OBJECTIVETo approach the mechanisms of effects of high glucose on the differentiation of periodontal ligament cells(PDLC) by investigating the changes of Scleraxis mRNA expression in high glucose condition in vitro.

METHODSHuman PDLC were cultured in high glucose medium (4500 mg/L glucose) and normal glucose medium (1000 mg/L glucose), respectively. High glucose was used to inhibit the osteogenic differentiation of PDLC. PDLC cultured in normal glucose medium served as control. Alkaline phosphatase (ALP) activity, the early parameter of osteogenetic differentiation of cells and the expression of Scleraxis mRNA were detected in each group. ALP activity was measured colorimetrically by using nitrophenyl phosphate as a substrate and the expression of Scleraxis mRNA was analyzed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).

RESULTSALP activity of PDLC was lower in high glucose medium than in normal glucose medium, and the values were 0.113 +/- 0.068 and 0.218 +/- 0.012, respectively. However, the level of Scleraxis mRNA was quite higher in high glucose medium compared with in normal glucose medium, and the values were 0.973 +/- 0.055 and 0.611 +/- 0.205, respectively. The values of ALP activity and the expression of Scleraxis mRNA were significantly different between the two groups.

CONCLUSIONSHigh glucose inhibited osteogenetic differentiation of PDLC and up-regulated Scleraxis expression. The adverse changes of Scleraxis expression and osteogenic differentiation of PDLC suggest that Scleraxis may regulate the osteogenic differentiation of PDLC negatively and the inhibition of high glucose on osteogenetic differentiation of PDLC may be regulated by Scleraxis in transcription level.

Adolescent ; Adult ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Differentiation ; Cells, Cultured ; Culture Media ; Glucose ; metabolism ; pharmacology ; Humans ; Periodontal Ligament ; cytology ; drug effects ; metabolism ; Young Adult

Objective: To observe the clinical effects of moxibustion at abdominal acupoints for slow transit constipation (STC) due to yang deficiency of the spleen and kidney. Methods: A total of 52 cases with slow transit constipation in conformity with the inclusion criteria were selected and divided into a control group and an observation group according to their visit order and random digital table, 26 cases in each group. Patients in the control group received routine nursing guide. Besides the same routine nursing guide, patients in the observation group received moxibustion at the abdominal acupoints, once every day. The course of the treatment was 4 weeks in the two groups, and the 3-month follow-up was given after the course was finished, for comparing the clinical symptoms, results of colon transit tests, scores of depression/anxiety scale and nursing satisfaction. Results: The total effective rate was 92.3% in the observation group and 69.2% in the control group, with a significant difference between the two groups. After the treatment and during the follow-up checks, the scores of Chinese medical symptoms in the two groups were remarkably decreased than those before the treatment (all P<0.01); the scores of the observation group were obviously lower than those in the control group (all P<0.01). The discharge rates of the markers in the two groups were remarkably increased than those at the same time period before the treatment; moreover, the discharge rates of the markers at various time periods were remarkably better in the observation group than those in the control group (P<0.01). SDS and SAS scores were remarkably decreased after the treatment in the two groups (P<0.01). In comparison of SDS score between the two groups after the treatment, the difference was statistically significant (P<0.01). In comparison of SAS score between the two groups after the treatment, the difference was statistically significant (P<0.05). The nursing satisfaction was 96.2% in the observation group after the treatment, obviously better than that in the control group (73.1%). The recurrence rate was 8.3% in the observation group, remarkably lower than that in the control group (33.3%). Conclusion: Moxibustion at the abdominal acupoints plus routine nursing can remarkably improve the colon transit functions and anxious and depressive emotion in patients with STC, and the therapeutic effects are remarkable. Not only the clinical satisfaction is higher, but the recurrence rate is obviously lower than that of routine nursing.

