the HC group was 70％,with no statistically significant difference compared to the LN group's 90％(P＞0.05).After the injury,the body temperature of both groups gradually increased.The body temperature of the HC group was significantly higher than the LN group at 0,2,4 and 8 h time points(P＜0.001),and the LN group exceeded the HC group at 24 h(P＜0.05).Both groups showed an initial increase followed by a decrease in heart rate,with the HC group significantly higher than the LN group only at 0 h(P＜0.01),and no statistically significant differences were observed at other time points(P＞0.05).Both groups showed an early increase and later decrease in respiratory rate,with the HC group higher than the LN group at 0,4,8,12 and 24 h(P＜0.05 or P＜0.001).There was no statistically significant difference(P＞0.05)between the HC group and the LN group in small intestine rupture,small intestine contusion,mesenteric injury,colon rupture and wound diameter.The pathology of the small intestine and colon in the HC group showed extensive necrosis and shedding of the mucosa layer,severe congestion and edema of the submucosa,and extensive lymphocyte infiltration.The LN group also showed similar symptoms but to a lesser extent.Conclusion This study established a pig model of abdominal firearm intestinal perforation injury in a cold environment at high-altitudes.The model has strong operability and stable damage,which can provide a reference for subsequent research.

With the increasing demand for dynamic,real-time and rapid qualitative analysis of chemical composition in areas such as emergency response and space exploration,chip-scale mass spectrometers have attracted significant attention.These devices are expected to drive the integration of mass spectrometry with micro/nano-fabrication and intelligent sensing technologies,fostering profound innovation and breakthroughs in analytical chemistry.As an excellent mass analyzer,the ion trap exhibits numerous advantages,and its miniaturization creates favorable conditions for the high-density integration of miniature mass spectrometers.However,the reduction in ion storage capacity may compromise its sensitivity and dynamic range,rendering the study of ion unidirectional ejection in highly miniaturized ion traps of significant practical importance.In this work,a research was conducted on achieving efficient ion unidirectional ejection while maintaining high mass resolution in the extremely miniature hyperbolic linear ion trap(M-HLIT)with a field radius of 1 mm,and an electric field compensation method was proposed,which combined asymmetric electrode stretching and unbalanced RF voltage to achieve high-precision optimization of the electric field composition.Simulations showed that in an ideal structure,this method achieved 100％unidirectional ejection efficiency with the mass resolution of 518,significantly outperforming traditional asymmetric structure method(365)and unbalanced voltage method(321).Following the introduction of ion ejection slots,further optimization through bidirectional stretching and electrical parameters improved the resolution to 790 while maintaining a unidirectional ejection efficiency of 93％.This method eliminated the requirement for additional excitation voltage,offering an ideal solution for the miniature mass analyzer with high detection performance of chip-level mass spectrometers.

Objective To verify the efficacy of a domestic human DNA quantification kit based on real-time fluorescence quantitative PCR in detecting the total human DNA concentration,male DNA concen-tration in mixed male/female DNA samples,the degree of DNA degradation and inhibitor tolerance.Methods Samples with different concentrations,different male/female ratios,different concentrations of inhibitors,and different degradation degrees were tested using the domestic human DNA quantification kit based on real-time fluorescence quantitative PCR.This kit was compared with a similar product on the market and was applied to the detection of DNA from real cases.Results This human DNA quan-tification kit can effectively detect human DNA as low as 0.001 65 ng/μL,and 6.25 pg/μL of male DNA in mixed samples with a male-to-female ratio of 1∶15 000.Even when the sample contains as high as 400 ng/μL of humic acid or 1 000 μmol/L of hemin alone,the DNA concentration can still be accurately detected.The degradation index can effectively characterize the degradation degree of the sample.This kit has been successfully applied in forensic practice.Conclusion This human DNA quan-tification kit is accurate and reliable in detection.It can accurately reflect the degradation of DNA and inhibitor tolerance.It has good performance in quantitative accuracy,determination of the male/female ratio in mixed samples,and inhibitor tolerance.It has application potential in forensic case examination.

