Objective:To optimize the extraction technology of Fufang Jinsha Lidan granule. Methods:According to the properties of traditional Chinese medicines in the formula, orthogonal experiments were used to optimize the extraction process of Fufang Jinsha Li-dan granule. The volume of water, boiling time and boiling frequency were used as the three influencing factors with three different lev-els in the orthogonal experiments. Moreover, the content of paeoniflorin and the yield of extract were chosen as the evaluation indices. The orthogonal experiments were carried out according to the L9 (34 ) orthogonal table. Results:The optimal extraction process of Fu-fang Jinsha Lidan granule was as follows:boiled twice, and one hour per time with 12-fold amount of water ( soaked for 30 min with 14-fold amount of water for the first extraction process) . Conclusion:The extraction process has such properties as high extraction rate, stability, simple operation, high yield of extract and controllable quality, which is worthy of wide application.

Objective To evaluate the safety of morphine hydrochloride injection. Methods Ear verin injection was used to evaluate the vascular irritation using the comparison of left side with right side in rabbits. Quadriceps femoris injection was used to evaluate the muscle irritation using the comparison of left side with right side in rabbits. Guinea pigs were intravenously injected with morphine hydrochloride injection at a dose of 2.8 mg·kg-1 once daily 3 times, stimulation was performed on 14 d after the last sensitization and the booster dose was 2 times the sensitization dose. The allergic reactions were observed. The different concentrations of morphine hydrochloride injection were placed in 2％ rabbit erythrocyte suspension, and then the hemolyzation and agglutination were observed. Results There were no significant vascular or muscular irritation and injury effects of morphine hydrochloride injection in rabbits. There were no evidenceof hemolyzation and agglutination in rabbit erythrocytes in vitro. No allergic reactions on guinea pigs in vivo were observed. Conclusion After treatment of morphine hydrochloride injection, neither obvious vascular /muscle stimulation or sensitization, nor hemolyzation or agglutination appeared in rabbits. The research results provide basic reference for the clinical rational and safe application of morphine hydrochloride injection.

Objective To explore the diagnostic value of dynamic contrast-enhanced MRI (DCE-MRI) in evaluation of hemodynamic parameters of lesions and background parenchyma in benign and malignant breast lesions.Methods Breast DCE-MRI data were collected from 41 patients,including 21 benign lesions (benign group) and 24 malignant lesions (malignant group).Volume transfer constant (K),rate constant (Kep) and plasma fraction (Vp) of lesions and background parenchyma were measured,and the parameters of both groups were statistically analyzed.Taking pathological diagnosis as the gold standards,the efficacy of parameters in diagnosis of malignant breast lesions was evaluated.Results Ktrans and Kep of lesions and background parenchyma in the malignant group were higher than those in benign group (all P＜0.05).There was no significant difference between Vp of lesions and background parenchyma in both groups (both P＞0.05).In the diagnosis of malignant lesions,AUC of Ktrans of background parenchyma was the best,the sensitivity and specificity was 87.50％ and 76.19％,respectively.All parameters combined Logistic regression model had the highest AUC value (0.86,P＜0.001).Conclusion Among DCE-MRI related parameters,K of background parenchyma has the highest diagnostic value.Quantitative analysis of DCE-MRI data can help to identify benign and malignant breast lesions,and the hemodynamic characteristics of the background parenchyma are expected to be new methods for non-invasive diagnosis of breast lesions.

OBJECTIVEThrough analysis of variation and function of 5 main endogenous hormones in the formation of microtuber of Dioscorea opposite in vitro to explore the physiological and biochemical mechanism of microtuber development.

METHODWhen microtubers were induced on MS + 6-BA 1.5 mg x L(-1) + NAA 1.5 mg x L(-1) + sucrose 5% medium, the endogenous hormones were isolated during different formation stages of microtubers, then purified and detected with enzyme-linked immunosorbent assays (ELISA).

RESULTThe results showed that GA3 slightly decreased in initial period, rose suddenly 20 days later, and than decreased. IAA showed a dropping tendency in the total course, ABA and ZR increased in a long period, dropped at last. JA continuously rose and never dropped, GA3 and ABA and the ratio of GA3 and JA varied obviously.

CONCLUSIONIAA, ABA, JA , ZR and GA3 play an important role in controlling formation of microtubers in D. opposite in vitro.

