ObjectiveTo investigate the characteristics of pelvic floor structure and electrophysiology in female patients with stroke. MethodsFrom June to December, 2020, 21 female inpatients with stroke in Zhongda Hospital, Southeast University (stroke group) were divided into urinary incontinence (UI) group (n = 6) and non-urinary incontinence (NUI) group (n = 15), and other 20 healthy subjects were as control group. They were observed with pelvic floor ultrasonography and pelvic floor surface electromyogram. ResultsAverage electromyography, integral electromyography, root mean square, mean power frequency and median frequency decreased in UI and NUI groups compared with those of the control group (P < 0.05), but there was no significant difference between UI group and NUI group (P > 0.05). Bladder neck position, bladder neck angle, bladder neck mobility, urethral rotation angle; and anteroposterior diameter, left-right diameter and area of levator ani muscle hiatus after Valsalva's action were all not different among three groups (F < 2.484, P > 0.05). ConclusionThe activities of pelvic floor muscles decrease in female patients with stroke, without obvious changes of pelvic floor supporting structures, whatever UI.

OBJECTIVETo observe the effects of heat-sensitive moxibustion on corticotropin releasing hormone (CRH), adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) in rats with irritable bowel syndrome (IBS), and to explore the possible mechanism of heat-sensitive moxibustion on IBS.

METHODSAccording to random number table, 56 SD male rats were randomly divided into a blank group (=8), a model group (=8), a moxibustion group (=32), and a mifepristone group (RU-486 group,=8). The rats in the blank group were treated with normal feeding; the rats in the model group, RU-486 group and moxibustion group were treated with chronic non-predictable stimulation for 21 days to establish IBS model. After model establishment, the rats in the moxibustion group were treated with moxibustion at "Mingmen" (GV 4) for 40 min, once a day for 14 days; the tail temperature was recorded every 5 min; according to the change of tail temperature, the rats were divided into a heat-sensitive moxibustion group and a non-heat-sensitive moxibustion group, and 8 rats were randomly selected in the two groups. The rats in the RU-486 group were treated with gastric administration of RU-486 for 14 days, while the rats in the blank group, model group and moxibustion groups were treated with identical volume of 0.9% NaCl. The rat general condition, body mass, behavioristics, intestinal propulsive rate and visceral sensitivity were observed in each group; ELISA method was used to detect serum CRH, ACTH and CORT; optical microscope was applied to observe the morphological changes of colon.

RESULTS(1) After model establishment, rats were in rest state, fatigued, with withered hair and dim ear; the stool was dry or watery; the body mass were slowly increased; the number of crossed grid and standing frequency were significantly reduced; visceral sensitivity was increased and intestinal propulsion rate was decreased; no obvious inflammatory cell infiltration was observed under microscope. (2) After intervention, compared with the blank group, the body mass and visceral sensitivity in the RU-486 group were not significantly different (both>0.05), but the intestinal propulsion rate was decreased significantly (<0.01). Compared with the blank group, the body mass of heat-sensitive moxibustion group and non-heat-sensitive moxibustion group was lower (both<0.01), but the visceral sensitivity and intestinal propulsion rate were similar (both>0.05). Compared with the model group, the body mass and visceral sensitivity were improved in the RU-486 group (<0.05,<0.01), but the intestinal propulsion rate was similar (>0.05). The body mass, visceral sensitivity and intestinal propulsion rate of the heat-sensitive moxibustion group and the non-heat-sensitive moxibustion group were superior to those of the model group (<0.05,<0.01), and the body mass and intestinal propulsion rate of heat-sensitive moxibustion group were superior to those of non-heat-sensitive moxibustion group (both<0.05). (3) After intervention, compared with the blank group, the contents of CRH, ACTH and CORT in the model group were significantly increased (<0.05,<0.01). Compared with the model group, the contents of CRH, ACTH and CORT of the heat-sensitive moxibustion group were statistically reduced (<0.05,<0.01), and the contents of CRH and ACTH in the non-heat-sensitive moxibustion group were statistically reduced (<0.05,<0.01); the content of CRH in the RU-486 group was reduced (<0.05), but the contents of ACTH and CORT were increased (<0.05,<0.01). Compared with the non-heat-sensitive moxibustion group, the heat-sensitive moxibustion group was better in the improvement of CRH (<0.05), but there was no significant difference of ACTH and CORT between the two groups (both>0.05).

