Four triterpenoids were isolated and purified from the 95% ethanol extract of Maytenus guangxiensis by silica gel column chromatography, Sephadex LH-20 column chromatography, MCI column chromatography and preparative RP-HPLC. Their structures were determined from their physicochemical properties and spectral data. They were identified as maytguanone A (1), maytguanone B (2), 11α-methoxyurs-12-ene-1β,3β-diol (3), lup-20(29)-ene-3β,11α-diol (4). Compounds 1 and 2 are new triterpenoids, along with compounds 3 and 4 were isolated from M. guangxiensis for the first time. The cytotoxicity of compounds 1, 3 and 4 was evaluated using the MTT procedure with three cancer cell lines. The results show that compound 3 displayed good inhibitory effects against HeLa, with an IC₅₀ of 10.68 μmol·L^-1.

OBJECTIVETo develop a colon-specific prodrug of Indomethacin microbially triggered, carry out in vitro/in vivo evaluation of drug release, and appraise its inhibitory effect on liver metastasis from colon cancer.

METHODSIndomethacin prodrugs were synthesized and characterized by FTIR and NMR, and dissolution test simulating gastrointestinal tract was employed to screen the colon-specific prodrug. Then, the pharmacokinetic profile of portal vein and peripheral blood in Sprague-Dawley rats was studied. Lastly, the inhibitory effect on liver metastasis from colon cancer in nude mice was observed.

RESULTSThe chemical structure characterized by FTIR and NMR demonstrated that six kinds of indomethacin-block-amylose with different drug loading (IDM-AM-1-6) were synthesized, among which IDM-AM-3 was degraded 1.3%, 9.3% and 95.3%, respectively, in simulated gastric fluid for 4 h, small intestine for 6 h, and colon for 36 h. The pharmacokinetic test of IDM-AM-3 showed that absorption was delayed significantly (P < 0.01), peak time [(11.35 + or - 2.45) h], elimination half-life [(16.74 + or - 4.04) h] and mean residence time [(22.27 + or - 0.52) h] were significantly prolonged (P < 0.01), as well as peak serum concentrations [(9.69 + or - 2.40) mg/L] and AUC(0-t) [(236.7 + or - 13.1) mg x L(-1) x h] were decreased markedly (P < 0.01) as compared with those of IDM regarding to portal vein. Additionally, its AUC(0-t) in peripheral blood was remarkably lower than that in Portal vein (P < 0.01). The tumor suppression observation showed that it could remarkably reduce the number of liver metastases in contrast to IDM (P < 0.05).

CONCLUSIONColon-specific IDM-AM-3 possesses advantage of sustained release in portal vein providing some experimental basis for colon-specific delivery system applied to sustained release in the portal vein.

Amylose ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Animals ; Colon ; metabolism ; Colonic Neoplasms ; pathology ; Delayed-Action Preparations ; Drug Delivery Systems ; HT29 Cells ; Humans ; Indomethacin ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Prodrugs ; administration & dosage ; chemical synthesis ; pharmacokinetics ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Diagnostic Errors ; Humans ; Knee Joint ; Male ; Middle Aged ; Synovitis, Pigmented Villonodular ; diagnosis ; Tuberculosis, Osteoarticular ; diagnosis

AIMTo determine the effects of azide methyl anthraquinone derivative (AMAD) on growth inhibition and inducing apoptosis of multidrug resistant (MDR) KBv200 cells and parental drug-sensitive KB cells.

METHODSCytotoxicity was determined by tetrazolium (MTF) assay. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (deltapsi(m)) in cells were labeled with DCFH-DA and DiOC6 and tested by flow cytometry. Annexin V stain and DNA ladder were used to examine the apoptosis of KB and KBv200 cells induced by AMAD.

RESULTSAMAD was shown to inhibit the growth of KB and KBv200 cells significantly in a concentration-dependent manner, with mean IC50 of 0.36 and 0.45 micromol x L(-1), respectively. The generation of ROS increased obviously after the cells were treated with AMAD for 12 h, up to the peak in 24 h, meanwhile the levels of deltapsi(m) were time-dependently decreased. DNA fragmentation appeared on the agarose gel. Annexin V stain showed AMAD induced apoptosis of KB and KBv200 cells also in a concentration-dependent manner.

CONCLUSIONAMAD showed inhibitory effect on both MDR KBv200 cells and parental drug-sensitive KB cells. The mechanism of action was associated with the increase of the cellular ROS level and the decrease of the mitochondrial membrane potential induced by AMAD, which result in cell apoptosis.

