To investigate the effects of isovitexin Ⅳ on transient outward potassium current in rat ventricular myocytes. In this study, MTT assay was used to investigate the safe range of isovitexin. The results showed that the IC₅₀ of the drug was in the range of 10-30 μmol•L⁻¹, and the drug concentration of 1-3 μmol•L⁻¹ for the patch clamp test was within the safe range. In addition, the single ventricular myocytes were obtained by single-enzymatic hydrolysis through aortic retrograde perfusion. The transient outward potassium current (Ito) of rat ventricular myocytes was guided and measured by whole-cell patch-clamp technique and the changes of current characteristics were recorded after isovite was applied. When the concentration of IV was less than 0.1 μmol•L⁻¹, there was no significant effect on Ito. However, with the increase in the concentration of IV (≥0.3 μmol•L⁻¹), the peak of Ito was decreased gradually, from (32.32±2.9) pA/pF to （25.83±4.3） pA/pF, 1 μmol•L⁻¹ IV and (19.51±3.5) pA/pF, 3 μmol•L⁻¹ IV respectively, with an inhibition effect in a concentration-dependent manner. In the range of 1-3 μmol•L⁻¹, IV down-regulated the I-V curve of Ito significantly. The activation curve showed that IV can enable the maximum half activation potential (V1/2) to move to the positive direction, and the V1/2 was increased from (19.59±1.6) mV to (22.81±1.7) mV and (28.86±1.4) mV at concentration of 1, 3 μmol•L⁻¹, meanwhile the activation curve moved to the right. However, the maximum half inactivating potential (V1/2) of the steady-state inactivation curve of Ito was significantly decreased from (－51.43±0.99) mV to (－61.81±1.3) mV with concentration of 1 μmol•L⁻¹ and (－71.50±1.4) mV with concentration of 3 μmol•L⁻¹. The inactivation time constant of recovery from inactivation (τ) was up-regulated significantly from (94.89±0.73) ms to (118.5±1.5) ms and (162.4±1.4) ms at concentration of 1, 3 μmol•L⁻¹ respectively. Meanwhile IV could enable the inactivation recovery curve to move to the right, which suggested that it can prolong the recovery time from inactivation of the transient outward potassium channel. In conclusion, isovitexin had a high inhibitory effect on Ito in rat ventricular myocytes.

OBJECTIVETo study the effect of Shuangling Fuzheng anti-tumor preparation (SLAP) five groups on proliferation and c-myc gene expression of SGC-7901 cells in vitro.

METHODThe inhibitory effect of single SLAP (40 -640 microg x mL(-1)) and combined therapy with adriamycin (0.4, 4.0 microg x mL(-1) or cisplatin (0.1,1.0 microg x mL(-1) on human gastric carcinoma SGC-7901 cells proliferation were observed by MTT colorimetric analysis method. Technique of flow cytometry in vitro was used to measure the rate of positive sign of SLAP (80 - 320 microg x mL(-1)) on c-myc gene protein of SGC-7901 cells.

RESULTSGC-7901 cells proliferation were inhibited by single SLAP in dose of 40 - 640 microg x mL(-1) 24 h. Its inhibitory rate was increased with increase of dose. The inhibitory rate on SGC-7901 cells could be increased by SLAP in dose of 40 - 640 microg x mL(-1) plus adriamycin in dose of 0.4 and 4.0 microg x mL(-1) or plus cisplatin in dose of 0.1 and 1.0 microg x mL(-1). At the same time, SLAP (80 - 320 microg x mL(-1)) also could inhibite the expression of c-myc gene of SGC -7901 cells.

CONCLUSIONSingle SLAP had inhibiting effect on human gastric carcinoma cells proliferation with a dose-effect relationship and synergic effect while combined with adriamycin or cisplatin. To inhibit the expression of c-myc gene of human gastric carcinoma cells might be one of action mechanisms of SLAP, which inhibited tumor cells proliferation.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Dose-Response Relationship, Drug ; Doxorubicin ; pharmacology ; Drug Combinations ; Drug Synergism ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Genes, myc ; Humans ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-myc ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo determine the molecular weight and content of Ginkgo biloba exocarp polysaccharides.