CONCLUSIONHeat-sensitive moxibustion could reduce the contents of CRH, ACTH and CORT through the HPA axis, and improve the function of gastrointestinal motility to treat IBS.

Objective To establish a quick method to analyze vinorelbine ( NVB) in plasma of beagle dogs and study its pharmacokinetics of long-circulation and thermosensitive liposome loaded vinorelbine bitartrate.Methods The plasma was treated with liquid-liquid extraction after precipitation in methanol.The analysis was perfomed on a Venusil XBP C18 column(2.1 mm ×50 mm, 3 μm) at 35℃,the mobile phase consisted of methanol and water( containing 10 mmol/L ammonium acetate, 1%acetonitrile) 80∶20 and injection volume was 10μl.The type of mass spectrum was multireactive monitoring(MRM) in a positive mode.The monitor transitions were m/z 779.4-765.4 for vinorelbine and m/z 825.4-122 for vincristine.Results The concentration range from 10 to 2500 ng/ml had a good linearity ( r=0.0994).The precision, accuracy and extraction efficiency were acceptable.The plasma samples were stable for 10 days at -20℃ and 24 h at room temperature.Pharmacokinetic study in beagle dogs showed that the main parameters for injection and liposome were Cmax(833.51 ±150.42) and (1397.95 ±443.05)ng/ml, AUC(0-t) (577.16 ±223.57) and (1059.82 ±408.27) ng/ml· h, Cl(3014.64 ±1049.17)and 1633.10 ±551.77 ml/(h· kg), respectively.Conclusion A reliable HPLC-MS/MS method for vinorelbine analysis is established and can be applied to the pharmacokinetics study of liposome.The results show that liposome has a higher AUC, Cmax and longer Cl than injection.Meanwhile, liposome has a lower irritability.

Purpose To investigate the effects of chemotactic factor CCR4 on the abi1ity of pro1iferation,ce11 cyc1e,invasion,and mi-gration of human ga11b1adder cancer ce11. Methods Western b1ot was used to detect the expression 1eve1 of CCR4 in ga11b1adder carci-noma ce11s. Ga11b1adder carcinoma ce11s was infected by means of s1ow virus,the CCR4 gene si1encing was conducted using siRNA-CCR4 interference techno1ogy. Ga11b1adder carcinoma ce11s GBC-SD were divided into three groups( GBC-SD,GBC-SD/CCR4-RNAi and GBC-SD/contro1). CCL17,a 1igand of CCR4,was used to act on these three groups of ce11s. CCK8 method was used to detect the ce11 pro1iferation abi1ity of three groups. F1ow cytometry was used to test ce11 cyc1e. Tanswe11 assay was app1ied to detect ce11 migration and invasion abi1ity. Western b1ot was performed to detect the expression of its corresponding 1igands CCL17 and CCL22 proteins. Re-sults CCR4 gene si1ence did not inf1uence ce11 cyc1e and pro1iferation of ga11b1adder ce11 GBC-SD,but can significant1y inhibit GBC-SD ce11 invasion and movement abi1ity,CCR4 gene si1ence had no inf1uence on the expression of CCL17 and CCL22 gene in tumor ce11s. Conclusion Ga11b1adder carcinoma ce11s GBC-SD express chemokine receptor CCR4,chemokine receptor CCR4 can promote the invasion and metastasis of GBC-SD ce11s.

An on-line solid phase extraction ( SPE ) coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps:A protein precipitation extraction used methanol and acetonitrile ( 50:50 , V/V ); an on-line SPE treatment to remove most matrixes in plasma;an enrichment and separation step used a C18 analytical column. S-and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin (10 mm × 2. 1 mm × 5 μm), while the chromatographic separation was achieved using a ZORBAX SB-C18 (50 mm × 2. 1 mm × 3. 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (40:60:0. 1, V/V/V, 0. 3 mL/min). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292 . 1/154 . 0 and m/z 260. 4/116. 2 were used to measure S-ammuxetine, R-ammuxetine and internal standard (propranolol). The method was linear over a concentration range from 0 . 2 to 1000 μg/L with the correlation coefficients of 0 . 9903 and 0 . 9951 . The average intra-day precision values were 1 . 2% -12 . 0% for S-ammuxetine and 0. 4%-11. 2% for R-ammuxetine, respectively. The average recoveries were 94. 2%-101. 6% for S-ammuxetine and 94. 3% -109. 4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