Three compounds were isolated from the extract of Taxus cuspidta Sibe et Zucc with the column chromatography on silica gel and preparative HPLC methods. Their structures were identified according to the physicochemical properties and spectral analysis, and they were identified as (E)-1-methoxy-2-O-(p-coumaroyl)-myo-inositol (1), 2-deacetoxy-7beta, 9a, 10beta-trideacetyltaxinine J (2) and (3aS, 4aR, 6S, 8S, 8aS, 9R, 10R, 10aS)-benz[f]azulene-6, 8, 9, 10 (3H)-terol, 3a, 4, 4a, 5, 6, 7, 8, 8a, 9, 10-decahydro-10a-(1-hydroxyl-1-methylethyl)-1, 8a-dimethyl-5-methylene (3). Among them, compound 1 was a new compound, and compounds 2, 3 were two novel natural products.
Azulenes ; chemistry ; isolation & purification ; Coumarins ; chemistry ; isolation & purification ; Inositol ; analogs & derivatives ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Taxoids ; chemistry ; isolation & purification ; Taxus ; chemistry

To investigate the difference in absorptive of tetrahydropalmatine (THP) and l-tetrahydropalmatine (l-THP) in rat intestine as well as the mechanism of the absorption of THP, in situ single pass perfusion model was used and the concentration of THP in perfusate was determined by HPLC. The absorption rate constant (k(a)) and effective permeability values (P(eff)) of THP had no significant difference (P > 0.05) at concentration of 8, 16 and 32 microg x mL(-1) in perfusion or in four different regions of intestine of rat (duodenum, jejunum, ileum, colon). The absorption of l-THP and THP in jejunum had significant difference (P < 0.05). The k(a) and P(eff) of THP increased obviously when verapamil was co-perfused with THP, while those of l-THP were not influenced by verapamil. The absorption of THP in intestine showed the passive diffusion process, and without a special absorption region. The stereoselective absorption difference may result from stereoselective combination of P-glycoprotein with d-THP.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Berberine Alkaloids ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Duodenum ; metabolism ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Jejunum ; metabolism ; Male ; Perfusion ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sex Factors ; Stereoisomerism ; Verapamil ; pharmacology

OBJECTIVETo study on needling safe depth of Fengfu (GV 16) with CT, so as to provide reference for safe needling depth of Fengfu (GV 16) in clinical acupuncture treatment.

METHODSForty-one adult volunteers were divided into 3 groups, a thin person group, a moderate person group and a fat person group according to Luo's indexes, and computer-aided tomography (CT) was used to measure the needling depth of Fengfu (GV 16).

RESULTSThe safe depths of perpendicular needling of Fengfu (GV 16) were different for persons of different somatotypes. The safe needling depth was (27.73 +/- 3.45) mm for the thin person group, (30.78 +/- 2.90) mm for the moderate person group, and (33.39 +/- 4.27) mm for the fat person group.

CONCLUSIONThe safe needling depth < or = the dangerous depth x 75% can be used for reference for the safe needling depth of Fengfu (GV 16) for different somatotypes persons.

Acupuncture Points ; Acupuncture Therapy ; Female ; Humans ; Male ; Neck ; Somatotypes ; Tomography, X-Ray Computed

Objective To observe the clinical efficacy of moxibustion at He-Sea plus Front-Mu points in treating chronic atrophic gastritis (CAG) due to deficient cold in spleen-stomach, and its effect on serum pepsinogen (PG) and gastrin. Method Sixty-three eligible patients with CAG due to deficient cold in spleen-stomach were divided into a control group (31 cases) and a treatment group (32 cases) by using random number table. The control group was intervened by Wei Fu Chun tablets, while the treatment group received moxibustion at He-Sea plus Front-Mu points, 12 weeks as a treatment course. Clinical efficacy, symptoms score of traditional Chinese medicine (TCM), serum PGⅠ, PG Ⅱ and gastrin levels were evaluated. Result The total effective rate was 93.8% in the treatment group, significantly better than that in the control group (P<0.05); the TCM symptoms scores dropped significantly in both groups after the treatment and in the follow-up study (P<0.01), and the treatment group was markedly superior to the control group (P<0.01); the levels of PG Ⅰ, PG Ⅰ/Ⅱ ratio (PGR) and gastrin-17 (G-17) increased significantly in the treatment group after the intervention (P<0.01), and PG Ⅱ dropped significantly (P<0.05); after the treatment, thelevels of PG I, PGR and G-17 increased significantly in the control group (P<0.05); after the intervention, the treatment group was significantly better than the control group in comparing the levels of PG Ⅰ, PGR and G-17 (P<0.01), and there was a significant difference in comparing the level of PG Ⅱ between the two groups after the intervention (P<0.05). The follow-up showed that the HP positive rate was 3.1% in the treatment group, significantly lower than that in the control group (P<0.05). Conclusion Moxibustion at He-Sea plus Front-Mu points can significantly improve CAG symptoms, enhance HT clearance and lower the relapse; its action mechanism is possibly through up-regulating PGⅠ, PGR and G-17 and down-regulating PG Ⅱ.

