AIMTo observe expressions and changes of Tau protein, pSer202 and Tau protein's contents during the differentiation process of bone-marrow mesenchymal stem cells (MSCs) into neural cells, and discuss Tau's effects on it.

METHODSEGF and bFGF were combined for the induction of 4th, 8th, and 12th-MSCs into neural cells. Expressions of Tau protein and pSer202 were tested by immunocytochemistry. ELISA assay was applied for testing Tau protein's contents during differentiation process.

RESULTSPositive rates of Tau protein in uninduced MSCs of 4th, 8th, and 12th-MSCs were under < 6%; After 14-day induction, the cellular morphologic characteristics in different passages were very similar to neurons, positive rates of Tau protein had no significant differences between passages (P > 0.05), but had differences with their uninduced groups (P < 0.05). There hadn't had expression of pSer202 in uninduced and induced groups of passages. ELISA assay indicated that there was an upward tendency in Tau protein's contents during the 14-day induction process, those in the 14th day had no significant differences between passages too (P > 0.05).

CONCLUSIONThe increase in Tau protein's expressions and its non-phosphorylated state may make for MSCs differentiating into normal neural cells and formation of neuronal processes.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Guinea Pigs ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; tau Proteins ; metabolism

OBJECTIVETo study the chemical constituents of Ervatamia hainanensis.

METHODThe compounds were separated and purified by column chromatography with silica gel, and identified by IR, MS, NMR and 2D-NMR.

RESULTFive compounds were identified as I (isolariciresinol 9-O-beta-D-glucopyranoside), II (cycloartenol), III (beta-amyrin acetate), IV (beta-sitosterol), V (daucosterol), respectively.

CONCLUSIONAll the compounds were isolated from this plant for the first time.

Apocynaceae ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Phytosterols ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes

OBJECTIVETo investigate the feasibility and safety of the treatment for thoracolumbar fractures with pedicle screw at the fracture level and vertebroplasty via paraspinal approach.

METHODSFrom August 2007 to August 2010, 22 old patients with thoracolumbar fractures were treated with pedicle screw at the fracture level and vertebroplasty via paraspinal approach. There were 14 males and 8 females, ranging in age from 60 to 71 years (mean, 64.6 years). The time from injury to surgery varied from 1 to 4 d (mean,2.7 d). All the patients suffered from single thoracolumbar fractures and located at T11 in 2 cases, at T12 in 5 cases, at L1 in 11 cases and at L2 in 4 cases. According to the Denis fracture classification, there were 6 compression fractures and 16 burst fractures. The mean preoperative load-sharing classification of spine fractures was 5.4 score. The mean preoperative thoracolumbar injury classification and scoring was 5.2. Based on the ASIA neurologic grading system, preoperative neurological function was grade B in 2 cases,grade C in 3 cases, grade D in 7 cases and grade E in 10 cases. The neurological function, vertebral central and anterior height, kyphotic angle of the vertebral fractures by radiographs and visual analog scale were calculated pre-operatively, post-operatively and at the last follow-up.

RESULTSMedian operating time was 60.8 min (ranged from 50 to 95 min) and median blood loss was 84 ml (ranged from 50 to 130 ml). The operative incisions were healed well. The duration of follow-up averaged 21.6 months (ranged from 12 to 48 months). The anterior vertebral body height was corrected from preoperative (52.3 +/- 10.3) % to postoperative (6.1 +/- 4.2) % and (6.8 +/- 5.4) % at the last follow-up. The central vertebral body height was corrected from preoperative (38.9 +/- 11.2) % to postoperative (8.3 +/- 4.7) % and (9.4 +/- 4.5)% at the last follow-up. The Cobbs angle of the injured vertebral segment was corrected from preoperative (19.5 +/- 9.5) degrees to postoperative (4.3 +/- 4.1) degrees and (6.2 +/- 4.7) degrees at the last follow-up. The VAS scores reduced from preoperative 8.56 +/- 0.88 to post-operative 3.48 +/- 0.91 and 3.20 +/- 0.92 at the last follow-up. The postoperative neurologic function of all 22 patients improved 1 to 2 degrees except 10 patients of grade E. There were no instances of instrumentation failure and no patient had persistent postoperative back pain.

CONCLUSIONThe pedicle screw at the fracture level and vertebroplasty via paraspinal approach has the advantages of less invasive and blood loss, and could prevent the development of kyphosis and offers improvement of the spinal cord function. Furthermore, it could decrease the risks of postoperative back pain and the failure of instrumentation.

Aged ; Bone Screws ; Female ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; injuries ; surgery ; Male ; Middle Aged ; Recovery of Function ; Retrospective Studies ; Spinal Cord ; physiopathology ; Spinal Fractures ; diagnostic imaging ; physiopathology ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Vertebroplasty ; instrumentation

Objective To explore the effect of SA liposome mediated human interleukin-10 (IL-10) gent transfection on NHE-1 mRNA expression in penumbra area following focal cerebral ischemia reperfusion injury in rats. Methods Totally 78 male SD rats were randomly divided into normal control group (n=6), MCAO group (n=24), hIL-10 transfection group (n=24) and empty vector transfection group (n=24). Longa's method was employed to establish MCAO models in the latter 3 groups. The rats in the MCAO group underwent stereotactic operation without drug injection, and the hIL-10 transfection group and empty vector transfection group were injected stereotactieally with pcDNA3.1-IL-10 and pcDNA3.1, respectively, both by SA liposome mediation. After transfection, RT-PCR and ELISA were used to determine the effect of transfection, TTC staining was conducted to detect the infarct volume. Meanwhile, real-time quantitative PCR was performed to examine the expressions of NHE-1 mRNA and NF-κB mRNA in the penumbra area. Results (1) SA liposome effectively mediated the hIL-10 gene to transfect the brain tissue. Also hIL-10 gene transfection played neuroprotective effect by reducing the brain infarct volume. (2) The expression of NF-κB mRNA in different groups was 1.00±0.33, 4.76±0.41, 4.58±0.62 and 2.77±0.43, respectively, hIL-10 gene transfection also inhibited the increase of NF-κB mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. (3) The expression of NHE-1mRNA was 1.00±0.22, 4.16±0.48, 3.97±0.51 and 2.82±0.47, respectively, hIL-10 gene transfection also inhibited the increase of NHE-1 mRNA expression in the penumbra area following the cerebral ischemia reperfusion injury. Conclusions The hIL-10 transfection can exert the protective effect on the brain against cerebral ischemia-reperfusion injury partly via inhibiting the NHE-1 mRNA expression.

This study was purposed to investigate the effect of chemotherapeutic drug cyclophosphamide (CTX) on normal murine bone marrow hematopoietic cells, especially on the self-renewal, proliferation and differentiation of bone marrow hematopoietic cells, and possible mechanisms. The CTX-treated mouse model was established by CTX 200 mg/kg, ip. The exact time of complete recovery of hematopoiesis was determined by monitoring the recovery level of differential blood counts and the proportion of LKS(+) cells in bone marrow cells. The function of bone marrow hematopoietic cells such as self-renewal, proliferation and differentiation were assessed by non-competitive and competitive bone marrow transplantation. The potential effect of CTX on senescence of bone marrow hematopoietic cells was analyzed by detecting p16(Ink4a) mRNA relative expression and SA-β-galactosidase (gal) staining. The results showed that the CTX could induce long-term but latent damage to bone marrow hematopoietic cell function and lead to the decrease in competency of bone marrow hematopoietic cells to reconstitute while seemingly permitting a complete recovery. Furthermore, the serial-transplantation model showed that these mice received transplantation of bone marrow hematopoietic cells from CTX-treated mice exhibited a high expression of p16(Ink4a) mRNA and SA-β-gal staining. It is concluded that CTX-induced bone marrow cellular senescence may play an important role in CTX-induced long-term injury to bone marrow hematopoietic cells.
Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cellular Senescence ; drug effects ; Cyclophosphamide ; adverse effects ; Hematopoiesis ; drug effects ; Mice ; Mice, Inbred C57BL

The aim of this study was to explore cytogenetic characteristics of acute promyelocytic leukemia (APL) and compare the interphase fluorescence in situ hybridization (I-FISH) with conventional cytogenetic (CC) analysis. A total number of 157 APL patients were recruited in this study, and the I-FISH and CC were applied to analyze cytogenetic features. Chromosome samples of bone marrow cells were prepared by short-term culture. Out of all 157 cases, 136 were observed with CC assay, 66 with I-FISH, of which 45 samples were analyzed with both methods. The results showed that among all 136 CC samples, t(15;17)(q22;q21) was found in 120 cases, of which 107 cases was isolated t(15;17)(q22;q21) abnormality, 13 cases was complex abnormalities and 12 case without mitotic figure. Among all 66 cases of I-FISH group, PMI/RARα fusion gene was found in 64 cases (97.0%), suggesting that I-FISH group was more sensitive than CC group (p = 0.041). It is concluded that combination of I-FISH and CC techniques plays a pivotal role for diagnosis and detection of minimal residual disease in APL.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Cytogenetic Analysis ; methods ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Promyelocytic, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Young Adult

To test the European BIOMED-1 Concerted Action proposed technique to detect minimal residual disease (MRD) in the chinese patients with precursor-B-acute lymphoblastic leukemia (precursor-B-ALL) by triple-staining flow cytometry and to define both normal and aberrant phenotypic profiles of precursor B cells, a series of bone marrow samples, 35 from precursor-B-ALL (13 in newly diagnosed cases, 15 at the end of remission induction therapy and 7 at end of the consolidations), and 19 from normal controls, were immunophenotyped with the five triple-staining antibodies (TdT/CD10/CD19, CD10/CD20/CD19, CD34/CD38/CD19, CD34/CD22/CD19 and CD19/CD34/CD45) recom-mended by the BIOMED-1 using common flow cytometric protocols. Further, with different ratios of the leukemic cells with CD34/CD38/CD19 phenotype and normal mononuclear cells, a serial dilution test was analyzed. The results showed that three major CD19(+) cell subpopulations were identified in the normal controls, representing three consecutive maturation stages. The subpopulations in the precursor-B-ALL cases disappeared and were replaced with a great number of luekemic cells which had different characteristics of phenotypes, and then they reappeared with almost same characteristics as the normal CD19(+) cells after the patients achieved complete remission. When the five triple-staining antibody combinations were used, the phenotypic aberrancies could be identified in 12/13 (92.3%) cases with newly diagnosed precursor-B-ALL, at least one triple-labeling per case at the level of 0.01% or more. The frequencies of phenotypic aberrations detected with the triple-staining were 8/13 (61.5%) for CD10/CD20/CD19, 5/13 (38.5%) for CD34/CD38/CD19, 4/13 (30.8%) for CD10/TdT/CD19, 3/13 (23.1%) for CD34/CD22/CD19, and 2/13 (15.4%) for CD34/CD45/CD19. At the end of remission induction, the phenotypic aberrancies could be detected in 5/15 (33.3%), of which, 3/8 (37.5%) cases with the leukemic phenotypes detected both at the newly diagnosis and at the end of induction. The dilution test indicated that the cells with CD34/CD38/CD19 detected by flow cytometry correlated well with the leukemic cells added (r = 0.85, P < 0.05) over 1:1 to 1:400,000. It is concluded that the flow cytometric detection of precursor-B-ALL-MRD proposed by BIOMED-1 Concerted Action were well realized in this study. The one precursor-B-ALL cell can be effectively detected out of 10(4) normal bone marrow cells.
Adolescent ; Adult ; Aged ; Antigens, CD ; analysis ; B-Lymphocytes ; immunology ; pathology ; Bone Marrow Cells ; immunology ; pathology ; Child ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Neoplasm, Residual ; blood ; immunology ; pathology ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; blood ; immunology ; pathology

OBJECTIVETo investigate the prevalence and risk factors of diabetic retinopathy (DR) among type 2 diabetic patients aged over 30 in Shanghai central area.

METHODS1039 patients diagnosed with type 2 diabetes mellitus (DM) aged over 30 were investigated by randomized cluster sampling in Shanghai central area and data from 767 of those patients were analyzed.

RESULTS(1) Among all of the 1534 digital ocular fundus images from 767 patients, 87.6% of the images from 672 patients were gradable. (2) Among all of the 672 patients with gradable ocular fundus images, the prevalence of non-proliferative diabetic retinopathy (NPDR) was 21.6%, while proliferative diabetic retinopathy (PDR) was 1.3%. The rates of mild, moderate and severe NPDR were 8.8%, 11.2% and 1.6% respectively. (3) DR patients were characterized with elder age, higher HbA1c, urea nitrogen and serum creatinine. DM duration and the level of fasting plasma glucose were risk factors for DR.

CONCLUSIONThe overall prevalence of DR in type 2 diabetic patients aged over 30 in Shanghai central area was 22.9% and the DR risk factors were found to include duration of diabetes and fasting plasma glucose level.

Aged ; China ; epidemiology ; Cluster Analysis ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Retinopathy ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Risk Factors

BACKGROUND/AIMS: Methylation status plays a causal role in carcinogenesis in targeted tissues. However, the relationship between the DNA methylation status of multiple genes in blood leukocytes and colorectal cancer (CRC) susceptibility as well as interactions between dietary factors and CRC risks are unclear. METHODS: We performed a case-control study with 466 CRC patients and 507 cancer-free controls to investigate the association among the methylation status of individual genes, multiple CpG site methylation (MCSM), multiple CpG site heterogeneous methylation and CRC susceptibility. Peripheral blood DNA methylation levels were detected by performing methylation-sensitive high-resolution melting. RESULTS: Total heterogeneous methylation of CA10 and WT1 conferred a significantly higher risk of CRC (adjusted odds ratio [OR(adjusted)], 5.445; 95% confidence interval [CI], 3.075 to 9.643; OR(adjusted), 1.831; 95% CI, 1.100 to 3.047; respectively). Subjects with high-level MCSM (MCSM-H) status demonstrated a higher risk of CRC (OR(adjusted), 4.318; 95% CI, 1.529 to 12.197). Additionally, interactions between the high-level intake of fruit and CRH, WT1, and MCSM on CRC were statistically significant. CONCLUSIONS: The gene methylation status of blood leukocytes may be associated with CRC risk. MCSM-H of blood leukocytes was associated with CRC, especially in younger people. Some dietary factors may affect hypermethylation status and influence susceptibility to CRC.
Carcinogenesis ; Case-Control Studies ; China* ; Colorectal Neoplasms* ; DNA Methylation ; Freezing ; Fruit ; Humans ; Leukocytes ; Methylation* ; Odds Ratio

Objective: To investigate the effect of varying concentrations of polyethylene glycol(PEG)400 in receiving solution on in vitro transdermal test of drugs. Method: 5-Fluorouracil(5-FU) was selected as a model drug,by preparing different concentrations of PEG400-phosphate buffer solution(PBS) as the receiving solution,the receiving chamber did not add drug,the excised rat skins were treated with various additives for 12 h,then replaced by PBS and added the saturated model drug into the donor compartment to determine the transdermal parameters of the drug.Meanwhile,scanning electron microscopy(SEM) was employed to monitor the effect of PEG400 with different concentration on the stratum corneum of rat skin. Result: The 10%,15% and 40% PEG400-PBS groups had no significant effect on in vitro transdermal absorption parameters of the 5-FU.The steady transdermal rate and cumulative penetration rate of the drug in 20% and 30% PEG400-PBS groups were significantly higher than that in the PBS group(P<0.01,P<0.05).SEM indicated that wrinkle of the intact rat skin gradually disappeared and a number of flakes were desquamated from the skin when the concentration of PEG400 was above 20% in receiving solution.Meanwhile,30% PEG400-PBS group and 40% PEG400-PBS group were extremely wrinkled. Conclusion: In the rat skin transdermal test,the concentration of PEG400 in receiving solution should be controlled below 20%.