OBJECTIVE:To establish a method for the simultaneous determination of gallic acid,rhein and emodin in Com-pound xiaozhi suppository. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of 0.5% phosphor-ic acid-Acetonitrile solution(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 273 nm for gallic acid and 254 nm for rhein and emodin,the column temperature was 28 ℃,and the injection volume was 20 μl. RESULTS:The linear range was 0.296 4-14.82 μg/ml for gallic acid(r=0.999 6),0.215 0-10.75 μg/ml for rhein(r=0.999 9)and 0.307 2-15.36 μg/ml for emo-din(r=0.999 9);RSDs of precision,stability and reroducibility tests were lower than 3.0%;recoveries were 95.1%-97.2%(RSD=0.64%,n=6),95.4%-97.2%(RSD=0.42%,n=6) and 96.5%-99.4%(RSD=1.10%,n=6),respectively. CONCLU-SIONS:The method is simple and accurate,and can be used for the contents determination of gallic acid,rhein and emodin in Compound xiaozhi suppository.

OBJECTIVE:To establish a RP-HPLC method to determine concentration of cefixime in human serum.ME_ THODS:The assay was conducted on Agilent ZORBAX?SB C 18 column with methanal-water-PBS(25∶15∶60,V/V/V)as mobile phase.The detection wavelength was270nm,and the acetanilide was taken as internal standard.RESULTS:The lin?earity range was0.11～5.28mg/L with a regressive equation of Y-0.07534X-0.13186,r=0.9995.The recovery was high.CONCLUSION:This method is accurate,rapid and simple and can be used for determining the concentration of cefixime in human serum.

To observe the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, in inhibiting the growth of transplanted A549 tumors in nude mice and the expressions of p-Akt and HIF-1alpha in A549 tumors.

Objective: To develop an HPLC method for the simultaneous determination of paeoniflorin , liquiritin, baicalin and costunolide in Fuke Yangkun pills , and optimize the extraction technology by response surface methodology ( RSM).Methods: The separation of targeted compounds were performed on a Kromasil C 18 column (250 mm ×4.6 mm, 5 μm).The mobile phase consisted of acetonitrile (A) and 0.2%phosphoric acid (B) with gradient elution at a flow rate of 1.0 ml min-1.The detection wavelength was set at 230, 276, 280 and 225 nm, respectively.The column temperature was 28℃.Using the contents of the four components as the indices, the extraction process was optimized by a response surface method with methanol concentration , solid-liquid ratio and ex-traction time as the influencing factors .Results: The linear range of paeoniflorin , liquiritin, baicalin and costunolide was 1.616-161.600, 0.432-43.180, 2.045-204.500 and 0.518-51.840 μg ml-1, respectively.The average recovery (n=9) was 98.3%, 99.6%, 97.9%and 98.1%, respectively.The optimum conditions of extraction process were as follows:the methanol concentration was 64%, the solid-liquid ratio was 1 ∶51, and the extraction time was 25 min.Conclusion: The response surface methodology is convenient and highly predictive in optimizing the extraction process of the four effective components in Fuke Yangkun pills .The devel-oped content determination method is simple and accurate , which can be used for the quality control of Fuke Yangkun pills .

Background: Tumor markers are widely used in clinical practice and have become important indicators in assessing cancer progress. There is increasing concern that chemotherapy combined with traditional Chinese medicine has effects in decreasing the level of tumor markers. Objective: To investigate the effects of chemotherapy combined with Kangliu Zengxiao Decoction (KLZX), a compound Chinese herbal drug, on tumor markers carbohydrate antigen 50 (CA 50), cytokeratin 19 fragment (CYFRA21-1) and carcinoembryonic antigen (CEA) in patients with advanced non-small-cell lung cancer (NSCLC) and to explore the relationships between clinical efficacy and tumor markers. Design, setting, participants and interventions: Patients were included from Punan Hospital of Shanghai Pudong New District and Longhua Hospital between October 2008 and December 2009. Seventy-four subjects with advanced NSCLC were randomly assigned into treatment group (n=37) and control group (n=37). Patients in the control group were treated with chemotherapy alone while patients in the treatment group were treated with chemotherapy combined with KLZX. Chemotherapy of NP (vinorelbine + cisplatin) was given for two cycles and patients in the treatment group were administered with KLZX during chemotherapy. Main outcome measures: Levels of CA50, CYFRA21-1 and CEA before and after treatment were evaluated and the relationship between changes in levels of tumor makers and tumor size, clinical symptoms and living condition score (Karnofsky score) was analyzed. Results: No patients achieved a complete remission. The disease control rates (complete remission (CR)+partial remission (PR)+no change (NC)) were 89.20% (33/37) and 70.30% (26/37) in the treatment and control group respectively (P<0.05). The levels of CA50, CYFRA21-1 and CEA were clearly decreased in the treatment group after treatment (P<0.05) while also decreased in the patients without progression of disease. There were no obvious changes of CA50, CYFRA21-1 and CEA in the control group, and there was even a trend of increase. Furthermore, the improvement rates of clinical syndrome were 51% (19/37) vs 11% (4/37) (P<0.05) in the treatment group and control group respectively. The total response rates of quality of life were 91.89% (34/37) vs 56.76% (21/37) (P<0.01) in the treatment and control group respectively. Conclusion: Combined chemotherapy with KLZX in treating advanced NSCLC can acquire better stabilizing tumor foci, decrease levels of tumor markers and improve the clinical symptoms and Karnofsky score.

