Objective: To investigate the potential protective effect of Shexiang Tongxin dropping pills (STDP) on ischemia-reperfusion injury and its underlying mechanisms in improving endothelial cell function in coronary microvascular disease (CMVD). Methods: A rat model of myocardial ischemia-reperfusion injury with CMVD was established using ligation and reperfusion of the left anterior descending artery. The effect of STDP (21.6 mg/kg) on cardiac function was evaluated using echocardiography, hematoxylin-eosin staining, and Evans blue staining. The effects of STDP on the microvascular endothelial barrier were assessed based on nitric oxide production, endothelial nitric oxide synthase expression, structural variety of tight junctions (TJs), and the expression of zonula occludens-1 (ZO-1), claudin-5, occludin, and vascular endothelial (VE)-cadherin proteins. The mechanisms of STDP (50 and 100 ng/mL) were evaluated by examining the expression of sphingosine 1-phosphate receptor 2 (S1PR2), Ras Homolog family member A (RhoA), and Rho-associated coiled-coil-containing protein kinase (ROCK) proteins and the distribution of ZO-1, VE-cadherin, and F-actin proteins in an oxygen and glucose deprivation/reoxygenation model. Results: The administration of STDP on CMVD rat model significantly improved cardiac and microvascular endothelial cell barrier functions (all P < .05). STDP enhanced the structural integrity of coronary microvascular positioning and distribution by clarifying and completing TJs and increasing the expression of ZO-1, occludin, claudin-5, and VE-cadherin in vivo (all P < .05). The S1PR2/RhoA/ROCK pathway was inhibited by STDP in vitro, leading to the regulation of endothelial cell TJs, adhesion junctions, and cytoskeletal morphology. Conclusion STDP showed protective effects on cardiac impairment and microvascular endothelial barrier injury in CMVD model rats induced by myocardial ischemia-reperfusion injury through the modulation of the S1PR2/RhoA/ROCK pathway.

Polysaccharides are the important material basis of traditional Chinese medicine（TCM）， and have various pharmacological activities such as immunomodulation， antitumor and anti-aging. Due to the large molecular weight of TCM polysaccharides， their structural analysis and oral absorption mechanism are facing technical challenges， and the current research on their structure-activity relationships has made some breakthroughs， while the research on their oral absorption mechanisms is relatively slow. In-depth study of the oral absorption mechanism of TCM polysaccharides is not only crucial for the interpretation of their action pathways and efficacy in vivo， but also helpful for the interpretation of their pharmacological effects， rational clinical applications and the discovery of new targets. In recent years， the application of fluorescent labeling and isotopic labeling methods has provided new technical means for the oral absorption studies of polysaccharides， which has promoted the development of oral absorption studies of TCM polysaccharides. In this paper， we reviewed the oral absorption pathways and labeling techniques of TCM polysaccharides， and concluded that they can be absorbed orally through transmembrane， cellular bypass， and M-cell-mediated transport， of which transmembrane pathway is the main absorption pathway， and summarized the labeling reactions of four fluorescent labeling and isotopic labeling methods with TCM polysaccharides， which can provide references for evaluating the absorption pathways of TCM polysaccharides， screening active TCM polysaccharides， establishing pharmacodynamic models and comprehensively elucidating the mechanism of TCM polysaccharides.

