Biomedical Research ; Humans ; Mouth ; radiation effects ; Radiation Injuries ; therapy

OBJETCTIVE:To establish a method for the content determination of ethacridine lactate in Compound ethacridine ointment. METHODS:HPLC was performed on the column of Agilent ZORBAX SB-C18 with mobile phase of 0.1% Octanesulfon-ic acid sodium solution-acetonitrile(70∶30,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 270 nm,the column temperature was 30℃,and the injection volume was 10 μl. RESULTS:The linear range of ethacridine lactate was 10.002-50.010μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were less than 1%;recovery was 98.96%-100.36%(RSD=0.49%,n=9). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for content determina-tion of ethacridine lactate in Compound ethacridine ointment.

Objective:To summarize the experience of Bacillus Calmette-Guerin(BCG) in the treatment of bladder cancer secondary to renal transplantation.Methods:The clinical data of 5 patients who underwent BCG bladder irrigation after secondary bladder cancer after kidney transplantation in Tianjin First Central Hospital from January 2015 to December 2019 were analyzed. There were 1 male and 4 female cases. During the period of immunosuppression after transplantation, 1 case developed secondary high-level non-muscular invasive bladder cancer (NMIBC), 3 cases developed secondary low-grade NMIBC, and 1 case developed secondary glandular cystitis (4 cases). The mean age of the 5 patients with secondary bladder cancer was 59.7±4.0 years. Case one with high level NMIBC was treated with transurethral resection of bladder tumor (TURBT) and postoperative irrigation of epirubicin. Case 3 and 5 with low-level NMIBC accepted regular postoperative irrigation of gemcitabine. No irrigative therapy was performed in case 2. Bladder cancer recurred in case 1, 2, 3 and 5 after 20.1±9.7 months. TURBT was observed in all the 4 patients, among which 3 were of high grade NMIBC and 1 was of low grade NMIBC. Four patients were irrigated with BCG 2 weeks after operation. Postoperative pathology indicated low-level NMIBC in case 4, and BCG was irrigated 2 weeks after the operation. During perfusion therapy, immunosuppressive agents were continued.Results:During BCG perfusion, 4 of the 5 cases showed BCG related local inflammation, among which 2 cases presented symptoms of bladder irritation, 1 case presented hematuria, and 1 case presented hematuria with low fever. Patients with frequent urination, pain in urine, hematuria and other symptoms improved after drinking plenty of water, taking bed rest and taking levofloxacin (0.5g/ day ×7 days). Patients with low fever were treated with antipyretic treatment. No antituberculous agents were used prophylactically during BCG perfusion. There were no symptoms of tuberculosis infection or sepsis. The function of transplantated kidney was normal and no tendency of rejection. The 5 patients were followed up for 7-24 months, 1 patient was lost to follow-up after 7 months of BCG bladder perfusion, and no tumor recurrence or metastasis was found in 5 patients during the follow-up.Conclusions:The use of immunosuppressive agents does not reduce the biological activity of BCG, and BCG does not increase the risk of systemic toxicity or affect the function of transplanted kidneys in immunocompromised patients. BCG is a treatment option for bladder cancer secondary to renal transplantation.

Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.

This study was aimed to evaluate the effect of region of interest (ROI) on measurement of apparent diffusion coefficient (ADC) obtained in diffusion-weighted (DW) MR sequences between spinal tuberculosis and malignancy, selecting the more reasonable and effective measurement of ADC value. We collected the conventional MR sequences and diffusion-weighted imaging data of thirty-two patients with spinal tuberculosis or malignancy confirmed clinically and pathologically from August 2009 to June 2010. Using the conventional MRI as a guide, the signal intensity (SI) and ADC value were calculated by selecting single ROI and the same area multi-ROI within whole lesion at the largest slice of lesion respectively. Using ADC value defined by receiver operating characteristic (ROC) curve analysis as threshold, the sensitivity, specificity, and accuracy of ADC value for diagnosing spinal tuberculosis were compared. The results showed that the mean of ADC value at signal ROI was lower than that at the multi-ROI. There was no significant difference between the ADC value of single ROI and multi-ROI. The ADC values from spinal tuberculosis and spinal malignancy obtained in single ROI or multi-ROI showed significant differences. The sensitivity, specificity, and accuracy of ADC value for differentiating tuberculosis from neoplastic infiltration at signal ROI is lower than that at the multi-ROI. In conclusion, the ADC acquired multi-ROI can describe preferablywater molecular diffusion and is more useful in differentiating spinal tuberculosis from malignant bone marrow lesions.
Adult ; Algorithms ; Diagnosis, Differential ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Image Interpretation, Computer-Assisted ; Male ; Middle Aged ; Spinal Neoplasms ; diagnosis ; Tuberculosis, Spinal ; diagnosis