METHODThe analysis was carried on a PL aquagel-OH MIXED (7.5 mm x 300 mm, 8 microm) chromatography column eluted with water as mobile phase at 1.0 mL x min(-1) of flow rate, the column temperature was 25 degrees C and the eluate was detected by RID.

RESULTThe average molecular weight of Ginkgo bilobaexocarp polysaccharides was 11 062.5 with RSD = 0.78% (n = 6); the content was 81.9% with RSD = 2.5% (n = 6), the standard curves of dextran (MW 12 000) were linear in the range of 1-20 microg, r = 0.999 9. The average recovery is 97.9%, RSD was 2.5%.

CONCLUSIONThis method was found to be sensitive and accurate for the measurement of Ginkgo biloba exocarp polysaccharides.

Chromatography, High Pressure Liquid ; methods ; Fruit ; chemistry ; Ginkgo biloba ; chemistry ; Molecular Weight ; Plants, Medicinal ; chemistry ; Polysaccharides ; analysis ; chemistry ; Reproducibility of Results

Objective To validate the performance of droplet digital polymerase chain reaction (ddPCR) reagents using clinical and standard strains, and to evaluate the effectiveness and practicability of ddPCR technology in clinical applications. Methods The concordance rate, specificity, precision, and lower limit of detection of the ddPCR kit were validated using clinical and standard strains. Blood samples from 74 patients with suspected bloodstream infections were collected, and both ddPCR and blood culture methods were used to determine the pathogens in the patient's blood samples. Results The average detection time of ddPCR for pathogens of bloodstream infection was 3.5 hours, which was able to complete the detection of over a dozen common pathogens simultaneously. The concordance rate, specificity, precision, and lower limit of detection of the ddPCR kit for bloodstream infection pathogens all met clinical requirements. Among the 74 patients with suspected bloodstream infections, the positive detection rate using the ddPCR method was 64.86%, while was 40.54% using blood culture, with a statistically significant difference (P < 0.05). Conclusion The method of ddPCR can detect common pathogens of bloodstream infections rapidly, efficiently, and in large quantities, and its main detection performance can meet clinical needs. The combination of blood culture and ddPCR technology is conductive to early and rapid diagnosis of clinical bloodstream infection patients.

Objective To discuss the reason of skin flap necrosis caused by vascular crisis of reverse island flap of forearm posterior interosseous artery.Methods Eight-six patients who were underwent reverse island flap of forearm interosseous posterior artery for deep tissues and skin defect on the back of hand between March,2002 and April,2014 were analyzed in this study.Eleven patients had occurred skin flap necrosis,include 5 cases had completely flap necrosis caused by circulation crisis,and 6 cases had partial necrosis at the distal of the flap.Among the necrosis cases,5 cases were injured by the machine injury,4 cases by the heavy crush and 2 cases by the traffic accident.The cause of circulation crisis was analyzed.Results In the series,75 skin flaps survived completely and 11 cases had occurred necrosis,included completely necrosis with 5 cases.The reasons of flap crisis were as follows:for the completely necrosis,2 cases with variation of perforating branch of posterior interosseous artery,1 case with absence of posterior interosseous artery,1 case with vessel pedicel entrapment in subcutaneous tunnel,and 1 case with misconduct venous congestion caused by the reverse perfusion of superficial vein.The reason of circulation crisis of completely necrosis were as follows:2 cases with artery crisis and 3 of them with distortion of entrapment at pedicel and vein crisis.One case was cured through debridement,change of medical prescription and skin grafting;and 4 cases were cured with other flap repair technique.For the partial necrosis,2 cases with variation of perforating branch of posterior interosseous artery,1 case with excessively narrow entrapment at pedicel in subcutaneous tunnel,1 case with folding vessel pedicel entrapment of skin at the back of wrist,1 case with misconduct of superficial vein trunk and 1 case with intraoperative side-injury.The symptoms of circulation crisis of completely necrosis were as follows:2 cases with artery crisis and 4 of them with distortion of entrapment at pedicel and vein crisis.Four cases were cured through debridement and skin grafting,1 case was cured by the vacuum-sealing drainage (VSD) and 1 case with skin flap repair at pedicle of abdomen.Conclusion The anatomic variation of perforator vessel of reverse island flap of forearm posterior interosseous artery;narrow entrapment at pedicel in subcutaneous tunnel and distortion of entrapment at pedicel;venous congestion caused by the reverse perfusion of superficial vein;intraoperative side-injury of the pedicel of the flap;excessively folding vessel pedicel entrapment of skin at the back of wrist after surgery will cause the circulation crisis of reverse island flap of forearm posterior interosseous artery and induce the necrosis of the skin flap.