Objective To analyze the effect of sodium cantharidinate and vitamin B6 injection(SCV)on four human hepatocellular carcinoma(HCC)cell lines(SMMC-7721,Bel-7402,Huh7,and HepG2)and explore its mechanism.Methods Normal hepatic cell line L02 was treated with SCV at concentrations of 0 μmol/L(control),0.5,1,2,4,8,16,and 32 μmol/L,and the cytotoxicity of SCV on L02 cells was detected using CCK-8 assay.Human HCC cell lines(SMMC-7721,Bel-7402,Huh7,and HepG2)were cultured.SCV-untreated control group(0 μmol/L)and 2,4,and 8 μmol/L SCV-treated groups were set up.CCK-8 assay,plate cloning formation assay,Transwell assay,wound healing assay,and flow cytometry were used to detect the effects of SCV on the growth and proliferation capacity,colony formation ability,invasion and migration capabilities,cell cycle,and apoptosis of the four hepatocellular carcinoma cell lines,respectively.Western blotting was performed to detect the expression levels of apoptosis-related proteins,including nuclear factor kappa-B subunit p65(p65),B-cell lymphoma 2(Bcl-2),and Caspase-3,and to preliminarily explore the underlying mechanism.Results The CCK-8 assay showed that SCV at 0.5,1,2,4,and 8 μmol/L had no significant cytotoxic effect on L02 cells compared with untreated control group,so 2,4,and 8 μmol/L SCV were selected for subsequent experiments.Compared with the untreated control group(0 μmol/L),SCV at different concentrations(2,4,and 8 μmol/L)significantly inhibited the proliferation of the four HCC cell lines(P<0.001).The plate cloning formation assay showed that SCV at different concentrations(2,4,and 8 μmol/L)significantly reduced the colony formation ability of the four HCC cell lines(P<0.05 or P<0.01 or P<0.001).In addition,Transwell and wound healing assays revealed that SCV at different concentrations(2,4,and 8 μmol/L)significantly inhibited the invasion and migration of HCC cells(P<0.05 or P<0.01 or P<0.001).In the above results,the inhibitory effect of SCV was concentration-dependent.Flow cytometry analysis indicated that SCV arrested cells in the G2/M phase(P<0.05 or P<0.01 or P<0.001)and significantly promoted cell apoptosis(P<0.05 or P<0.01 or P<0.001).Western blotting showed that SCV significantly down-regulated the expression of p65(P<0.05 or P<0.01)and Bcl-2(P<0.05),and up-regulated the expression of Caspase-3(P<0.05 or P<0.01).Conclusions SCV can significantly inhibit the proliferation,colony formation,invasion,and migration of multiple human HCC cell lines and arrest the cell cycle.SCV may inhibit the expression of p65 and Bcl-2,thereby lifting their inhibitory effect on the apoptotic pathway and activating Caspase-3 to promote apoptosis.