Objective: To observe the effects of herb-partitioned moxibustion and ginger-partitioned moxibustion on the growth of colon tumors in rats with colitis-associated colon cancer (CACC), and explore the mechanism of moxibustion intervening CACC through the purinergic receptor P2X ligand-gated ion channel 7 (P2X7R)/signal transducer and activator of transcription 3 (STAT3)/vascular endothelial growth factor (VEGF) pathway. Methods: A total of 26 male Sprague-Dawley rats were selected. According to the random number table method, 6 rats were selected as the normal group. The remaining 20 rats were injected intraperitoneally with azoxymethane (AOM) combined with oral dextran sodium sulfate (DSS) to prepare the CACC model. After the model was successfully established, 2 rats were randomly selected for model identification. The remaining 18 rats which were successfully modeled were randomly divided into a model group, a herb-partitioned moxibustion group and a ginger-partitioned moxibustion group, with 6 rats in each group. Moxibustion intervention was performed in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group at Qihai (CV 6) and bilateral Tianshu (ST 25). Moxibustion was performed twice at each point each time, once a day, at a 1-day interval after 6 consecutive interventions, for a total of 30 interventions. After intervention, the colon tumor load, pathological change and histopathological score were observed. Immunohistochemistry was used to detect the expressions of VEGF, P2X7R, phospho-STAT3 (p-STAT3), and nuclear factor-kappa B p65 (NF-κB p65) proteins in rat colon tissue. Western blot was used to detect the levels of p-STAT3 and NF-κB p65 proteins in rat colon tissue. Results: Compared with the normal group, the colon tumor load and histopathological score in the model group were significantly increased (both P<0.001), and different grades of dysplasia were observed in colon tissue from the model group, reaching the degree of adenocarcinoma; the expression level of P2X7R protein in colon tissue was significantly decreased (P<0.001), and the expression levels of p-STAT3, NF-κB p65 and VEGF proteins were significantly increased (all P<0.001) in the model group. Compared with the model group, the colon tumor load, colon histopathological score and the levels of p-STAT3, NF-κB p65 and VEGF proteins in colon tissue were significantly decreased (all P<0.05) in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group while the expression levels of P2X7R protein in colon tissue were significantly increased (both P<0.05). Conclusion: Both herb-partitioned moxibustion and ginger-partitioned moxibustion can reduce the colon tumor load in CACC rats and delay the progression of colon adenomas. The mechanism may be mediated by the P2X7R/STAT3 pathway to inhibit STAT3 phosphorylation, thereby reducing VEGF protein expression.

OBJECTIVETo investigate the reactivity of colon cancer cell line SW480 and CD133(+) SW480 subsets to hypoxia in vitro and the changes in the expressions of anti-apoptosis and angiogenesis genes.

METHODSSW480 cells was subjected to CoCl(2) exposure at varying concentrations and for different time lengths to induce hypoxia, and the protein expression of hypoxia induced factor 1α (HIF-1α) was detected by Western blotting. The CD133(+) SW480 cells were sorted by magnetic activated cell sorting (MACS) and their proportion was assayed by flow cytometry (FCM). The CD133(+) SW480 subsets were exposed to CoCl(2) at the optimal concentration with exposure time selected in terms of HIF-1α level, and their tumor stem cell sphere formation ability was evaluated. Real-time PCR was used to compare the mRNA expression levels of the surface markers of colon cancer stem cells (CD133 and PROM1), survivin, and vascular endothelial growth factor (VEGF).

RESULTSExposure to 200 µmol/L CoCl(2) for 8 h resulted in the highest HIF-1α expression in SW480 cells, but the same exposure failed to induce HIF-1α expression in CD133(+) SW480 subsets. The CD133(+) SW480 subsets, after CoCl(2)-induced hypoxia, showed significantly enhanced ability of cell sphere formation. Hypoxia of SW480 cells caused significant increases in CD133, survivin and VEGF mRNA levels by 1.607∓0.103, 2.745∓0.370 and 3.798∓0.091 folds, respectively (P<0.05).