Abscisic Acid ; metabolism ; Cyclopentanes ; metabolism ; Dioscorea ; growth & development ; metabolism ; Gibberellins ; metabolism ; Indoleacetic Acids ; metabolism ; Isopentenyladenosine ; analogs & derivatives ; metabolism ; Oxylipins ; metabolism ; Plant Growth Regulators ; metabolism ; Plant Tubers ; growth & development ; metabolism

Objective: To investigate the therapeutic effects of compound Kushen Tang and its relevant mechanism in TNBS-in-duced ulcerative colitis ( UC) rats. Methods:UC was induced by TNBS in rats. After compound Kushen Tang was given orally, the levels of MDA, iNOS, and NO and the activity of MPO, SOD, and GSH-Px were measured. The general condition of rats and colon tissue morphology were observed. Results:The levels of MDA (P<0. 05), iNOS (P<0. 01) and NO (P<0. 01) and the activity of MPO (P<0. 01) in tissues of UC rats were significantly higher than the control group. The activity of SOD (P<0. 01) and GSH-Px (P<0. 05) were significantly lower than those in the control group. After the treatment with high doses of compound Kushen Tang, the levels of MPO (P<0. 01), MDA (P<0. 05), iNOS (P<0. 05) and NO (P<0. 01) were significantly decreased, and the activity of SOD (P<0. 01) and GSH-Px (P<0. 05) significantly increased. The therapeutic effect was dose-dependent and the general con-dition of rats and colon tissue morphology were also significantly improved. Conclusion:Compound Kushen Tang is considered as a no-vel therapeutic alternatives for the treatment of UC, which can reduce coloni inflammatory injury and ameliorate the colitis.

Magnetic resonance diffusion tensor imaging has been developed to a kind of new imaging technology in recent years;through the nerve tissue diffusion properties of water molecules,it can quantitatively reflect the integrity of white matter fiber tracts and various kinds of pathological changes,thus achieving accurate evaluation of spinal cord injuries at the micro level.In this paper,the technology of magnetic resonance diffusion tensor imaging in the application of electroacupuncture treatment of spinal cord injuries and research progress are summarized and prospected.

Objective:To investigate the predictive value of diffusion tensor imaging (DTI) parameters in upper extremity motor function recovery after surgery in patients with acute cervical spinal cord injury (CSCI).Methods:Twenty-three patients with acute CSCI who received postoperative systemic rehabilitation therapy in Department of Neurosurgery, 900 th Hospital of Joint Logistics Team of People's Liberation Army from May 2019 to July 2021 were selected as an experimental group, and 22 healthy subjects (healthy control group) matched with age and gender were selected from Physical Examination Center of the same hospital at the same period. Routine MRI sequence and DTI scan of the cervical spinal cord, scale of American Association for Spinal Cord Injury (ASIA) and modified Barthe index (mBI) were performed in patients of the experimental group 1 d and 3 months after surgery. Routine MRI sequence and DTI scan of the cervical spinal cord were performed in healthy subjects after enrollment. The DTI parameters in different regions between the two groups were compared, and the differences in DTI parameters, ASIA scores and mBI in patients of the experimental group before and after surgery were compared. Correlations of preoperative DTI parameters with preoperative upper extremity motor ASIA scores and upper extremity motor recovery rate 3 months after surgery were analyzed by Pearson correlation analysis. Receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of preoperative fractional anisotropy (FA) in upper extremity motor function recovery in CSCI patients 3 months after surgery. Results:As compared with the healthy control group, the experimental group had significantly lower preoperative FA in the injury area and distal injury area, and statistically higher preoperative apparent diffusion coefficient (ADC, P<0.05). In patients of the experimental group, preoperative FA in the injury area was significantly lower and ADC in the injury area was significantly higher as compared with those in the distal injury area ( P<0.05); patients of the experimental group had significantly higher FA in these two regions, upper extremity motor ASIA scores and mBI, and significantly lower ADC 3 months after surgery as compared with those 1 d before surgery ( P<0.05). The preoperative FA in the injury area and distal injury area in CSCI patients were positively correlated with preoperative upper extremity motor ASIA scores and upper extremity motor recovery rate 3 months after surgery ( P<0.05). ROC curve results showed that the area under the curve (AUC) of preoperative FA in injury area in predicting upper extremity motor function recovery 3 months after surgery was 0.912 ( 95%CI: 0.783-1.000, P<0.001); that of preoperative FA in the distal injury area was 0.842 ( 95%CI: 0.682-1.000, P<0.001). Conclusion:DTI parameters FA and ADC are sensitive indicators for detecting CSCI; preoperative FA in the injury area and distal injury area can be used to predict the upper extremity motor function recovery, but the efficacy of the former is superior to that of the later.