This study was purposed to observe the influence of umbilical cord mesenchymal stem cells (UC-MSC) on the peripheral blood CD4(+)CD25(+)regulatory T cells (Treg), Th17 cells and neutrophils in rats with collagen type II-induced arthritis(CIA), and to explore the regulating effect of UC-MSC transplantation on immunocyte subgroup. The rats wee divided into 3 groups: CIA group (model group), UC-MSC treated group and blank control group. The CIA rats were injected with UC-MSC via tail vein. The percentage of CD4(+)CD25(+) cells in peripheral blood and the expression of NCD11b on neutrophil surface in CIA rates was detected by flow cytometry (FCM), and the serum interleukin-17 (IL-17) was observed by enzyme-linked immunosorbent assay (ELISA). The results showed that the mean fluorescence intensity(MFI) of NCD11b and the level of IL-17 in the model group were significantly higher than those in the blank control group, and the ratio of CD4(+)CD25(+) cells were significantly lower (P < 0.05). The MIF of NCD11b and the level of IL-17 in the UC-MSC treated group were significantly lower than that in the model group (P < 0.05), while the proportion of CD4(+)CD25(+) Treg increased (P < 0.05). Since the fifth week, the above indicators in the UC-MSC group have almostly approached the control group. It is concluded that the UC-MSC can increase peripheral blood Treg proportion in CIA rat, inhibit the secretion of Th17 and the activity of neutrophils, reduce the immune inflammation reaction, decrease the release of proinflammatory factor, and induce immune reconstruction.
Animals ; Arthritis, Experimental ; immunology ; therapy ; Female ; Interleukin-17 ; metabolism ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Neutrophils ; immunology ; Rats ; Rats, Sprague-Dawley ; Th17 Cells ; immunology ; Umbilical Cord ; cytology

AIM: To study the chemical constituents of the solid culture of the endophyte Phomopsis sp. IFB-E060 in Vatica mangachapoi. METHOD: Isolation and purification were performed through silica gel column chromatography, gel filtration over Sephadex LH-20, ODS column chromatography, and HPLC. Structures of the isolated compounds were elucidated by a combination of spectroscopic analyses (UV, CD, IR, MS, 1D, and 2D NMR). The cytotoxicity of the isolates was evaluated in vitro by the MTT method against the human hepatocarcinoma cell line SMMC-7721. RESULTS: Five compounds were isolated from the solid culture of the endophyte Phomopsis sp. IFB-E060 and their structures were identified as 18-methoxy cytochalasin J (1), cytochalasin H (2), (22E, 24S)-cerevisterol (3), ergosterol (4), and nicotinic acid (5). Compound 1 had an inhibition rate of 24.4% at 10 μg·mL(-1) and 2 had an IC50 value of 15.0 μg·mL(-1), while a positive control 5-fluorouracil had an inhibition rate of 28.7% at 10 μg·mL(-1). CONCLUSION 18-Methoxy cytochalasin J (1), produced by endophytic Phomopsis sp. IFB-E060, is a new cytochalasin with weak cytotoxicity to the human hepatocarcinoma cell line SMMC-7721.
Ascomycota ; chemistry ; isolation & purification ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cytochalasins ; chemistry ; isolation & purification ; toxicity ; Endophytes ; chemistry ; isolation & purification ; Humans ; Magnoliopsida ; microbiology ; Molecular Structure ; Plant Bark ; microbiology