CONCLUSIONCoCl(2) can simulate hypoxia in colon cancer cells in vitro to induce stable HIF-1α expression, which is concentration- and time-dependent. The hypoxia-stimulated tumor stem sells show an enhanced sphere formation and anti-apoptotic and anti-angiogenic abilities.

Apoptosis ; physiology ; Cell Hypoxia ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; Computer Simulation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Neoplastic Stem Cells ; pathology ; Neovascularization, Pathologic ; physiopathology

OBJECTIVETo study the clinicopathologic features, immunophenotype and prognosis of primary cutaneous anaplastic large cell lymphoma (C-ALCL).

METHODSEight cases of C-ALCL were enrolled into the study. The clinicopathologic features, immunohistochemical findings and results of in-situ hybridization for EBER 1/2 were analyzed.

RESULTSThree of the 8 patients were males and 5 were females. The median age was 49.5 years. C-ALCL often presented with solitary skin nodule, without systemic symptoms. Histologically, the lymphoma cells infiltrated the dermis and subcutis in a sheet-like pattern. They were of large size and showed conspicuous nuclear atypia. Immunohistochemical study showed that more than 75% of the lymphoma cells were positive for CD30. All cases expressed one to three T cell markers (CD3, CD5 or CD45RO) and cytotoxic granule-associated antigens (TIA-1, granzyme B or perforin). The staining for leukocyte common antigen was positive in all cases, while the expression of CD5, CD8, ALK-1 and epithelial membrane antigen was noted in 5, 1, 1 and 3 cases, respectively. The staining for CD15, CD20, CK and HMB45 was negative. In-situ hybridization for EBER 1/2 was also negative in all the cases studied. Follow-up information was available in 6 patients. Five of them were still alive and 1 died of unclear cause.

CONCLUSIONSC-ALCL has distinctive clinicopathologic and immunophenotypic features. It is not Epstein-Barr virus-related and often carries a favorable prognosis.

Adult ; Aged ; CD5 Antigens ; metabolism ; Child ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Immunophenotyping ; In Situ Hybridization ; Ki-1 Antigen ; metabolism ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Primary Cutaneous Anaplastic Large Cell ; immunology ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Prognosis ; RNA, Viral ; metabolism ; Skin Neoplasms ; immunology ; metabolism ; pathology ; therapy ; Young Adult

OBJECTIVETo investigate the clinicopathologic features of lymphoplasmacytic lymphoma (LPL) with Waldenström's macroglobulinemia (WM) and to evaluate the usefulness of immunophenotype analysis in diagnosis and differential diagnosis of the tumor.

METHODSA total of 40 cases of LPL with WM diagnosed according to the 2008 World Health Organization classification of tumors of hematopoietic and lymphoid tissues were analyzed using immunophenotype and follow-up information.

RESULTSThe mostly common initial clinical presentations were non-specific symptoms, such as fatigue, anemia and hemorrhage. Lymphadenopathy, splenomegaly and hepatomegaly were found in 42.5%, 20.0% and 12.5% of the patients respectively. The pattern of bone marrow involvement included mixed type (47.2%), diffuse type (41.7%) and interstitial type (11.1%). The nodal architecture was completely destroyed in one case and partially effaced with residual germinal centers and dilated sinuses in 8 cases. All of the neoplastic cells expressed CD20 and CD79a. Neoplastic plasma cells were positive for CD138 and CD79a. No cases expressed CD5. Four cases weakly expressed CD23. No significant prognosis related factors were identified in the survival analysis.

CONCLUSIONSLPL with WM is a rare indolent small B-cell lymphoma, which is commonly seen, in older male patients. The tumor frequently involves bone marrow and shows various clinical manifestations. Combination analyses of the bone marrow biopsy histology, immunophenotypic study and clinical data, especially the serum examination are important for the diagnosis of LPL with WM.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD20 ; metabolism ; Bone Marrow ; metabolism ; pathology ; CD79 Antigens ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Immunoglobulin M ; blood ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphatic Metastasis ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; pathology ; Lymphoma, Follicular ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; metabolism ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology ; Neoplasm Invasiveness ; Survival Rate ; Syndecan-1 ; metabolism ; Waldenstrom Macroglobulinemia ; immunology ; metabolism ; pathology