Objective To evaluate effects of alimentary reconstruction procedures (integral continual jejunal interposition, Billroth Ⅱ and isolated jejunal interposition) after subtotal gastrectomy on postoperative plasma gastrin, motilin and cholecystokinin. Methods Twenty-four dogs were divided into 3 groups undergoing distal subtotal gastrectomy and three different digestive tract reconstruction (integral continual jejunal interposition, Billroth Ⅱ and isolated jejunal interposition). The concentration of plasma gastrin, motilin and cholecystokinin were detected by enzyme-linked immunosorbent assay before and after operation. Results Two months after operation, plasma gastrin level of the integral continual jejunal interposition group (2. 2 ±0. 7 ) ng/L, ( 3.9 ± 0. 8 ) ng/L was significantly lower than that of preoperative both in fasting and postprandial state (3.8 ± 1.0) ng/L, (5.3 ± 1.6) ng/L, all P ＜0.05, but was significantly higher than other two groups in postprandial state (2. 7 ± 1.0) ng/L, (3.6 ±0. 6) ng/L, P ＜0. 05. Two months after operation, plasma motilin concentration of integral continual jejunal interposition group (577 ±204) ng/L, (1003 ± 209) ng/L were significantly higher than that of preoperative both in fasting and postprandial (429 ± 128) ng/L, (854 ± 218 ) ng/L, P ＜ 0. 05. The postoperative plasma motilin of integral continual jejunal interposition group ( 1003 ± 209 ) ng/L was significantly higher than other two groups in postprandial state (840 ±205) ng/L, (986 ± 189) ng/L, P ＜0. 05. Two months after operation,plasma cholecystokinin concentration of integral continual jejunal interposition group ( 19.6 ± 2.0 ) ng/Lwere significantly higher than that of preoperative both in postprandial ( 19.0 ± 2. 0) ng/L, P ＜ 0. 05. The postoperative plasma cholecystokinin of integral continual jejunal interposition group ( 19. 6 ± 2. 0) ng/L was significantly lower than other two groups (22.2 ± 2. 1 ) ng/L, (20. 1 ± 2. 5 ) ng/L, P ＜ 0. 05. Conclusion Integral continual jejunal interposition after distal gastrectomy maintains the postoperative plasma motilin and gastrin in a relatively higher level and decreases the concentration of plasma cholecystokinin.

To explore the dual regulatory effects of Shuanghuang Shengbai Granule, a compound traditional Chinese herbal medicine, on cell cycle in Lewis-bearing mice with chemotherapy-induced myelosuppression.

Objective:To evaluate the role of ferroptosis in the dorsal root gangions in neuropathic pain (NP) in rats.Methods:Thirty-two healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 180-220 g, were randomized into 4 groups ( n=8 each) using a random number table method: sham operation group (Sham group), NP group, NP+ solvent control group (NP+ Veh group), and NP+ liproxstatin-1 (Lip-1) group (NP+ Lip group). NP was induced by chronic constrictive injury (CCI) to sciatic nerve in anesthetized animals.In NP+ Lip group, liproxstatin-1 (diluted to 10 μg/μl in DMSO) 30 μl was intrathecally injected for 3 consecutive days after surgery.NP+ Veh group received intrathecal injection of DMSO 30 μl for 3 consecutive days after surgery.Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 3 days before surgery and on days 1, 3, 5, 7 and 10 after surgery.Rats were sacrificed after the end of pain threshold measurement on day 10 after surgery, and DRGs of the lumbar segment (L 3-5) on the left side were removed for determination of the levels of iron ion, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD), expression of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot), and expression of ACSL4 in each nerve cells of DRGs (by immunofluorescence) and for microscopic examination of the ultrastructure of mitochondria in DRGs (by transmission electron microscopy). Results:Compared with Sham group, MWT was significantly decreased and TWL was shortened at T 2-6, levels of iron ions, ROS and MDA in DGRs were increased, activities of SOD and GSH-Px were decreased, ACSL4 expression was up-regulated, GPX4 expression was down-regulated, and ACSL4 expression in astrocytes and Schwann cells of DRGs was up-regulated in NP group ( P<0.05). Compared with NP group, MWT was significantly increased and TWL was prolonged at T 3-6, levels of iron ions, ROS and MDA in DGRs were decreased, activities of SOD and GSH-Px were increased, ACSL4 expression was down-regulated, GPX4 expression was up-regulated, and ACSL4 expression in astrocytes and Schwann cells of DRGs was down-regulated ( P<0.05), and no significant change was found in the parameters mentioned above in NP+ Veh group ( P>0.05). The results of electron microscopy showed that collapsed mitochondrial cristae and membrane rupture were found in astrocytes and Schwann cells of DRGs in NP group, and the number of collapsed mitochondrial cristae and membrane rupture was significantly decreased in NP + Lip group when compared with NP group. Conclusions:The ferroptosis in DRGs is involved in NP in rats.