Objective To observe the effect of Kinesio taping combining with sling exercise therapy (SET) on the motor function for children with spastic hemiplegia. Methods From October, 2014 to April, 2016, 60 children with spastic hemiplegia of cerebral palsy were randomly divided into control group (n=20), SET group (n=20) and observation group (n=20), who accepted routine rehabilitation, routine rehabilitation+SET and routine rehabilitation+SET+Kinesio taping, respectively. They were assessed with Mallet Score, modified Ashworth Scale (MAS), Peabody Developmental Motor Scale-fine motor (PDMS-2FM) and Gross Motor Function Measure (GMFM)-C zone before and three months after treatment. Results The Mallet Score and the scores of PDMS-2FM and GMFM-C improved in all the groups after treatment (t>2.074, P<0.05), and improved the most in the observation group (F>10.032, P<0.001). The score of MAS decreased in the observation group after treatment (t=4.767, P<0.001), and was the least among these groups (F=4.262, P<0.05). Conclusion Kinesio taping combining with SET can improve the motion in the children with spastic hemiplegia, and promote the development of both fine and gross motor function.

Objective To develop a radiomics nomogram based on contrast-enhanced CT (CECT) for preoperative prediction of high-risk and low-risk thymomas. Methods Clinical data of patients with thymoma who underwent surgical resection and pathological confirmation at Northern Jiangsu People's Hospital from January 2018 to February 2023 were retrospectively analyzed. Feature selection was performed using the Pearson correlation coefficient and least absolute shrinkage and selection operator (LASSO) method. An ExtraTrees classifier was used to construct the radiomics signature model and the radiomics signature. Univariate and multivariable logistic regression was applied to analyze clinical-radiological characteristics and identify variables for developing a clinical model. The radiomics nomogram model was developed by combining the radiomics signature and clinical features. Model performance was evaluated using area under the curve (AUC), sensitivity, specificity, accuracy, negative predictive value, and positive predictive value. Calibration curves and decision curves were plotted to assess model accuracy and clinical values. Results A total of 120 patients including 59 females and 61 males with an average age of 56.30±12.10 years. There were 84 patients in the training group and 36 in the validation group, 62 in the low-risk thymoma group and 58 in the high-risk thymoma group. Radiomics features (1 038 in total) were extracted from the arterial phase of CECT scans, among which 6 radiomics features were used to construct the radiomics signature. The radiomics nomogram model, combining clinical-radiological characteristics and the radiomics signature, achieved an AUC of 0.872 in the training group and 0.833 in the validation group. Decision curve analysis demonstrated better clinical efficacy of the radiomics nomogram than the radiomics signature and clinical model. Conclusion The radiomics nomogram based on CECT showed good diagnostic value in distinguishing high-risk and low-risk thymoma, which may provide a noninvasive and efficient method for clinical decision-making.

BACKGROUNDWhite matter lesions (WMLs) are common findings in brain magnetic resonance imaging (MRI) and are strongly associated with stroke incidence, recurrence, and prognosis. However, the relationship between WMLs and transient ischemic attacks (TIAs) is not well established. This study aimed to determine the clinical significance of WMLs in patients with TIA.

METHODSA total of 181 consecutive inpatients with first-ever TIA were enrolled. Brain MRIs within 2 days of symptom onset were used to measure WML volumes. Recurrent vascular events within 1 year of TIA onset were assessed. The relationship between WMLs and recurrent risk of vascular events was determined by a multivariate logistic regression.

RESULTSWMLs were identified in 104 patients (57.5%). Age and ratio of hypertension were significantly different between patients with and without WMLs. The incidence of vascular events in patients with WMLs significantly increased in comparison to those without WMLs (21.15% vs. 5.19%, 95% confidence interval [CI]: 1.18-15.20, P = 0.027) after controlling for confounders. Furthermore, distributions of WML loads were found to be different between patients who developed vascular events and those who did not. WML volumes were demonstrated to be correlated with recurrent risks, and the fourth quartile of WML volumes led to an 8.5-fold elevation of recurrent risk of vascular events compared with the first quartile (95% CI: 1.52-47.65, P = 0.015) after adjusting for hyperlipidemia.

CONCLUSIONWMLs occur frequently in patients with TIA and are associated with the high risk of recurrent vascular events, suggesting a predictive neuroimaging marker for TIA outcomes.