Anthraquinones ; administration & dosage ; chemistry ; pharmacology ; Antineoplastic Agents ; administration & dosage ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; KB Cells ; Mitochondria ; physiology ; Molecular Structure ; Mouth Floor ; Mouth Neoplasms ; pathology ; Reactive Oxygen Species ; metabolism ; Vincristine ; pharmacology

OBJECTIVETo demonstrate the changes of resting energy expenditure (REE), substrate metabolism and body composition in cancer patients.

METHODSFrom September 2004 to March 2008, REE, carbohydrate oxidation (CO) and fat oxidation (FO) in 936 cancer patients and 840 control subjects were measured by indirect calorimetry. Bioelectrical impedance appliance was applied to assess intracellular fluid, extracellular fluid, fat mass (FM) and fat free mass (FFM) in the two groups.

RESULTSNo difference in REE was found between the cancer patients and non-cancer patients [(1452.2 +/- 196.4) kcal/d vs. (1429.5 +/- 182.6) kcal/d, P = 0.136]. But REE/FFM and REE/pREE were elevated in cancer patients than in controls (all P < 0.05). Of the cancer patients, 48.6% were hypermetabolic, 42.9% normal and 8.5% hypometabolic, while those were 22.5%, 58.5% and 19.0% in controls. Cancer patients had higher FO [(77.8 +/- 11.3) g/min vs. (67.1 +/- 12.1) g/min, P = 0.000], lower CO and npRQ [(68.7 +/- 10.5) g/min vs. (88.8 +/- 12.1) g/min, P = 0.000; 0.782 +/- 0.012 vs. 0.810 +/- 0.014, P = 0.000]. Cancer patients exhibited lower FM and FFM [(14.9 +/- 4.5) kg vs. (18.4 +/- 5.2) kg, P = 0.000; (44.4 +/- 7.2) kg vs. (46.1 +/- 8.1) kg, P = 0.008].

CONCLUSIONSElevated REE is common in cancer patients. Substrate metabolism of the cancer patients features in increased FO, decreased CO and npRQ, which is correlated with the elevated REE. FM and FFM loses in proportion in cancer patients.

Body Composition ; Carbohydrate Metabolism ; Energy Metabolism ; Fats ; metabolism ; Female ; Humans ; Male ; Neoplasms ; metabolism

OBJECTIVETo observe the effect of Chinese drugs for strengthening Pi, harmonizing Wei, and dispersing blood stasis (CDSPHWDBS) on the expression of gastric mucosal heat shock protein 70 (HSP70) in chronic atrophic gastritis (CAG) patients.

METHODSA total of 100 CAG patients were assigned to the control group and the treatment group by random digit table, 50 in each group. Patients in the control group took Folic Acid Tablet 10 mg each time, 3 times per day. Those in the treatment group took CDSPHWDBS, 100 mL each time, once per day. The treatment course was 6 months for all. Clinical symptoms and signs, endoscopic and histopathological changes were observed before and after treatment in the two groups. The expression of gastric mucosal HSP70 in CAG patients was determined using SP immunohistochemistry. Data were collected by HPIAS-1 000 pathological graphic analysis system, and its expression semi-quantitatively analyzed.

RESULTSThe total effective rate of clinical Chinese medical symptoms and signs was 88. 0% (44/50 cases) in the treatment group and 56. 0% (28/50 cases) in the control group, with significant difference between the two groups (P <0. 01). The improvement rate of endoscopic manifestations such as congestion and edema, erosion, bile regurgitation, pale gastric mucosa, exposed blood vessels, particles proliferation in the treatment group were superior to those in the control group (P <0. 05). The total effective rate of atrophy was 80. 0% (40/50 cases) in the treatment group and 54. 0% (27/50 cases) in the control group, with significant difference between the two groups (P<0. 01). The effective rate of intestinal metaplasia was 75. 0% (12/16 cases) in the treatment group and 33.3% (5/15 cases) in the control group, with significant difference between the two groups (P < 0. 05). The optical density value of gastric mucosal HSP70 was significantly elevated in the two groups after treatment (both P <0. 05). It was higher in the treatment group than in the control group after treatment with significant difference (P <0. 01).