Objective:Super-enhancer-associated genes may be closely related to the progression of osteosarcoma,curcumin exhibits a certain inhibitory effect on tumors such as osteosarcoma.This study aims to investigate the effects of curcumin on osteosarcoma in vitro and in vivo,and to determine whether curcumin can inhibit the progression of osteosarcoma by suppressing the expression of super-enhancer-associated genes LIM and senescent cell antigen-like-containing domain 1(LIMS1),secreted protein acidic and rich in cysteine(SPARC),and sterile alpha motif domain containing 4A(SAMD4A). Methods:Human osteosarcoma cell lines(MG63 cells or U2OS cells)were treated with 5 to 50 μmol/L curcumin for 24,48,and 72 hours,followed by the methyl thiazolyl tetrazolium(MTT)assay to detect cell viability.Cells were incubated with dimethyl sulfoxide(DMSO)or curcumin(2.5,5.0 μmol/L)for 7 days,and a colony formation assay was used to measure in vitro cell proliferation.After treatment with DMSO or curcumin(10,15 μmol/L),a scratch healing assay and a transwell migration assay were performed to evaluate cell migration ability.Real-time reverse transcription polymerase chain reaction(real-time RT-PCR)and Western blotting were used to detect mRNA and protein expression levels of LIMS1,SPARC,and SAMD4A in the cells.An osteosarcoma-bearing nude mouse model was established,and curcumin was administered via gavage for 14 days to assess the impact of curcumin on tumor volume and weight in vivo.Real-time RT-PCR was used to measure mRNA expression levels of LIMS1,SPARC,and SAMD4A in the cancer and adjacent tissues from 12 osteosarcoma patients. Results:After treating cells with different concentrations of curcumin for 24,48,and 72 hours,cell viability were all significantly decreased.Compared with the DMSO group,the colony formation rates in the 2.5 μmol/L and 5.0 μmol/L curcumin groups significantly declined(both P＜0.01).The scratch healing assay showed that,compared with the DMSO group,the migration rates of cells in the 10 μmol/L and 15 μmol/L curcumin groups were significantly reduced.The exception was the 10 μmol/L curcumin group at 24 h,where the migration rate of U2OS cells did not show a statistically significant difference(P＞0.05),while all other differences were statistically significant(P＜0.01 or P＜0.001).The transwell migration assay results showed that the number of migrating cells in the 10 μmol/L and 15 μmol/L curcumin groups was significantly lower than that in the DMSO group(both P＜0.001).In the in vivo tumor-bearing mouse experiment,the curcumin group showed a reduction in tumor mass(P＜0.01)and a significant reduction in tumor volume(P＜0.001)compared with the control group.Compared with the DMSO group,the mRNA expression levels of LIMS1,SPARC,and SAMD4A in the 10 μmol/L and 15 μmol/L curcumin groups were significantly down-regulated(all P＜0.05).Additionally,the protein expression level of LIMS1 in U2OS cells in the 10 μmol/L curcumin group was significantly lower than that in the DMSO group(P＜0.05).Compared with adjacent tissues,the mRNA expression level of SPARC in osteosarcoma tissues was significantly increased(P＜0.00l),while the mRNA expression levels of LIMS1 and SAMD4A did not show statistically significant differences(both P＞0.05). Conclusion:Curcumin inhibits the proliferation and migration of osteosarcoma both in vitro and in vivo,which may be associated with the inactivation of super-enhancer-associated gene LIMS1.

Objective To explore the antimicrobial resistance of carbapenem-resistant Klebsiella pneumoniae(CRKP)isolated from blood and the related risk factors for infection in patients.Methods Clinical data of 383 KP-infected patients from whose blood Klebsiella pneumoniae(KP)were isolated during hospitalization period in a hos-pital from January 2018 to December 2021 were retrospectively analyzed.Patients were divided into CRKP group(n=114)and non-CRKP group(n=269)based on antimicrobial resistance.According to the prognosis,114 patients in the CRKP group were subdivided into the death group(n=30)and the survival group(n=84).General informa-tion,underlying diseases,antimicrobial use,and infection outcomes of two groups of patients were compared,and risk factors for infection and death after infection were analyzed.Results The resistance rates of KP to tigecycline and compound sulfamethoxazole showed upward trends,with statistically significant differences(both P=0.008).The CRKP group had higher resistance rates to amikacin,aztreonam,compound sulfamethoxazole,ciprofloxacin,cefepime,cefoperazone/sulbactam,piperacillin/tazobactam,tigecycline,ceftazidime,tobramycin,and levofloxacin,as well as higher in-hospital mortality than the non-CRKP group,with statistically significant differences(all P＜0.05).Acute pancreatitis prior to infection(OR=16.564,P＜0.001),hypoalbuminemia(OR=8.588,P＜0.001),stay in in-tensive care unit prior to infection(OR=2.733,P=0.017),blood transfusion(OR=3.968,P=0.001),broncho-scopy(OR=5.194,P=0.014),surgery within 30 days prior to infection(OR=2.603,P=0.010),and treatment with carbapenems(OR=2.663,P=0.011)were independent risk factors for the development of CRKP blood-stream infection(BSI).Cardiac insufficiency before infection(OR=11.094,P=0.001),combined with pulmonary infection(OR=20.801,P=0.010),septic shock(OR=9.783,P=0.002),disturbance of consciousness(OR=11.648,P=0.001),and receiving glucocorticoid treatment(OR=5.333,P=0.018)were independent risk factors for mortality in patients with CRKP BSI.Conclusion The resistance rate of KP from BSI to tigecycline and com-pound sulfamethoxazole presents upward trend.Underlying diseases,invasive procedures,and carbapenem treat-ment are closely related to CRKP BSI.Cardiac insufficiency,pulmonary infection,septic shock,disturbance of con-sciousness,and glucocorticoid treatment can lead to death of patients with CRKP BSI.