Objective To investigate the effect of ERH gene knockdown on the proliferation and apoptosis of human bladder cancer T24 cells. Methods T24 cells infected by lentivirus with interference on ERH gene sequence were cloned to establish stable T24 cells clone in ERH gene suppression. The expression of ERH mRNA gene in bladder cancer was detected by using quantitative real time polymerase chain reaction (qPCR). The effects of ERH knockout on the cell proliferation and apoptosis were examined by using methylthiazolyl tetrazolium (MTT) assay, colony formation assay and flow cytometry. The effect of ERH knockout on the tumorigenic effect of T24 cells in vivo was verified by subcutaneous tumor formation in nude mice. Results After lentiviral transfection, qPCR results showed that the knockdown effect of ERH mRNA in ERH normal group (untreated T24 cells) was better than that in ERH gene knockdown group, and the difference was statistically significant [(1.006±0.126) vs. (0.079±0.007); t=12.72, P=0.0002]. After knocking out ERH gene, MTT assay showed that the proliferation ability of T24 cells in ERH gene knockdown group was weakened compared with ERH normal group, and the difference was statistically significant [A490 value: (0.13±0.00) vs. (0.66±0.01);t=104.61, P<0.0001]. Colony formation assay indicated that the ability of clone in ERH normal group was weakened compared with ERH gene knockdown group [(10.5 ±1.2) vs. (196.4 ±4.0); t= 73.63, P< 0.0001]. Flow cytometry showed that the cell apoptosis rate in ERH gene knockdown group was higher than that in ERH normal group [(11.0 ±0.5) ％ vs. (4.2 ±0.5) ％; t= 16.06, P<0.0001]. Imaging results of subcutaneous tumor formation in nude mice showed that the total fluorescence intensity of the tumor area in ERH gene knockdown group was (4.67 ±0.59) × 1010 μW/cm2, and the corresponding part in ERH normal group was (9.54±4.20) × 1010μW/cm2 (t=3.64, P=0.0051);tumor weight in ERH gene knockdown group was (0.80±0.62) g, and in ERH normal group was (1.79±0.71) g (t=3.33, P=0.0037). Conclusion ERH gene knockout can inhibit the proliferation of human bladder cancer T24 cells, and promote the cell apoptosis.

Objective:To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment. Methods:125I-RSOAds-hTERT/PSA ( 125I-virus complex) oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation. TUNEL staining, flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo, respectively. To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels, interleukin 2 (IL-2), IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the culture supernatant of human prostate cancer cell line PC3, mouse prostate adenocarcinoma cell line RM-1, and mice serum were detected by ELISA. We explored the regulation of 125I-virus complex on the expression of CD24, CD44 and prostate stem cell antigen (PSCA) in prostate tumor tissues and tumor cells through immunohistochemistry. Meanwhile, the expression levels of CD32 and vascular endothelial growth factor (VEGF), as well as CD4+ , CD8+ and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments. Results:125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group. Meanwhile, IL-2 ( t=-183.30, -38.20, P<0.05), IL-10 ( t=113.80, 92.71, P<0.05), TNF-α ( t=-73.20, -73.91, P<0.05), IFN-γ ( t=-65.37, -139.70, P<0.05) increased in vitro and in vivo. 125I-virus complex reduced the expression of CD24, CD44 and PSCA in tumor cells and tumor tissue, reduced the weight of tumor tissue, inhibited angiogenesis of tumor tissue ( t=8.55, P<0.05), and regulated the immune response in tumor tissue. Conclusions:125I-virus complex targeting prostate cancer can significantly kill cancer cells, reduce the weight and angiogenesis of tumor, and improve tumor microenvironment.

Single-cell sequencing is a new technology for high-throughput sequencing analysis of the genome, transcriptome, and epigenome at the single-cell level. Currently, single-cell sequencing technology has been widely used in bladder cancer research, opening up new ways to understand the biology of subtle bladder tumors by identifying different cell subpopulations, immune microenvironment, and single-cell technology is expected to make important changes in bladder diagnosis and treatment by identifying new biomarkers and using targeted therapies. This paper discussed the application of single-cell sequencing in bladder cancer research and clinical practice by summarizing recent advances in heterogeneous cell subpopulations, developmental patterns, and immune microenvironment and drug resistance in bladder cancer.