CONCLUSIONCDSPHWDBS had obvious effect in treatment of CAG and could improve pathological changes of precancerous lesions possibly by promoting the expression of gastric mucosal HSP70 in CAG patients.

Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis, Atrophic ; drug therapy ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Immunohistochemistry ; Medicine, East Asian Traditional

OBJECTIVETo clone and express Streptococcus suis serotype 2 (S. suis 2) sly gene for constructing an foundation on identification of S. suis 2 protective antigen.

METHODSThe sly gene was amplified from S. suis 2 clinical isolate strain 05ZYH33 genome DNA by PCR. The gene fragment was inserted into the expression vector pET-30b(+) to build pET30b-sly. When recombinant vector pET30b-sly was identified by restriction enzyme cutting and DNA sequencing as a correct one, subsequently it was transformed to E. coli Rosetta for expression under IPTG induction. The obtained fusion protein was purified by Ni-NTA affinity chromatography. The immunologic and hemolysis activity of the purified protein was proved through Western blot and hemolysis assay respectively.

RESULTSThe PCR product was around 1500 bp. The gene segment inserted into the recombinant vector was proven to be completely identical with the sly gene sequence in the total genome sequence of S. suis 2. The target protein expressed was up to 30% of the total somatic protein under IPTG induction. The protein purity reached above 80% after purification. The protein could be recognized by human serum infected with S. suis 2 and could dissolve swine erythrocytes with the Hemolytic titer as 256.

CONCLUSIONThe expression vector pET30b-sly was successfully constructed. The target protein could be over-expressed in E. coli and possessed its biological activity after purification.

Animals ; Bacterial Proteins ; genetics ; metabolism ; pharmacology ; Blotting, Western ; Chromatography, Affinity ; Hemolysis ; drug effects ; Humans ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; Streptococcus suis ; genetics ; metabolism ; Swine

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

In this study,we aimed to study the pattern visual evoked potentials (P-VEPs) in two eyes with varying visual acuity in one eye and to provide an objective estimation of visual acuity by comparing P-VEPs in one and two eyes.Thirty subjects were chosen,who had one eye with an acuity of 5.0,4.85,4.6,4.0,or scieropia and obstructed vision and the other eye with an acuity of 5.0,respectively.P-VEPs were detected under the large grating stimuli at 3×4 spatial frequency,moderate grating stimuli (12× 16 spatial frequency) and small grating stimuli (48×64 spatial frequency).Under large grating stimuli,there was no significant difference in P100 peak latency between the groups,nor was there a significant difference between the amplitude of two eyes and the amplitude of one normal-vision eye.Under moderate and small grating stimuli,there was a significant difference in P100 peak latency between the group with both eyes having an acuity of 5.0 and the group with visual acuity below 4.0 in one eye.There was a significant difference in P100 amplitude between the group with visual acuity of 5.0 in both eyes and the group with one normal-vision eye.There was no significant difference in the amplitude of two eyes and the amplitude of one normal-vision eye between any other two groups.In forensic identification,characteristics and variability of P-VEPs in one and two eyes can be used to identify malingering or decline in visual acuity.

Objective To explore the genetic interactions between angiotensin-convertingenzyme (ACE) I/D and methylenetetrahydrofolate reductase (MTHFR) C677T genotypes in middlecerebral artery stenosis (MCAS) among the asymptomatic residents in Foshan area of China. MethodsUsing a cluster sampling method, 2500 subjects were randomly selected from the residential communitiesof Rongqi town of Foshan area, Guangdong Province. By means of epidemiological questionnaire survey,physical examination, examination of the biochemical markers and transcraniai color Doppler (TCD), 897eligible subjects (306 males and 591 females) were selected from this population and subsequentlydivided into MCAS group and control group according to the TCD results. ACE and METHFR genepolymorphism analyses were conducted using amplified fragment length polymorphism-polymerase chainreaction (AFLP-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Chi-square test, t test, Mann-Whitney test and logistic regression analysis were used to analyze the data. ResultsGender, age, waist-to-hip ratio (WHR) and Ⅱ+CC genotype distribution in the subjects with MCAS weresignificantly different from those in the control subjects. Logistic regression analysis identified age andACE Ⅱ+ MTHFR CC genotype as the independent factors that affected MCAS. Conclusion There aregenetic interactions between ACE I/D and MTHFR C677T genotypes, and the ACE Ⅱ+MTHFR CCgenotype is an independent genetic factor for protection against MCAS in the asymptomatic residents inFoshan area of China.