Objective To investigate the effects of bufalin in reversing hepatocyte growth factor ( HGF)?induced resistance to afatinib in H1975 lung cancer cells, and explore its possible mechanism. Methods The afatinib?resistant H1975 lung cancer cells ( H1975AR) were induced by exogenous HGF and transfected with recombinant adenoviral vector Ad?HGF?GFP. The cytostatic effects of bufalin, afatinib and bufalin plus afatinib on H1975AR cells were evaluated by MTT assay. The impact of combined therapy with bufalin and afatinib on invasion of H1975AR cells was determined by transwell migration assay. The concentrations of HGF in the culture supernatants of H1975/Vec and H1975/HGF cells were determined by ELISA. The expression of EGFR, cMET and EMT signal pathway?related proteins in H1975AR cells treated with bufalin, afatinib and bufalin plus afatinib were detected by Western blot. Results The results of MTT assay showed that afatinib did not inhibit the growth of H1975 cells, but after 72 h of the combined treatment with bufalin and afatinib and in the presence of HGF, the growth rate of H1975 cells was (38.67±8.76)%, significantly lower than the growth rate of (63.45±12.65)% in the H1975 cells treated with HGF alone (P<0.05) . The results of transwell migration assay showed that in the presence of HGF, afatinib plus bufalin combination therapy markedly decreased the number of invaded H1975 cells through the Matrigel chamber (48.98±11.43), significantly lower than the 118.92±37.29 of afatinib?treated or the 88.84±19.53 of bufalin?treated cells (P<0.05 for all). The result of ELISA showed that H1975/HGF cells secreted high levels of HGF, and afatinib and bufalin had no effect on the HGF secretion in H1975/HGF cells. The results of Western blot analysis showed that the expression of p?EGFR, p?cMet, p?AKT, p?ERK, vimentin and snail in H1975AR cells treated with bufalin puls afatinb was down?regulated markedly, and the expression of E?cadherin was up?regulated markedly. Conclusions Combination of bufalin and afatinib strongly inhibits the growth of H1975AR lung cancer cells and decreases their invasion ability. The possible mechanism of combined treatment with bufalin and afatinib may be related to the blocking of cMet/PI3K/AKT and cMet/MAPK/ERK pathways and inhibiting of epithelial?mesenchymal transition.

Objective To investigate the effects of bufalin in reversing hepatocyte growth factor ( HGF)?induced resistance to afatinib in H1975 lung cancer cells, and explore its possible mechanism. Methods The afatinib?resistant H1975 lung cancer cells ( H1975AR) were induced by exogenous HGF and transfected with recombinant adenoviral vector Ad?HGF?GFP. The cytostatic effects of bufalin, afatinib and bufalin plus afatinib on H1975AR cells were evaluated by MTT assay. The impact of combined therapy with bufalin and afatinib on invasion of H1975AR cells was determined by transwell migration assay. The concentrations of HGF in the culture supernatants of H1975/Vec and H1975/HGF cells were determined by ELISA. The expression of EGFR, cMET and EMT signal pathway?related proteins in H1975AR cells treated with bufalin, afatinib and bufalin plus afatinib were detected by Western blot. Results The results of MTT assay showed that afatinib did not inhibit the growth of H1975 cells, but after 72 h of the combined treatment with bufalin and afatinib and in the presence of HGF, the growth rate of H1975 cells was (38.67±8.76)%, significantly lower than the growth rate of (63.45±12.65)% in the H1975 cells treated with HGF alone (P<0.05) . The results of transwell migration assay showed that in the presence of HGF, afatinib plus bufalin combination therapy markedly decreased the number of invaded H1975 cells through the Matrigel chamber (48.98±11.43), significantly lower than the 118.92±37.29 of afatinib?treated or the 88.84±19.53 of bufalin?treated cells (P<0.05 for all). The result of ELISA showed that H1975/HGF cells secreted high levels of HGF, and afatinib and bufalin had no effect on the HGF secretion in H1975/HGF cells. The results of Western blot analysis showed that the expression of p?EGFR, p?cMet, p?AKT, p?ERK, vimentin and snail in H1975AR cells treated with bufalin puls afatinb was down?regulated markedly, and the expression of E?cadherin was up?regulated markedly. Conclusions Combination of bufalin and afatinib strongly inhibits the growth of H1975AR lung cancer cells and decreases their invasion ability. The possible mechanism of combined treatment with bufalin and afatinib may be related to the blocking of cMet/PI3K/AKT and cMet/MAPK/ERK pathways and inhibiting of epithelial?mesenchymal transition.