Objective:To evaluate the role of TANK-binding kinase-1 (TBK1) in renal fibrosis in mice with acute kidney injury (AKI) and relationship with endoplasmic reticulum stress.Methods:Twenty-four male wild-type C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group CON), group AKI, control plus TBK1 inhibitor group (group CON-GSK) and AKI plus TBK1 inhibitor group (group AKI-GSK). In group AKI and group AKI-GSK, folic acid 250 mg/kg was intraperitoneally injected to prepare AKI model.From the first day after folic acid injection, 1% dimethyl sulfoxide 20 ml/kg was intraperitoneally injected every other day in group AKI, and GSK8612 1.5 mg/kg was intraperitoneally injected every other day in group AKI-GSK, 7 times in total.In group CON and group CON-GSK, 1% dimethyl sulfoxide 20 ml/kg and GSK8612 1.5 mg/kg were intraperitoneally injected, respectively, every other day for 7 times in total.On the 14th day after injection of folic acid, the eyeball blood samples were taken to determine the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr), and the kidney tissues were also extracted, and the pathological results of renal tissue were observed by Sirius red staining, Masson staining and HE staining.The area of renal fibrosis was measured and the tubulointerstitial injury score was calculated.The expression of fibronectin, type I collagen and α-smooth muscle actin was detected by immunofluorescence.The expression of phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), activated transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), phosphorylated inositol-requiring kinase 1α (p-IRE1α), tumor necrosis factor receptor-associated factor 2 (TRAF2), apoptosis-regulating signal kinase 1 (ASK1), caspase-12 and phosphorylated c-Jun N-terminal protein kinase (p-JNK) was detected by Western blot. Results:Compared with group CON, the serum BUN and Cr concentrations, area of renal fibrosis and renal tubulointerstitial injury score were significantly increased, and the expression of fibronectin, type I collagen, α-smooth muscle actin, p-PERK, p-eIF2α, ATF4, CHOP, p-IRE1α, TRAF2, ASK1, caspase-12, and p-JNK was up-regulated in group AKI and group AKI-GSK ( P<0.05), and no significant change was found in the indexes mentioned above in group CON-GSK ( P>0.05). Compared with group AKI, the serum BUN and Cr concentrations, area of renal fibrosis, and tubulointerstitial injury score were significantly decreased, and the expression of fibronectin, type I collagen, α-smooth muscle actin, p-PERK, p-eIF2α, ATF4, CHOP, p-IRE1α, TRAF2, ASK1, caspase-12, and p-JNK was down-regulated in group AKI-GSK ( P<0.05). Conclusions:TBK1 is involved in the process of renal fibrosis in mice with AKI, and the mechanism may be related to the promotion of endoplasmic reticulum stress.