The Baimai Ointment with the effect of relaxing sinew and activating collaterals demonstrates a definite effect on Baimai disease with pain, spasm, stiffness and other symptoms, while the pharmacodynamic characteristics and mechanism of this agent remain unclear. In this study, a rat model of chronic compression of L4 dorsal root ganglion(CCD) was established by lumbar disc herniation, and the efficacy and mechanism of Baimai Ointment in the treatment of CCD were preliminarily explored by behavioral tests, side effect evaluation, network analysis, antagonist and molecular biology verification. The pharmacodynamic experiment indicated that Baimai Ointment significantly improved the pain thresholds(mechanical pain, thermal pain, and cold pain) and gait behavior of CCD model rats without causing tolerance or obvious toxic and side effects. Baimai Ointment inhibited the second-phase nociceptive response of mice in the formalin test, increased the hot plate threshold of normal mice, and down-regulated the expression of inflammatory cytokines in the spinal cord. Network analysis showed that Baimai Ointment had synergistic effect in the treatment of CCD and was related to descending inhibition/facilitation system and neuroinflammation. Furthermore, behavioral tests, Western blot, and immunofluorescence assay revealed that the pain-relieving effect of Baimai Ointment on CCD may be related to the regulation of the interaction between neuroactive ligand and receptors(neuroligands) such as CHRNA7, ADRA2A, and ADRB2, and the down-regulation of the expression of NOS2/pERK/PI3K, the core regulatory element of HIF-1 signaling pathway in spinal microglia. The findings preliminarily reveal the mechanism of relaxing sinew and activating collaterals of Baimai Ointment in the treatment of Baimai disease, providing a reference for the rational drug use and further research of this agent.
Rats ; Mice ; Animals ; Chronic Pain/metabolism* ; Rats, Sprague-Dawley ; Ganglia, Spinal/metabolism* ; Ligands ; Signal Transduction ; Hyperalgesia/metabolism* ; Drugs, Chinese Herbal

Lignans derived from Eucommia ulmoides Oliver (Eucommia lignans) inhibit the progression of inflammatory diseases, while their effect on the progression of diabetic nephropathy (DN) remained unclear. This work was designed to assess the function of Eucommia lignans in DN. The major constituents of Eucommia lignans were analyzed by UPLC-Q-TOF-MS/MS. The binding between Eucommia lignans and aldose reductase (AR) was predicted by molecular docking. Eucommia lignans (200, 100, and 50 mg·kg^-1) were used in model animals to evaluate their renal function changes. Rat glomerular mesangial cells (HBZY-1) were transfected with sh-AR, sh-AMPK, and oe-AR in the presence of high glucose (HG) or HG combined with Eucommia lignans to evaluate whether Eucommia lignans affected HG-induced cell injury and mitochondrial dysfunction through the AR/Nrf2/HO-1/AMPK axis. Eucommia lignans significantly attenuated the progression of DN in vivo. Eucommia lignans notably reversed HG-induced upregulation of inflammatory cytokines and mitochondrial injury, while downregulating the levels of Cyto c, caspase 9, AR, and NOX4 in HBZY-1 cells. In contrast, HG-induced downregulation of Nrf2, HO-1 and p-AMPKα levels were abolished by Eucommia lignans. Meanwhile, knockdown of AR exerted similar therapeutic effect of Eucommia lignans on DN progression, and AR overexpression reversed the effect of Eucommia lignans. Eucommia lignans alleviated renal injury through the AR/Nrf2/HO-1/AMPK axis. Thus, these findings might provide evidence for the use of Eucommia lignans in treating DN.
Animals ; Rats ; AMP-Activated Protein Kinases/genetics* ; Diabetes Mellitus ; Diabetic Nephropathies/prevention & control* ; Eucommiaceae/metabolism* ; Lignans/therapeutic use* ; Molecular Docking Simulation ; NF-E2-Related Factor 2/metabolism* ; Tandem Mass Spectrometry

