To explore the feasibility of chemical and biological method in evaluation of the in vitro dissolution rate of Liuwei Wuling tablet (LWT), this experiment investigated the inhibitory effect of LWT dissolving solutions on LX-2 hepatic stellate cells in 0.1% SDS dissolution medium in different dissolving periods. From these results, the cumulative dissolution rate of LWT was obtained based on the cell inhibitory rate. The dissolution rates of deoxyschizandrin, phillyrin, and Specnuezhenide were determined by HPLC method. A novel approach of self-defined weighting coefficient had been created to establish the integrated dissolution rate model. Then f2 similar factor method was used to evaluate the relevance of these two methods. The results showed that f2 values for deoxyschizandrin, phillyrin, Specnuezhenide, and the integrated dissolution were 61, 43, 61 and 75 respectively, indicating that the dissolution of multi-component integration could fully reflect the biological potency of the whole recipe. The dissolution evaluation method for multicomponent integration based on biological activity is expected to be one of the effective means for in vitro dissolution test of LWT.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Kinetics ; Quality Control ; Solubility ; Tablets ; chemistry

To investigate the absorption kinetics of Cu B-SDC/PLC-MMs in rat different intestinal segments and compared with the absorption of Cu B suspension. The in vitro everted gut sacs model was established to study the absorption characteristics of Cu B-SDC/ PLC-MMs in rat duodenum, jejunum, ileum and colon, and the content of cucurbitacin B was detected by HPLC method, and the effects of concentrations on intestinal absorption were evaluated as well. The results showed that the absorption of Cu B-SDC/PLC-MMs was linearity at different intestine segment and different concentrations (R2 > 0.9), which was consistent with zero order rate process. The Ka of different intestine segments showed a concentration-dependent increasing along with the raised concentration of Cu B-SDC/ PLC-MMs, indicating that it was likely to be a mechanism of passive absorption. The best absorption site of Cu B-SDC/PLC-MMs was ileum, and its absorptions in different intestinal segments were superior to cucurbitacin B suspension. SDC/PLC-MMs could significantly enhance the intestinal absorption of cucurbitacin B, and the study of intestinal absorption kinetics of Cu B-SDC/PLC-MMs had gave a support to its further reasonable solidfication.
Animals ; Deoxycholic Acid ; administration & dosage ; Female ; Intestinal Absorption ; Kinetics ; Male ; Micelles ; Nanoparticles ; Phospholipids ; administration & dosage ; Rats ; Rats, Wistar ; Triterpenes ; administration & dosage ; pharmacokinetics

To study the influence of stir-baked with sand on active ingredients, diarrhea and hepatoprotection of Herpetospermum caudigerum, the contents of herperione and herpetin in H. caudigerum before and after stir-baking with sand were analyzed by HPLC. The effect of stir-baked with sand on diarrhea of H. caudigerum TL was evaluated using the mean stool rate (MSR) and mean diarrheal index ( MDI) and the influence of stir-baked with sand on hepatoprotective effect of H. caudigerum TL was examined using a mouse model of CCl4-induced liver injury based on the analysis of serum ALT and AST activities. The results of HPLC analysis showed the content of herperione in H. caudigerum after stir-baking with sand decreased by 40.9% (P < 0.01) and the content of herpetin had no change. Pharmacodynamic results showed that the MSR and MDI of high-dose and middle-dose group of H. caudigerum TL after stir-baking with sand were significantly lower than that of high-dose and middle-dose group of H. caudigerum TL without stir-baking with sand; The high-dose and middle-dose of H. caudigerum TL with/without stir-baking with sand significantly alleviated liver injury as indicated by the decreased levels of serum ALT and AST, but the ALT and AST levels of high-dose and middle-dose group of H. caudigerum TL after stir-baking with sand were higher than that of H. caudigerum TL without stir-baking with sand. The results revealed that the stir-baking with sand could effectively relieve diarrhea effect of H. caudigerum TL, while it also reduces the hepatoprotection of H. caudigerum TL.
Animals ; Chromatography, High Pressure Liquid ; Cooking ; Cucurbitaceae ; chemistry ; Diarrhea ; chemically induced ; Female ; Liver ; drug effects ; Male ; Mice ; Protective Agents ; pharmacology