Objective:To explore the factors influencing prognosis of patients with in-hospital cardiac arrest (IHCA).Methods:A retrospective observational study was conducted. The clinical data of patients who developed IHCA and underwent cardiopulmonary resuscitation (CPR) at the Second Xiangya Hospital of Central South University from January 1, 2016, to December 31, 2022 were analyzed. The patients' information, including gender, age, medical history, pre-cardiac arrest related parameters [1-hour pre-cardiac arrest neurological function, 24-hour pre-cardiac arrest hemoglobin (Hb) levels, 1-hour pre-cardiac arrest vital signs], initial CPR-related factors (implementation time and location, initial rhythm, ventilation method, defibrillation and resuscitation drugs) as well as restoration of spontaneous circulation (ROSC) related parameters (vital signs at ROSC and 1 hour after ROSC, 24-hour post-cardiac arrest Hb, and IHCA events), were collected through the hospital's electronic medical record system. The clinical data were compared between ROSC and non-ROSC patients as well as between patients with favorable neurological function [cerebral performance category (CPC) grades 1-2] and unfavorable neurological function (CPC grades 3-5) at 28 days. The factors with statistical significance in univariate analysis and clinical significance were enrolled in a binary multivariate Logistic regression model to analyze the influencing factors of ROSC and neurological function at 28 days after ROSC. The predictive value of factors influencing neurological function at 28 days was assessed using receiver operator characteristic curve (ROC curve).Results:A total of 277 IHCA-CPR patients were enrolled, of which 230 achieved ROSC (83.0%) and 47 were not achieved (17.0%). Compared with non-ROSC patients, ROSC patients had lower prevalence of cerebrovascular disease history and proportion of adrenaline usage, but a higher proportion of initial shockable rhythms. In the multivariate Logistic regression analysis, it was found that using a bag-mask ventilation+endotracheal intubation (compared with a bag-mask ventilation alone) was beneficial for achieving ROSC in IHCA-CPR patients [odds ratio ( OR) = 2.895, 95% confidence interval (95% CI) was 1.204-6.962, P = 0.018], while a initial non-shockable rhythm was not conducive to achieving ROSC in IHCA-CPR patients ( OR = 0.349, 95% CI was 0.147-0.831, P = 0.017). Among the 230 ROSC patients, 42 had good neurological function at 28 days (18.3%), and 188 had poor neurological function (81.7%). Compared with the patients with good neurological function, the patients with the poor neurological function were older and had a higher prevalence of 1-hour pre-cardiac arrest neurological dysfunction and low perfusion, initial non-shockable rhythms, endotracheal intubation, and usage of adrenaline, vasopressors and sodium bicarbonate, a lower proportion of defibrillation and antiarrhythmic medication usage as well as lower 24-hour post-cardiac arrest Hb levels. The multivariate Logistic regression analysis revealed that female ( OR = 6.449, 95% CI was 1.837-22.642, P = 0.004), older age ( OR = 1.054, 95% CI was 1.017-1.093, P = 0.004), 1-hour pre-cardiac arrest neurological dysfunction ( OR = 25.044, 95% CI was 2.737-229.169, P = 0.004), 1-hour pre-cardiac arrest low perfusion ( OR = 3.880, 95% CI was 1.306-11.524, P = 0.028), endotracheal intubation (compared with a bag-mask ventilation; OR = 8.712, 95% CI was 1.402-54.141, P = 0.020) and face mask+endotracheal intubation during CPR (compared with a bag-mask ventilation; OR = 11.089, 95% CI was 3.482-35.320, P = 0.000), IHCA events > 1 time ( OR = 4.221, 95% CI was 1.249-14.226, P = 0.020) were positively associated with poor neurological function at 28 days, which were independent risk factors those were not conducive to 28-day neurological function recovery after ROSC in IHCA-CPR patients. In contrast, usage of antiarrhythmic medication ( OR = 0.345, 95% CI was 0.134-0.890, P = 0.028) and 24-hour post-cardiac arrest Hb ( OR = 0.983, 95% CI was 0.966-0.999, P = 0.043) were negatively associated with poor neurological function at 28 days, which were protective factors those were beneficial for the recovery of neurological function. ROC curve analysis showed that the area under the ROC curve (AUC) of 24-hour post-cardiac arrest Hb for predicting poor neurological function at 28 days after ROSC in IHCA-CPR patients was 0.659 (95% CI was 0.577-0.742), with a cut-off value of 99.5 g/L (sensitivity was 76.2%, specificity was 57.8%). Conclusions:Defibrillation and tracheal intubation during CPR are crucial for IHCA patients. It was also observed that patients with low Hb (< 99.5 g/L should be of high concern), older age, 1-hour pre-cardiac arrest neurological function and hypoperfusion, and IHCA events > 1 time were significantly related to unfavorable neurological outcome in adult resuscitated patients with IHCA.