ObjectiveA rapid method for identification of chemical constituents in Puerariae Lobatae Radix dispensing granules was established in order to clarify the material basis. MethodThe chemical constituents of Puerariae Lobatae Radix dispensing granules was qualitatively analyzed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） under positive and negative ion modes， and the chromatographic conditions were on an ACQUITY UPLC HSS T3 column（2.1 mm×100 mm， 1.8 μm） with 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） as mobile phase for gradient elution（0-4 min， 5%-10%B； 4-10 min， 10%-15%B； 10-20 min， 15%-16%B； 20-27 min， 16%-31%B； 27-33 min， 31%-59%B； 33-42 min， 59%-95%B； 42-42.1 min， 95%-5%B； 42.1-45 min， 5%B）， the flow rate was 0.35 mL·min^-1， the column temperature was 40 ℃， the injection volume was 5 μL， and electrospray ionization（ESI） was selected. Then these chemical constituents were comprehensively identified based on PeakView 1.2， PubChem， ChemicalBook， ChemSpider， comparative control profiles and literature information. ResultA total of 128 chemical constituents were identified from the dispensing granules， including 60 flavonoids， 26 organic acids， 7 glycosides， 6 coumarins， 3 nucleosides and 26 other compounds. By focusing on the cleavage patterns of flavonoids， organic acids， glycosides， coumarins， nucleosides and other compounds， 12 compounds that have not been reported in Puerariae Lobatae Radix species were identified from the dispensing granules. ConclusionThe established method can systematically and rapidly identify the chemical constituents in Puerariae Lobatae Radix dispensing granules， and cleared it composition is mainly flavonoids and organic acids. Laying a foundation for the study of the material basis， mechanism of action and clinical application of the dispensing granules.

Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. dujiao-longae. The phylogenetic tree could distinguish them effectively. HPLC fingerprints of 18 batches of C. cicadae and 5 batches of T. dujiaolongae were established. The results of CA, PCA, and PLS-DA were consistent, which could distinguish them well, indicating that there were great differences in chemical components between C. cicadae and T. dujiaolongae. The results of PLS-DA showed that six components such as uridine, guanosine, adenosine, and N~6-(2-hydroxyethyl) adenosine were the main differential markers of the two species. ITS sequences and HPLC fingerprint combined with the chemical pattern recognition method can serve as the identification and differentiation methods for C. cicadae and T. dujiaolongae.
Chromatography, High Pressure Liquid/methods* ; Cordyceps/genetics* ; Hypocreales ; Phylogeny