Objective:To discuss the differential expression of microRNA(miR)-125a-5p in peripheral blood mononuclear cells(PBMCs)of the patients with mycobacterium tuberculosis(MTB)infection and its effect on macrophage polarization,and to clarify its clinical significance.Methods:A total of 40 patients with active tuberculosis(ATB)(ATB group),35 patients with latent tuberculosis infection(LTBI)(LTBI group),and 40 healthy physical examinees(control group)clinically diagnosed from July 2022 to June 2023 were selected.The fasting blood samples of the subjects in three groups were collected next morning after 12 h of fasting,and then serum was separated.Enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of inflammatory factors in the serum of the subjects in various groups.Simultaneously,the PBMCs were extracted from the subjects in various groups;flow cytometry was used to detect the expression levels of CD80 and CD206 proteins in the PBMCs of the subjects in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of microRNA(miR)-125a-5p and interleukin-6(IL-6)mRNA in PBMCs of the subjects in various groups.Results:There were no statistically significant differences in the general information of the subjects among three groups(P>0.05).Compared with control group,the erythrocyte sedimentation rate(ESR),percentages of monocytes(MONO),tumor necrosis factor-α(TNF-α)levels,and interleukin-10(IL-10)levels in serum of the patients in ATB group and LTBI group were significantly increased(P<0.05),and the percentages of lymphocytes(LY)were significantly decreased(P<0.05);compared with ATB group,the ESR and level of IL-10 in serum of the patients in LTBI group were significantly decreased(P<0.05),and the percentage of LY was significantly increased(P<0.05);there were statistically significant differences in the counts of white blood cell(WBC)of the subjects among various groups(P>0.05).The flow cytometry results showed that compared with control group,the expression levels of CD80 and CD206 proteins in PBMCs of the patients in ATB group and LTBI group were significantly increased(P<0.05).Compared with ATB group,the expression level of CD206 protein in the PBMCs of the patients in LTBI group was significantly increased(P<0.05),and the expression level of CD80 protein was significantly decreased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of miR-125a-5p and IL-6 mRNA in the PBMCs of the patients in ATB group and LTBI group were significantly increased(P<0.05);compared with ATB group,the expression levels of miR-125a-5p and IL-6 mRNA in PBMCs of the patients in LTBI group were significantly increased(P<0.05).The correlation analysis results showed that the miR-125a-5p expression level was positively correlated with the TNF-α level and IL-6 mRNA expression level(r=0.406,P<0.05;r=0.351,P<0.05),and negatively correlated with the IL-10 level(r=-0.368,P<0.05).The area under the receiver operating characteristic(ROC)curve(AUC)value of miR-125a-5p expression level for diagnosing LTBI patients was 0.89(P<0.01),with a sensitivity of 0.85 and a specificity of 0.88.Conclusion:The expression level of miR-125a-5p in PBMCs of the patients in LTBI group is significantly increased,and it can affect the macrophage polarization to M1,promote the inflammatory response process of macrophages and participate in the occurrence and development of pulmonary tuberculosis.

Pyroptosis is an identified programmed cell death that has been highly linked to endoplasmic reticulum (ER) dynamics. However, the crucial proteins for modulating dynamic ER membrane curvature change that trigger pyroptosis are currently not well understood. In this study, a biotin-labeled chemical probe of potent pyroptosis inducer α-mangostin (α-MG) was synthesized. Through protein microarray analysis, reticulon-4 (RTN4/Nogo), a crucial regulator of ER membrane curvature, was identified as a target of α-MG. We observed that chemically induced proteasome degradation of RTN4 by α-MG through recruiting E3 ligase UBR5 significantly enhances the pyroptosis phenotype in cancer cells. Interestingly, the downregulation of RTN4 expression significantly facilitated a dynamic remodeling of ER membrane curvature through a transition from tubules to sheets, consequently leading to rapid fusion of the ER with the cell plasma membrane. In particular, the ER-to-plasma membrane fusion process is supported by the observed translocation of several crucial ER markers to the "bubble" structures of pyroptotic cells. Furthermore, α-MG-induced RTN4 knockdown leads to pyruvate kinase M2 (PKM2)-dependent conventional caspase-3/gasdermin E (GSDME) cleavages for pyroptosis progression. In vivo, we observed that chemical or genetic RTN4 knockdown significantly inhibited cancer cells growth, which further exhibited an antitumor immune response with anti-programmed death-1 (anti-PD-1). In translational research, RTN4 high expression was closely correlated with the tumor metastasis and death of patients. Taken together, RTN4 plays a fundamental role in inducing pyroptosis through the modulation of ER membrane curvature remodeling, thus representing a prospective druggable target for anticancer immunotherapy.
Pyroptosis/immunology* ; Humans ; Endoplasmic Reticulum/immunology* ; Animals ; Nogo Proteins/antagonists & inhibitors* ; Mice ; Cell Line, Tumor ; Xanthones/pharmacology* ; Neoplasms/pathology* ; Mice, Nude