Objective To investigate the pathologic mechanism of radiofrequency ablation ( RFA ) combined with intravenous infusion of thermosensitive liposome encapsulated vinorelbine (TL-Vin) in treating liver tumors, and to analyze the effect of combination therapy on the long-term survival rate. Methods H22 liver adenocarcinoma tissue was subcutaneously implanted into ICR mice to establish the animal models. At the first experimental period, 40 mice were randomly and equally divided into 5 groups to receive different therapeutic scheme (using different TL-Vin concentrations). Twenty-four hours after the treatment the tumor specimens were collected, the necrotic areas were measured separately, and the optimal TL-Vin concentration was determined. At the second experimental period, 13 mice were randomly selected to receive treatment. Half an hour after the treatment the tumor tissues were collected and the TL-Vin concentration within the tumor was determined. At the third experimental period, 32 mice were randomly and equally divided into 4 groups, and 90 days after treatment the tumor growth curve was drawn. The survival rate was compared between each other of the groups. Results Compared with pure RFA group, TL-Vin + RFA significantly increased tumor coagulation extent (P<0.01). But free-VIN+RFA had similar tumor necrotic extent as that produced by RFA alone (P>0.05). Tumor coagulation area in TL-Vin + RFA group was bigger than that in free-VIN + RFA group at the concentration of 10 mg/kg [(341.8 ± 65.4)mm2 vs (225.3 ± 25.4)mm2, P < 0.01]. In TL-Vin group the coagulation margin was clear. The mean intratumoral Vinorelbine accumulation in TL-Vin + RFA group was 10 folds of that in free-Vin group [(1 156.5 ± 158.3)ng/ml vs (194.5 ± 52.3)ng/ml, P = 0.005]. TL-Vin +RFA had better survival result than that of RFA alone (37.6 ± 20.1 days vs. 23.4 ± 5.0 days, P=0.015), as well as than that of free-Vin + RFA [(37.6 ± 20.1)days vs (23.3 ± 1.2)days, P = 0.016]. Conclusion Thermosensitive liposomal chemotherapies (Vinorelbine) can be selectively delivered at the edge of RFA coagulation area and thus effectively increase RFA-induced tumor coagulation and prolong the end-point survival in experimental mice.

ObjectiveTo evaluate the influence of electrotome on permanent and temporary cardiac pacemaker in laparoscopic cholecystectomy ( LC),and the application of cardiac pacemaker to the cases of cholecystolithiasis combined with bradyarrhythmia.MethodsClinical data of 215 patients with permanent or temporary cardiac pacemaker who underwent were studied for the preoperative and postoperative variation of pacemaker function,and for the influence of electricity coagulation during the operation on cardiac pacemaker function.ResultsLC was successfully completed in all 215 patients.The function of cardiac pacemaker was not obviously interfered during the operation,and the parameters of cardiac pacemaker did not remarkably change after the operation.ConclusionCardiac pacemaker is slightly interfered when electrotome and electrocoagulation were used in LC; LC is feasible and safe for patients with bradyarrhythmia by placement of cardiac pacemaker.

Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia (AA), high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ± 0. 29, 2. 26 ± 0. 37 and 1.58±0. 33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively (P < 0.01). The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0. 11,respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.

AIM To analyze the toxicity and inhibitory mechanism of extracts from cultured cells (F 4 cell line) of Taxus chinensis on cancer cell lines SMMC 7721 and HEp 2. METHODS MTT assay for cell viability and flow cytometry for cell cycle analysis. RESULTS IC 50 of SMMC 7721 and HEp 2 were 0 161 4 g DCW?L -1 and 0 275 6 g DCW?L -1 respectively,tumor cells in G 2～M stage all increased with higher concentration and longer incubation of extracts from Taxus chinensis cells. CONCLUSION Extracts from cultured cells of Taxus chinensis could have cytotoxic effect on SMMC 7721 and HEp 2 and could induce apoptosis of both two cancer cells.

Src is a non-receptor protein tyrosine kinase activated by a number of extracellular signal moleculars. It is recruited to peripheral sites through myristoylation and the SH3 domain. Src initiates intracellular signal trandsduction pathways that influence cell adhesion, migration, growth, differentiation and survival though catalytic domain. Src is normally maintained in an inactive conformation because of carboxy terminal Src kinase, but can be activated transiently during cellular events such as mitosis or constitutively by abnormal events such as mutation and some cancers. In additions, c-Src protein is found to be highly activated and the Src gene is frequently over-expressed in many cancers. These findings suggest that the relationship between c-Src activation/over-expression and cancer progression appears to be significant.