Objective To investigate the characteristics of TCM constitution distribution in hepatitis B cirrhosis patients with dysplastic nodules(DN),and to provide a basis for the prevention and treatment of precancerous lesions of liver cancer.Methods This study was conducted among 113 hepatitis B cirrhosis patients with DN,105 hepatitis B cirrhosis patients with regenerative nodules(RN),and 70 hepatitis B cirrhosis patients with small hepatocellular carcinoma(sHCC)who were hospitalized in Guangdong Provincial Hospital of Traditional Chinese Medicine from May 2015 to March 2023.Related data were collected,including age,sex,liver function Child-Pugh class,TCM constitution type,and laboratory markers.A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups;the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups;the chi-square test was used for comparison of categorical data between groups,and the Bonferroni correction method was used for further comparison between two groups.Results The main TCM constitution types of hepatitis B cirrhosis patients with DN were Qi-deficiency constitution(27 patients,23.89%),blood-stasis constitution(26 patients,23.01%),and phlegm-dampness constitution(23 patients,20.35%).There were significant differences between the three groups in the proportion of patients with phlegm-dampness constitution or damp-heat constitution(χ2=6.822 and 6.383,both P<0.05);the hepatitis B cirrhosis patients with RN had the highest proportion of patients with phlegm-dampness constitution(30.48%),followed by those with DN(20.35%)and those with sHCC(14.29%),while the hepatitis B cirrhosis patients with sHCC had the highest proportion of patients with damp-heat constitution(27.14%),followed by those with DN(16.81%)and those with RN(12.38%).There were significant differences between the hepatitis B cirrhosis DN patients with different TCM constitution types in sex,age,Child-Pugh class,prealbumin,albumin(Alb),aspartate aminotransferase,total bilirubin(TBil),total bile acid,and alpha-fetoprotein(all P<0.05).Compared with the male hepatitis B cirrhosis DN patients,female patients showed a significantly higher proportion of patients with Qi-deficiency constitution(χ2=4.895,P=0.027).Among the patients with Qi-deficiency constitution,the patients with Child-Pugh class A liver function accounted for a significantly lower proportion than those with Child-Pugh class B liver function(χ2=6.380,P=0.012),while among the patients with phlegm-dampness constitution,the patients with Child-Pugh class A liver function accounted for a significantly higher proportion than those with Child-Pugh class B liver function(χ2=8.515,P=0.004).The patients with phlegm-dampness constitution had significantly higher levels of prealbumin and Alb than those with the other four constitutions(all P<0.05),as well as significantly lower levels of TBil and total bile acid than those with damp-heat constitution(P<0.05);the patients with Yin-deficiency constitution had a significantly lower level of Alb than those with qi-deficiency constitution,blood-stasis constitution,or phlegm-dampness constitution(all P<0.05);the patients with Yin-deficiency constitution had a significantly lower proportion of patients with abnormal alpha-fetoprotein than those with non-Yin-deficiency constitutions(χ2=4.448,P=0.035).Conclusion Hepatitis B cirrhosis patients with DN mainly have the TCM constitution types of Qi deficiency,blood stasis,and phlegm dampness.The patients with phlegm-dampness constitution seem to have a low probability of carcinogenesis,while those with damp-heat constitution and Yin-deficiency constitution have a relatively high risk of carcinogenesis.

Objective To investigate the optimal digestion method for the preparation of single-cell suspensions from mouse small intestinal lamina propria.Methods Ten mouse small intestines of uniform length were collected and randomly divided into two groups.Each group was used to prepare lamina propria single-cell suspensions by enzymatic digestion with collagenase A or collagenase Ⅷ.We compared the effects of these two enzymatic digestion method on the cell yield,cell activity,and cell surface antigens of the single-cell suspensions.Single-cell suspensions prepared by the superior enzymatic digestion method were then subjected to flow cytometry assay.Results Compared to collagenase-Ⅷ-based enzymatic digestion,collagenase-A-based digestion result ed in a higher cell yield(9.48±1.10)× 109vs(4.18±1.02)×109and higher proportions of live cells(86.36±3.32)％vs(61.62±10.93)％,active CD45+cells(57.19±5.11)％vs(26.01±11.44)％,active CD3+cells(8.73±2.89)％vs(4.52±2.49)％,active CD4+cells(6.19±2.09)％vs(3.22±1.91)％,and active B220+cells(15.06±4.27)％vs(5.07±2.20)％,providing high-quality cells for subsequent flow assays.Conclusions The collagenase A-based enzymatic digestion method is more suitable for the preparation of ingle-cell suspensions from the lamina propria of ouse small intestines.

Purpose: The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT. Materials and Methods: This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles. Results: During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively. Conclusion Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.