OBJECTIVE:To establish a method for the content determination of total flavonoids in Morus alba. METHODS:UV-visible spectrophometry was performed with Al(NO2)3-NaNO2-NaOH color-test at the wavelength of 510 nm with the reference of rutin. RESULTS:The linear range of rutin was 0.031 2-0.156 mg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 95.7%-101.0%(RSD=2.1%,n=6). CONCLUSIONS:The method is simple,sta-ble and reproducible,and can be used for the content determination of total flavonoids in M. alba.

Objective To evaluate the effect of mitochondrial permeability transition pore (mPTP) in lipid emulsion-induced inversion of bupivacaine myocardiotoxicity in rats.Methods H9c2 cells were inoculated in 6-well plates at a density of 105 cells/ml, and were randomly divided into 4 groups (6 wells in each group, 2 ml/well) using a random number table: control group (group C) , bupivacaine group (group B) , lipid emusion + bupivacaine group (group LB) , and lipid emusion + bupivacaine + atractyloside group (group LBA).Phosphate buffer solution 100 μl was added to the culture medium in group C.In group B, bupivacaine was added to the culture medium with the final concentration of 1 mmol/L.In group LB, lipid emusion and bupivacaine were added to the culture medium with the final concentrations of 1％ and 1 mmol/L, respectively.In group LBA, lipid emusion, bupivacaine and atractyloside (an mPTP opener) were added to the culture medium with the final concentrations of 1％, 1 mmol/L and 30 μmol/L, respectively.All the cells were incubated for 24 h.After the end of incubation, the expression of Bcl-2, Bax, phosphorylated Bad (p-Bad) , caspase-3, activated caspase-3, caspase-9,activated caspase-9 and cytochrome c (Cyt c) was detected using Western blot.The expression of Bcl-2 mRNA, Bax mRNA, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was detected using real-time reverse transcriptase polymerase chain reaction.The ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were calculated.Results Compared with group C,the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/ Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group B (P＜0.05) , and no significant change was found in the parameters mentioned above in group LB (P＞0.05).Compared with group B, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly decreased, the expression of p-Bad was up-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was down-regulated in LB and LBA groups (P＜ 0.05).Compared with group LB, the ratios of Bax/Bcl-2, activated caspase-3/caspase-3, activated caspase-9/caspase-9, and Bax mRNA/Bcl-2 mRNA were significantly increased, the expression of p-Bad was down-regulated, and the expression of Cyt c, Bad mRNA, caspase-9 mRNA and Cyt c mRNA was up-regulated in group LBA (P ＜ 0.05).Conclusion The mechanism underlying lipid emulsioninduced inversion of bupivacaine myocardiotoxicity is related to inhibited mPTP opening in rats.

Objective To observe the safety and effectiveness of inductive chemotheray with lobaplatin plus 5-Fu (LF regimen) and concurrent chemoradiotherapy with lobaplatin for local-regionally advanced nasopharyngeal carcinoma (NPC) patients,and investigate the appropriate lobaplatin dose for the concurrent chemoradiotherapy.Methods Newly diagnosed local-regionally advanced NPC patients signed informed consent.The inductive chemotherapy was lobaplatin 30 mg/m2 + 5-Fu 4 g/m2 civ 120 h for 2 cycles every 21 days,then concurrent lobaplatin chemoradiotherapy was conducted.The initial lobaplatin dose for concurrent chemoradiotherapy was 50 mg/m2 with at least 3 cases in every dose level.If 2 of 3 patients presented dose-limiting toxicity (DLT),5 mg/m2 dose decreased for the next level until maximal tolerant dose (MTD) reached.The tumor response was evaluated after inductive chemotherapy,at the end of the chemoradiotherapy,3 months after chemoradiotherapy and 6 months after chemoradiotherapy.Results From Dec 2011 to Apr 2012,11 patients were enrolled in this study.After 2 courses of inductive chemoradiotherapy,CR,PR and SD were observed in 1,8 and 2 patients,respectively.At the end of the chemoradiotherapy and 3 months after chemoradiotherapy,CR and PR were observed in 10 and 1 patients,respectively.Six months after the chemoradiotherapy,all patients were CR.For the patients(3 in each arm) received 50 mg/m2 or 45 mg/m2 lobapaltin concurrent chemoradiotherapy,2 patients in each arm presented DLT.For the 5 patients received 40 mg/m2 lobapaltin concurrent chemoradiotherapy,no patients presented DLT.40 mg/m2 was suggested as the MTD.Inhibition of platelet was the major DLT.Conclusion Inductive chemotherapy with LF regimen and concurrent chemoradiotherapy with lobaplatin is safe and effective for local-regionally advanced NPC patients and the MTD of lobaplatin for the concurrent chemoradiotherapy is 40 mg/m2.Further clinical trial with large sample is expected.

