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 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

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

Objective To investigate the expression and clinical significance of microRNA-148a-3p (miR-148a-3p) in lung adenocarcinoma and analyze the effect and mechanism of miR-148a-3p on radiotherapy sensitivity of lung adenocarcinoma cells by targeting the protein core 1β13-galactosyltransferase 1(C1GALT1). Methods Seventy-six patients' tumor tissues from lung adenocarcinoma tissue microarrays and lung adenocarcinoma A549 cell line were selected for the study. The miR-148a-3p in situ hybridizations (ISH) and C1GALT1 immunohistochemical staining were performed on the tissue microarrays to analyze the correlations of miR-148a-3p expression with clinical pathology, prognosis and C1GALT1 expression in the tumor tissues of the 76 patients with lung adenocarcinoma. A549 cells were transfected with miR-148a-3p overexpression plasmid by using cell transfection technique; the clone formation assay was used to detect the sensitivity of the transfected cells for radiotherapy after receiving 2 Gy radiotherapy; the protein expression level of cellular C1GALT1 was detected by western blot; the targeted regulatory relationship between miR-148a-3p and C1GALT1 was verified by dual-luciferase reporter gene experiment; the mechanism of miR-148a-3p regulating the sensitivity of A549 cells to radiotherapy was analyzed by co-transfection technique. Results Low expression of miR-148a-3p in 76 cases of lung adenocarcinoma tissues was significantly associated with lymph node metastasis (P=0.012); the prognosis of patients with high expression of miR-148a-3p was significantly better than that of patients with low expression of miR-148a-3p (P=0.005); there was a significant negative correlation between the expression of miR-148a-3p and C1GALT1 in lung adenocarcinoma tissues (P=0.023). Multivariate analysis showed that low expression of miR-148a-3p, T staging and lymph node metastasis were the independent risk factors for prognosis. Clone formation experiment showed that the number of clone formation in the miR-148a-3p overexpression group was significantly lower than that in the control group (P < 0.001); western blot result showed that overexpression of miR-148a-3p was able to significantly down-regulate the level of C1GALT1 protein in cells (P < 0.001). Dual luciferase reporter gene experiment showed that miR-148a-3p was able to regulate the expression of C1GALT1 by binding to the 3'UTR of C1GALT1. Functional rescue experiment showed that C1GALT1 could partially offset the radiosensitization effect of miR-148a-3p. Conclusion Low expression of miR-148a-3p in lung adenocarcinoma is significantly associated with lymph node metastasis, high expression of C1GALT1 and poor prognosis, and miR-148a-3p can enhance the radiosensitivity of lung adenocarcinoma cells by negatively regulating the expression of C1GALT1.

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

Pan-immune inflammation value (PIV) is a comprehensive immune inflammatory biomarker based on complete blood cell counts, which has been proven to predict treatment response and survival outcomes for different types of tumors. However, the predictive value of the PIV varies in different strategies for tumor treatment. This paper aims to systematically review the latest progress of PIV in predicting survival outcomes and tumor prognosis for immunotherapy, radiotherapy, targeted therapy, endocrine therapy, surgical treatment and neoadjuvant therapy, and analyze its existing challenges and issues, as well as look forward to its future development direction and application prospects.

Objective To investigate the correlation between pre-treatment pan-immune inflammation value (PIV) and clinicopathological features in esophageal squamous cell carcinoma (ESCC) patients with postoperative adjuvant radiotherapy and evaluate its value in prognosis assessment combined with T stage. Methods A retrospective analysis was conducted on data of 85 ESCC patients with postoperative adjuvant radiotherapy in the Department of Radiation Oncology of the Affiliated Hospital of Yangzhou University from January 2019 to January 2023. The receiver operating characteristic (ROC) curve was drew to obtain the optimal cut-off value of PIV and other immune-inflammatory biomarkers. The area under the curve (AUC) and clinical applicability of PIV and other immune-inflammatory biomarkers were compared based on the ROC curve and decision curve analysis (DCA). According to the optimal cut-off value, patients were divided into high PIV group and low PIV group, and the correlation between PIV level and clinicopathological features of ESCC was evaluated. Kaplan-Meier method was used for survival analysis, the Cox proportional hazards model was used for multivariate analysis, and a risk stratification model combining PIV and T stage was established by recursive partitioning analysis (RPA). Results The optimal cut-off value of pre-treatment PIV was determined as 187.22 based on the ROC curve. The AUC of PIV was 0.679, which was greater than 0.640, 0.583, 0.656 and 0.644 of the other four immune-inflammatory biomarkers such as the systemic immune-inflammation index (SII), platelet-to-lymphocyte ratio (PLR), monocyte-to-lymphocyte ratio (MLR), and neutrophil-to-lymphocyte ratio (NLR). The 85 patients were divided into low PIV group (< 187.22, n=48) and high PIV group (≥187.22, n=37). The level of PIV was significantly correlated with tumor diameter (P < 0.05). The 3-year overall survival (OS) (75.6% versus 30.6%, P < 0.001) and 3-year disease-free survival (DFS) (56.1% versus 21.0%, P < 0.001) were significantly higher in the low PIV group than the high PIV group. Tumor diameter, T stage and PIV were independent factors affecting OS in ESCC patients (P < 0.05), and T stage and PIV were independent factors affecting DFS in ESCC patients (P < 0.05). A new staging system with three risk groups was established by the RPA stratification model based on T stage and PIV, which further improved the predictive value of prognosis compared with T stage or PIV alone. Conclusion Pre-treatment PIV is helpful in predicting the prognosis of ESCC patients with postoperative adjuvant radiotherapy, and the combination of PIV and T stage can improve the predictive value.

Early postoperative inflammatory small bowel obstruction is one of the complications after abdominal surgery. Most obstructions will resolve on their own after nutritional support. However, the recovery of intestinal function takes a long time, which leads to high costs and is prone to occur related complications. Early postoperative inflammatory small bowel obstruction is also one of the risk factors for secondary adhesive intestinal obstruction. A few patients with ineffective conservative treatment have to undergo high-risk surgical treatment. Cases of early postoperative inflammatory small bowel obstruction complicated by superior mesenteric artery syndrome are relatively rare. Comprehensive treatment including acupuncture, hormones, and water-soluble contrast agents as well as early placement of enteral nutrition tubes is the key to the treatment of such patients.