Objective To investigate the effects of glutamine (Gln) on proliferation and survival of small cell lung cancer H446 cells, and further to explore the potential mechanism. Methods The proliferation of H446 cells was detected at different time points (0, 24, 48, 72 and 96 h) by CCK-8 assay in Gln (+) group and Gln (-) group, and an optimal time was selected. Under the optimal time, Annexin V-FITC/PI staining, CellTiter-Glo? assay kit and flow cytometer were used to detect cell survival, cellular adenosine triphosphate (ATP) and reactive oxygen species (ROS) levels. Gln (-) group was used as the control group, under the condition of Gln deficiency, cellular ATP, cell proliferation and survival were detected after adding oxaloacetic acid (OAA) or dimethyl-α-ketoglutarate (DM-αKG). Gln (-) group was used as the control group, cellular ROS, cell proliferation, colony and survival were detected after treated with ROS scavenger N- acetyl cysteine (NAC). With different concentrations (0, 2, 5, 10 μmol/L) of glutaminase inhibitor BPTES, the optimal concentration was selected through the colony assay. The cellular ATP and ROS levels and cell proliferation were detected under the optimal concentration. H446 cells were treated with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl) ethyl sulfide (BPTES), ROS inducer hydrogen peroxide (H2O2) or the combination of them, and cell survival ratio was compared between two groups. Results The proliferation levels of H446 cells at 24, 48, which were decreased most significantly in 72 h in Gln (-) group. When 72 h was used as the optimal time, the cell survival ratio and ATP level were decreased, and the ROS level was increased, in Gln (-) group compared with those of Gln (+) group (P<0.05). There was a higher survival ratio in H446 cells in Gln (-)+OAA group and Gln (-)+DM-αKG group than that of Gln (-) group (P<0.05), but there were no significant differences in cell proliferation and ATP levels between Gln (-) group, Gln (-)+OAA group and Gln (-)+DM-αKG group. The ROS level was reduced, the cell proliferation, colony level and survival ratio were increased in Gln (-)+NAC group compared with those of Gln (-) group (P<0.05). Cloning assay showed that 10μmol/L was the optional concentration. Under this concentration, the proliferation and ATP level were decreased in Gln(+)+BPTES group (P<0.05), and cellular ROS level was up-regulated compared with Gln(+) group. The survival ratio was significantly lower in BPTES+H 2O2 group compared with BPTES (+) group or H2O2 (+) group. Conclusion Glutamine deficiency inhibits the proliferation and survival ratio of H446 cells through enhancing ROS level. BPTES and H2O2 show synergistically inhibitory effect on the survival of H446 cells.

Objective To construct reference standards for detection and quantification of Klebsiella pneumoniae (K.pneumoniae) with SYBR Green I-based real-time PCR assay. Methods Primers were designed based on the published sequence of the phoE gene of K.pneumoniae. The standard was prepared by cell culture, PCR and T-A clone methods, and was identified by colony PCR and DNA sequencing. Results The standard curve showed a very good linear negative regression between threshold cycle (Ct) and Log starting quantity of copy number. The detection range was from 5.2 to 5.2×106 copies per reaction, and the detection limit was 6 copies per reaction. The coefficients of variance (CVs) of three parallel experiments were in the range of 0.05%-0.91%. Conclusion The reference standards have high stability and reproducibility. They can be used in the quantitative detection of K.pneumoniae.

Objective To evaluate the efficacy and safety of transarterial chemoembolization (TACE)combined with sonographically guided percutaneous microwave coagulation therapy(PMCT)for hepatic carcinoma with diameter＞5.0 cm.Methods We retrospectively reviewed 68 cases of hepatic carcinoma with diameter＞5.0 cm under treatment of TACE combined with PMCT.CT,USG and correlated laboratory tests of hepatic carcinoma were carried out.Results Among 68 cases,complete ablation were 5 cases(5/68),tumor ablation area more than 50% or tumor shrinkage less than 30% were 59 cases(59/ 68),tumor ablation area less than 50% or tumor shrinkage more than 30% were 6 cases(6/68).Forty five cases with high AFP descended more than 50% after the procedure in 42 eases(93.33%).Thirty seven cases and 29 cases with increase of CEA and CA19-9 decreased to 28(75.97%)and 23(93.10%)cases with corresponding index decreasing more than 50% respectively.Survival time reached 4-6 months in 3 cases, more than 6 months for 31 cases,more than 12 months of 34 cases.Two cases among them showed no recurrence up to now after stoppage of treatment for 24 months and finally no correlative mortality occurred. Conclusion TACE combined with sonographically guided PMCT for hepatic carcinoma with diameter more than 5 cm is safe and effective.

AIM: To determine whether the high mobility group protein I (HMGI) is able to bind to the upstream sequence of platelet-derived growth factor B-chain gene and to characterize the HMGI-binding AT-rich regions. METHODS: Recombinant human HMGI (rhHMGI) protein was prepared and electrophoresis mobility shift assay (EMSA) was used. RESULTS: The binding of rhHMGI to PDGF-B (-1 758 / +43 bp) was observed in vitro. Two major HMGI-binding fragments -1 392 / -1 180 bp and -188 / +43 bp were identified, which contained the same AT-rich sequence TTTATAAA (-1 333 / -1 326 bp, -1 314 / -1 307 bp and -30 / -23 bp). An oligonucleotide bound to the TTTATAAA and the GAGACC, the core sequence of the shear stress response element of the PDGF-B, could also bind to the HMGI. Furthermore, HMGI facilitated the binding of NF-?B to the GAGACC in the oligonucleotide. CONCLUSION: The HMGI could bind to the upstream sequence of the PDGF-B gene via the AT-rich sequence TTTATAAA, which may play a role in the transcriptional regulation of the PDGF-B gene.