Objective:To explore the risk factors and prognosis of carbapenem-resistant Klebsiella pneumoniae (CRKP) infection for inpatients in hepatobiliary surgery. Methods:The clinical data of patients with Klebsiella pneumoniae infection admitted to the Department of Hepatobiliary Surgery, Hunan Provincial People's Hospital from January 2016 to December 2020 were retrospectively analyzed. For each patient with CRKP infection, two patients with non-carbapenem-resistant Klebsiella pneumoniae (non-CRKP) infection were selected for matching. A total of 720 patients with Klebsiella pneumoniae infection were involved, including 444 males and 276 females, aged (58.0±11.6) years old. According to the infection conditions, they were divided into two groups: CRKP group ( n=240) and non-CRKP group ( n=480). The 240 CRKP patients were divided into two subgroups according to their prognosis: death group ( n=34) and survival group ( n=206). The general information, laboratory test results, antibiotic use and infection outcomes of patients were recorded to analyze the risk factors of infection and death after infection. Results:Acute pancreatitis ( OR=3.473, 95% CI: 1.844-6.541), chronic cardiovascular disease before infection ( OR=2.028, 95% CI: 1.228-3.347), chronic renal failure ( OR=1.873, 95% CI: 1.142-3.073), hypoalbuminemia ( OR=3.060, 95% CI: 1.869-5.010), use of carbapenems ( OR=3.882, 95% CI: 2.518-5.985), admission to intensive care unit ( OR=1.783, 95% CI: 1.034-3.075) and surgery within 30 days before infection ( OR=13.463, 95% CI: 7.482-24.223) were independent risk factors for CRKP infection inpatients in hepatobiliary surgery(all P<0.05). Chronic respiratory disease before infection ( OR=3.784, 95% CI: 1.420-10.089), mechanical ventilation ( OR=5.085, 95% CI: 1.436-18.011), disturbance of consciousness ( OR=40.710, 95% CI: 3.564-464.943), hormone therapy ( OR=14.977, 95% CI: 3.819-58.743) and treatment of quinolone antibiotics ( OR=4.102, 95% CI: 1.226-13.726) were independent risk factors for death of inpatients with CRKP infection in hepatobiliary surgery (all P<0.05). The resistance of amikacin, tobramycin, ceftazidime, cefepime, aztreonam, ciprofloxacin, levofloxacin, co-sulfamethoxazole and piperacillin/tazobactamand in CRKP group were significantly different compared with non-CRKP group (all P<0.05). Conclusion:The occurrence of CRKP infection for inpatients in hepatobiliary surgery is related to various factors such as underlying diseases, antibiotic use and self-barrier destruction, and these factors affect the infection outcome of patients.

Objective: To observe the clinical efficacy of acupuncture and moxibustion for functional dyspepsia (FD) due to Yang deficiency of the spleen and stomach.Methods: A total of 90 patients meeting the inclusion criteria of FD due to Yang deficiency of the spleen and stomach were randomized into an acupuncture-moxibustion group, an electroacupuncture (EA) group, and a mosapride group by the random number table method, with 30 cases in each group. The mosapride group was treated with mosapride citrate dispersible tablets. The EA group was treated with EA treatment. The acupuncture-moxibustion group was treated with additional moxibustion therapy based on the treatment in the EA group. All three groups took 10 d as one treatment course and 2 courses in total, with a 2-day interval between two courses. The traditional Chinese medicine (TCM) symptoms score, functional digestive disorder quality of life questionnaire (FDDQL) score, oral-colon transit time (OCTT), and serum glucagon-like peptide-1 (GLP-1) level was compared before and after treatment among the three groups. And the clinical efficacy was evaluated.Results: The total effective rate in the acupuncture-moxibustion group was 96.7％, which was higher than 86.7％ in the EA group and 73.3％ in the mosapride group, and the total effective rate in the EA group was higher than that in the mosapride group; the inter-group differences were all statistically significant (P<0.05). After treatment, the TCM symptom score in the three groups was lower than that before treatment (P<0.05), and the FDDQL score was higher than that before treatment (P<0.05). The change after treatment in the TCM symptom score in the acupuncture- moxibustion group and the EA group was more significant than that in the mosapride group, and the change in the acupuncture-moxibustion group was more significant than that in the EA group; the inter-group differences were statistically significant (P<0.05). The change after treatment in the FDDQL score in the acupuncture-moxibustion group and the EA group was greater than that in the mosapride group, and the change in the acupuncture-moxibustion group was larger than that in the EA group; the inter-group differences were all statistically significant (P<0.05). After treatment, the OCTT in all three groups was lower than that before treatment (P<0.05), and the serum GLP-1 level was higher than that before treatment (P<0.05); the change after treatment in the OCTT in the acupuncture-moxibustion group and the EA group was more significant than that in the mosapride group, and the change in the acupuncture- moxibustion group was more significant than that in the EA group; the inter-group differences were statistically significant (P<0.05). The change after treatment in the serum GLP-1 level in the acupuncture-moxibustion group and the EA group was greater than that in the mosapride group, and the change in the acupuncture-moxibustion group was larger than that in the EA group; the inter-group differences were statistically significant (P<0.05).Conclusion: Acupuncture and moxibustion can improve the clinical symptoms in patients with FD and enhance their quality of life, producing better effects than EA or mosapride alone. Shortening gastrointestinal movement time and increasing gastrointestinal hormone levels may be its mechanism.