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Objective To design a laser acupuncture instrument with dual wavelength and multi-channel,under the guidance of traditional Chinese medicine acupuncture theory and according to low-level laser induced effect in bio-tissues and compatibility characteristics of acupuncture points.Methods Using SCM and semiconductor laser,hardware circuitry was designed to achieve the basic functions of the instrument,and the corresponding PC software was developed.Results The laser acupuncture treatment for several acupuncture points could be carried out at the same time with the same or different laser parameters (wavelength,power density,continuous/pulse),and each channel could output 635 nm and 808 nm laser,with continuous adjustable power.Conclusions The dual wavelength multi-channel laser acupuncture instrument can meet the needs of compatibility during clinical acupuncture treatment and is easy to operate.

Aim To investigate the effect of gelsemium alkaloids on chloride channels and cell volume in he-patic carcinoma cells. Methods The time-lapse live cell imaging and whole-cell patch clamp techniques were used respectively to detect the volume changes and currents induced by gelsemium alkaloids in HepG2 cells. Results It was found that the cell volume was decreased by (12. 48 ± 2. 2) % (P<0. 01) when ex-posed to gelsemium alkaloids for 50 min and this phe-nomenon could be inhibited by the chloride channel blocker tamoxifen. It was shown by whole-cell patch clamping that a chloride current could be evoked by extracellular application of gelsemium alkaloids ( 2μmol·L-1 ) . The current was outward-rectified with-out obvious voltage- and time-dependent inactivation. The reversal potential of the current was ( -3. 21 ± 0. 67) mV ,which was close to the equilibrium poten-tial of chloride. The extracellular application of the chloride blockers, tamoxifen and 5-notro-2-(3-phenyl-propylamino)benzoic acid (NPPB), and 47% hyper-tonic solution inhibited the current significantly ( P <0. 01 ) . Conclusion Gelsemium alkaloids could acti-vate chloride channels and induce a volume decrease ( named apoptotic volume decrease, AVD) , and these effect could be inhibited by chloride channel blockers. The results suggest that the chloride channel can be one of the targets of gelsemium alkaloids in their anti-cancer action.

Objective To compare the outcomes of percutaneous Kirschner wire assisted reduction with minimally invasive plate osteosynthesis and open reduction with locking plate in treatment of Robinson type 2B midshaft clavicular fractures.Methods A retrospective case control study was conducted to analyze the clinical data of 96 patients with Robinson type 2B midshaft clavicular fracture from June 2009 to October 2016.There were 70 males and 26 females with an average age of 34.5 years.The patients were divided into two groups according to the different surgical methods:49 patients were treated with percutaneous Kirschner wire assisted reduction and minimally invasive locking plate osteosynthesis (minimally invasive group);47 patients were treated with open reduction with locking plate internal fixation (open plating group).According to the Robinson fracture typing,there were 28 patients with type 2B1 and 21 with type 2B2 in the minimally invasive group;there were 23 patients with type 2B1 and 24 with type 2B2 in open plating group.The length of incision,operation time,visual analog scale (VAS) 2 days after surgery,length of hospital stay,time of fracture healing,Constant score results of postoperative 6 months,1 year,and at the last follow-up,feedbacks on incision and functional satisfaction,and complications were compared between the two groups.Results The average follow-up was (19.6 ± 5.5)months in minimally invasive group and (20.3 ± 6.2)months in open plating group.The incision lengths of the minimally invasive group and the open plating group were (4.8 ±0.7)cm and (8.3 ± 1.6) cm,respectively.The minimally invasive group had significantly longer operation time [(75.5 ±21.6)minutes] compared with open plating group [(60.2 ± 19.1)minutes] (P ＜0.0l).In the minimally invasive group and open plating group,the VAS 2 days after surgery was (2.9 ± 1.5) points and (3.9 ± 1.7) points;the hospitalization time was (7.1 ± 2.6) days and (9.5 ± 2.9) days;the fracture healing time was (11.7 ± 2.2) weeks and (13.4 ± 2.9) weeks;patients' satisfaction with the appearance of the incision was 84％ and 60％,respectively.There were no significant differences in the Constant score and functional satisfaction between the two groups at postoperative 6 months,l year,and the last follow-up (P ＞ 0.05).The incidence of postoperative incisional hypertrophic scar was 8％ and 30％,and the incidence of local skin numbness or sensory loss was 10％ and 32％ in minimally invasive group and open plating group,respectively (P ＜ 0.0l).No significant differences in nonunion,internal fixation failure,incision infection,subcutaneous prominent plate or revision rate were found between the two groups (P ＞ 0.05).Conclusion Percutaneous Kirschner wire assisted reduction with minimally invasive locking plate and open reduction with locking plate internal fixation can both achieve good clinical outcomes in the treatment of Robinson 2B midshaft clavicular fracture;but the former shows more advantages in terms of incision length,incision appearance,operation time,pain 2 days after surgery,fracture healing time,and local skin sensory disturbances.

OBJECTIVETo elucidate the role of transforming growth factor-beta1(TGF-β1) in epithelial-mesenchymal transition of mesothelial cells and peritoneal metastasis of gastric cancer.

METHODSHMrSV5 cells, a human peritoneal mesothelial cell line, were incubated with TGF-β1, and their morphological changes were observed by phase contrast microscopy. Expressions of α-smooth muscle actin (α-SMA), vimentin, cytokeratin, E-cadherin, phosphorylated-Smad2 and Smad2 were examined by Western blotting. After fibroblastic-like mesothelial cells were co-incubate with HSC-39 cells(gastric cancer cell line), the adhesion and invasion potential of HSC-39 were evaluated by adhesion and invasion assay in vitro.

RESULTSFew mesothelial cells converted to spindle fibroblast-like morphology for 24 h, and remarkable phenotypic changes were observed at 72 h of TGF-β1 activation. TGF-β1 could induce α-SMA and vimentin expression, and down-regulate cytokeratin and E-cadherin expression in mesothelial cells (P<0.05). TGF-β1 induced phosphorylation of Smad2 within 15 min of stimulation, reached a maximum at 30 min after treatment and remained high level during the experiment without affecting total Smad2 expression(P>0.05). The percentage of HSC-39 gastric cancer cells adhered were significantly increased as compared to the control. When the mesothelial cells were treated by TGF-β1 for 72 h, the increased adhesion percentage was(146±17)%(P<0.05). After fibroblastic-like mesothelial cells co-incubated with HSC-39 cells for 48 h, more cancer cells [(61.1±11.4) cells/view field] invaded the coated membrane as compared to the control group [(31.9±8.1) cells/view field] (P<0.05).

CONCLUSIONTGF-β1 can induce the transition of mesothelial cells into myofibroblasts and Smad2 signal pathway may play a role in this transition, which is associated with increased adhesion and invasiveness of gastric cancer cells, and provides favorable environment for the dissemination of gastric cancer.

Cadherins ; Cell Line, Tumor ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Epithelium ; Fibroblasts ; Humans ; Peritoneal Neoplasms ; Signal Transduction ; Smad2 Protein ; Stomach Neoplasms ; Transforming Growth Factor beta1 ; Vimentin

Objective:To analyze the long-term efficacy and safety of re-irradiation for recurrent glioma.Methods:The data of 52 patients with recurrent gliomas were collected from 2009 to 2019. The median planned targetvolume (PTV) was 73.5 cm 3(49.9-102.7 cm 3) and the median dose was 45.0 Gy (43.0-48.8 Gy). Kaplan-Meier method was used for survival assessment, log-rank test for difference assessment, and Cox’s regression model for multivariate prognostic analysis. Results:The median follow-up time was 32.6 months. The median overall survival (OS) and progression-free survival (PFS) time were 16.1 months (95% CI, 4.1-28.1) and 8.0 months (95% CI, 4.0-12.0). The 1-, 2-and 3-year survival rates were 67%, 43% and 29%, respectively. The 6-month, 1-year and 2-year PFS rates were 67%, 40%, 26%, respectively. Multivariate analysis showed that KPS score and recurrence time significantly affected the OS ( P=0.012, P=0.001). KPS score and time interval between two radiotherapies significantly impacted the PFS ( P=0.003, P=0.018). Stratified analysis showed that KPS score was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅱ initial pathology and reoperation after recurrence ( P<0.001, P=0.012); clinical manifestation was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅲ and Ⅳ initial pathology ( P=0.006, P=0.044). The overall incidence of adverse reactions was 30.8%. Grade 1 adverse reactions accounted for 25.0%, and 5.8% for grade 2. Conclusions:Re-irradiation for recurrent glioma yields good long-term clinical efficacy and tolerable adverse reactions.

Objective:To distinguish the differences between Olibanum and Myrrha by using modern analytical methods such as Differential Scanning Calorimeter (DSC) and Fourier Transform InfraRed (FT-IR). Methods:By collecting Olibanum and Myrrha in different growing areas and different processed prosecures, this paper to analizes the influence of temperature increase and its speed , as well as the particle size on the DSC experiment. The DSC method was used to perform a differential thermal map of Olibanum and Myrrha scanning and analysis; FT-IR was used to scan and analyze 20 batches of Olibanum and Myrrha. Results:TThe results of DSC analysis showed that the DSC experimental condition ranged between 30-600 ℃; the speed of temperature increase was 30 ℃/min; the particle size was 100 mesh. The DSC spectra of of Olibanum and Myrrha were significantly different. Only the processed products of frankincense had endothermic peak near 297 ℃, and there was no characteristic peak in this temperature range. Their exothermic peaks are close at 326 ℃, but their enthalpy values are quite different. The position of endothermic peak near 100 ℃ is close to the size of peak shape. FT-IR test showed that the absorption peaks of Olibanum and Myrrha at wave numbers 2 925, 1 710, 1 454, 1 371, 1 242, 1 029 cm -1 appeared, and the positions of strong peaks were also similar. The intensity of the characteristic peak of Myrrha wave number 1 029 cm -1 is greater than that of Olibanum. Conclusion:The DSC spectra of Olibanum and Myrrha are significantly different, and the difference between the two FT-IR spectra is small. Differential scanning Calorimetry is an effective, fast, and simple way to identify resinous Chinese medicinal materials, and is worthy of further popularization.