BACKGROUND: Low back pain and low limb disability are classical symptoms of patients with degenerative lumbar spondylolisthesis who are often treated with operative internal fixation.OBJECTIVE: To observe the improvement of low-back pain and low-limbs disability of patients with degenerative lumbar spondylolisthesis following the treatment with general spine system(GSS).DESIGN: Self control clinical study with patients as subjects.SETTING: Department of Orthopaedics, Peking Union Medical College Hospital.PARTICIPANTS: Sixteen patients with degenerative lumbar spondylolisthesis combined with lumbar stenosis, including 10 males and 6 females with the average age of 58.5 years(ranged from 42 to 72 years) received treatment in our hospital between September 2001 and December 2001. Patients with low back pain were found in 16 cases and low-limb claudication in 15cases; preoperative spondylolisthesis of grade I was observed in 10 cases and grade Ⅱ in 6 cases.METHODS: After receiving lumbar canal decompression by spinal process and GSS for restoring spondylolisthetic vertebra, as well as internalfixation and lumbar fusion operation, patients were followed up at postoperative week 2 and month 1,3,6,12,18,24 for assessing the low-back pain and low-limb disability and meanwhile preoperative spondylolisthesis and postoperative restroration were also assessed with X-ray.RESULTS: Totally 16 patients were followed up for an average of21.2appeared in 15 out of 16 patients, and intermittent claudication resumed to time of follow-up revealed that all spondylolisthetic vertebra were restored with restoring rate of 100% (16/16).CONCLUSION: GSS was proved of satisfactory therapeutic effect in patients with spondylolisthesis by obviously improving their low-back pain and intermittent claudication.

Low frequency sensorineural deafness is a common subtype of idiopathic sudden deafness. Certain patients suffered recurrent attacks without vertigo, much alike Meniere's disease. Few of them developed into definite Meniere's disease during long-term follow-up in many clinical studies. Although the pathophysiology of recurrent low frequency deafness is yet unknown, the desease is considered associated with early state of endolymphatic hydrops or migraine. Otologists shall be aware of its clinical course and careful explanation is necessary at time of initial informed consent.
Endolymphatic Hydrops ; complications ; Hearing Loss, Sensorineural ; complications ; diagnosis ; Hearing Loss, Sudden ; Humans ; Meniere Disease ; complications ; Vertigo

OBJECTIVETo investigate the mechanism of hepatocytes transdifferentiation to bile duct epithelial cells (BECs) and intervention of Huangqi decoction (HQD) on hepatic fibrosis formation in rats with secondary cholestasis.

METHODSSeventy-five SD male rats were made into cholestatic hepatic fibrosis model animals by bile duct ligation, and randomized into the control group (n = 50) and the HQD group (n = 15). Starting from one week after modeling, they were administered orally with saline and HQD respectively for four weeks. Besides, a sham-operated group was set up with 10 rats operated by choledochus segregating only and administered after then with saline. Rats were killed in batches at different time points, i.e. each five from the control group and sham-operated group at the end of the 1st week, five from the control group for each time at the end of the 2nd, 3rd and 4th week, and all the remaining rats at the end of the 5th week. Their liver tissues were taken for histological change examination, content of hydroxyproline (Hyp) determination; protein expression of BECs marker cytokeratin 7 (CK7) and the hepatocyte specific antigen HepPar detection by Western blot, and CK7-Hep Par co-localization by laser confocal microscopy. Then IPP software was used to analyze Sirius red stained positive areas of CK7 and Hep Par, as well as the average IOD of CK7/Hep Par co-localization.

RESULTSHepatocytes in hepatic tissues (Hep Par positive cell) in the model rats decreased gradually along was time went by after modeling (Sham > M1w > M2w > M3w > M4w > M5w), which was in parallel with the increase of BECs (CK7 positive cells), degree of fibrosis, Hyp content and CK7 protein expression. Increasing of co-localized positive cells of CK7/Hep Par began at 1 week and reached the peak 3 weeks after modeling, then it decreased gradually. The Hep Par protein expression was negatively correlated with that of CK7; the Hep Par positive cell expression was negatively correlated with CK7 positive cell expression and collagen deposition; while the CK7 positive cell expression was positively correlated with the collagen deposition in the liver tissue. Compared with the model control group, the mortality, CK7/Hep Par co-localized positive cells, fibrosis degree, Hyp content and CK7 protein expression were lesser obviously (P < 0.01), while Hep Par positive cell and protein expressions were higher significantly in the HQD group.

CONCLUSIONSHepatocytes transdifferentiation to BECs might be a key pathological element for secondary cholestatic hepatic fibrosis formation; the restraining action of HQD is possibly a major action mechanism of HQD for effectively intervening and treating secondary cholestasis hepatic fibrosis.

Animals ; Astragalus Plant ; Bile Ducts ; cytology ; Cell Transdifferentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; Hepatocytes ; cytology ; drug effects ; Liver ; Liver Cirrhosis, Biliary ; drug therapy ; pathology ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley