AIM: To investigate the apoptosis of a vascular smooth muscle cell line A10 caused by mild K+ depolarization. METHODS: Apoptosis was evaluated by nuclear staining, DNA fragmentation gel electrophoresis and propidium iodide-stained flow cytometry. Mitochondrial transmembrane potential (Δψm) was measured by flow cytometry. RESULTS: K+ depolarization caused dose correlated A10 cells apoptosis; nifedipine, BAPTA/AM, ryanodine inhibited the cytotoxic effect of K+ completely.The combination use of nifedipine and cyclosporin A made it clear that mitochondria was involved in the apoptosis of A10 cells,and Δψm measurement further confirmed this speculation; A10 apoptosis caused by K+ depolarization was not influenced by heparin or Zn2+,a effective capacitative calcium entry(CCE) blocker. CONCLUSION: Ca2+ entry through voltage-dependent ca channels increases intracytoplasm Ca2+, then triggers further Ca2+ release from endoplasmic reticulum via ryanodine receptor, and the microdomains of elevated intracytoplasm Ca2+ are sensed by adjacent mitochondria, which ultimately lead to cell apoptosis.

A low calcium medium developed for epidermal keratinocytes were prepared according to the MCDB 153 modified formula and used in human epidermal keratinocyte culture compared with DMEM culture system. The observation by contrast microscopy and electron microscopy showed that in the low calcium medium keratinocytes grew as a monolayer of high proliferation and had many characteristics of basal cells, with a more rounded shape and large intercellular spaces. Increasing the calcium ion concentration in the medium or changing the other culture conditions the cells in these cultures could be induced stratification and terminal differentiation. The results suggest that the growth, proliferation and differentiation of cultured human epidermal keratinocytes can be controlled and regulated someway.

Objective To investigate osteocyte density as a potential index of bone biomechanical property. Methods Forty 7-month-old female Sprague-Dawley rats were randomly group (EST) and sham operation group (SHAM). At 15 weeks postoperation, the compression test was performed on L5 vertebral body and micro-computed tomography (μ-CT) was used to estimate the three-dimensional bone mineral density (BMD) and three-dimensional microstructure parameters of L6 vertebral body. After fatigue damage testing, the L6 vertebral body was bulk-stained in 1% basic fuchsin and embedded in methylmethacrylate. Mounted bone slices were used to measure microcrack parameters and osteocyte density. Results At 15 weeks postoperation, osteocyte density (Ot. N/T. area) was significantly decreased in OVX group compared with SHAM group and EST group [(1268. 1 ±191.2)/mm2 vs. (1760. 8 ± 376.6)/mm2 and (1550. 9± 202.2)/mm2, F = 3.513,P<0. 05]. Maximum load (ML) was significantly decreased and the length of microcrack (Cr. Le) was significantly increased in OVX group compared with SHAM group, EST group and GEN group [(84. 4±16.9)N vs. (110.3±25.6),(103. 9±15. 8)and(110.1±4. 9)N; (58. 1±6.8) μm vs. (24.2±8. 1), (36. 5±9. 7)and(28.5±7. 5)μm, F=9. 561,3. 179, all P<0. 05]. Compared with SHAM group and EST group, bone trabecula connection density (Conn. D) was significantly decreased and trabecular separation (Tb. Sp) was significantly increased in OVX group [(47.4±7.4) m-3 vs. (71.8±16.0)and (74.0±12.7)m-3;(315.0±32.7)μm vs. (222. 5±21.7)and (273.3± 50.0)μm, F=7. 635,7. 007, all P<0. 05]. Bone mineral content (BMC) was lower in OVX group than that in SHAM group[(6.5±2. 2)g vs. (7. 9±1.2)g, P<0. 05]. When data in four groups were overall analyzed, Ot. N/T. Ar was positively correlated with ML, Conn. D and BMC (R2 = 0. 7874, 0. 1153, 0. 1309, all P<0. 05), but was negatively correlated with Cr. Le and Tb. Sp (R2 =0. 5738, 0. 3964, both P < 0.05). Conclusions Osteocyte plays a crucial role in maintaining bone biomechanical property and osteocyte density may be considered as a useful indicator for assessing bone biomechanical property.

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.

A rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometric ( RRLC-QTOF/MS) method was used to profile the metabolites of urine samples from atherosclerosis ( AS) patients and healthy controls and find the differential metabolites which could provide the scientific evidence to explain the pathogenesis and early disease diagnose. In the study, 15 AS patients ( age46. 84±2. 41 years) and 15 healthy controls ( age45 . 72±1 . 93 years ) was screened out by VaSera VS-1000 . The urine samples were analyzed by RRLC-QTOF/MS and the resulting data matrices were analyzed by multivariate statistical analysis ( Principal Component Analysis, PCA ) to find the potential biomarkers. The results showed that the urine samples of AS patients were successfully distinguished from those of healthy controls. Besides, a total of two significantly changed metabolites, uric acid and Guanidineacetic acid, had been found and identified as potential biomarkers, which suggested that the disorder of purine metabolism and amino acid metabolism played an important role in the mechanism of AS.

In this study, the rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry ( RRLC-QTOF/MS ) was used to profile the metabolites of urine samples from Childhood Pneumonia ( CP) patients and healthy controls and find the potential biomarkers which can support evidence to early diagnose and cure the disease. Choose 10 CP patients ( age 47. 72 ± 2. 35 months) and 10 healthy controls ( age 46 . 65 ± 1 . 97 months ) . The urine samples were analyzed by RRLC-QTOF/MS and then the resulting data matrices were analyzed by principal components analysis ( PCA ) to find the potential biomarkers. Urine samples of CP patients were successfully distinguished from those of healthy controls. A total of two significantly changed metabolites have been found and identified as potential biomarkers. It is suggested that the disorder of purine metabolism and amino acid metabolism may play an important role in the mechanism of CP.

A soft coral Sinularia sp., collected from the South China Sea, was selected to investigate the bioactive and chemical constituents. The EtOAc fraction were isolated by repeatedly silica gel and Sephadex LH-20 column chromatography to obtain lobophytolide A (1), 3-dehydroxylpresinularolide B (2), sarcophine (3), 3 beta-acetoxyisolobophytolide (4), Crassocolide D (5), (3E,7E,11E)-6-acetoxy-3,7,11,15(17)-cembratrien-16,14-olide (6). The structures of compounds 1-6 were determined on the basis of spectroscopic data analysis. All compounds were tested against a small panel of human tumor cell lines. And these compounds were obtained for the first time from this coral.
Animals ; Anthozoa ; chemistry ; Diterpenes ; chemistry ; isolation & purification ; Molecular Structure

Based on the chemical structures of magnolol and honokiol,a series of small molecular derivatives were designed for the treatment of Alzheimer's disease.Through the Discovery Studio,five compounds (6a-6e) exhibited the inhibitory activity against Aβ and Tau proteins in all of the designed compounds.Then the five compounds are chemically synthesized and their biological activities were tested by thioflavin T.The result showed that compound 6a had inhibitory effect on the aggregation of two kinds of target proteins at the concentration of 100 μmol/L,which deserves further research.

BACKGROUNDTo investigate the expression level and its clinical significance of the multi-drug resistance (MDR) associated protein in lung cancer specimens.

METHODSThe expression levels of Pgp, MRP, GST-π and TopoII of MDR associated protein were detected in 60 lung cancer samples and 30 paracancerous tissues by S-P immunohistochemistry method.

RESULTSThe positive rate of Pgp, MRP, GST-π and Topo II in the lung cancer tissues was 40.0% (24/60), 61.7% (37/60), 45.0% (27/60) and 81.7% (49/60) respectively, which was significantly higher than those in paracancerous tissues (P < 0.01). No significant correlation was observed among the expression of multi-drug resistance-related protein and TNM staging, pathological type, cell differention and lymph node metastasis status (P > 0.05).

CONCLUSIONSLung cancer has overexpression of multi-drug resistant-related genes during the oncogenesis. Combined detection of these genes may be useful for guiding the lung cancer chemotherapy.

Objective To investigate the molecular characteristic of fluoroquinolone resistant Streptococcus agalactiae (GBS) isolates in Suzhou.Methods Totally 46 fluoroquinolone resistant GBS strains were collected,and then subjected into PCR and two resistance genes were sequenced and their high frequency mutation sites,gyrA and parC were analysed.Results Among these isolates,the most frequent gyrA mutation was gyrA_S81L (TCA→TTA),which displayed 93.4％ (43/46) with substituted at No.81 site of amino acid from Ser to Leu.However,the most frequent parC mutation was mutated at No.79 site of amino acid from Ser to Tyr orPhe,with two mutations parC_S79Y (73.9％,34/46) and parC_S79F(17.4％,8/46).The analysis showed that the major mutation patterns are gyrA_S81L with parC_S79Y (73.9％,34/46),gyrA_S81L with parC(13.0％,6/46).Conclusion The most frequent mutations of fluoroquinolone resistant gene in Suzhou are gyrA and parC and the major mutation patterns are gyrA_S81L/parC_S79Y and gyrA_S81L/parC_S79F.