Objective To study the effects of imatinib mesylate on the apoptosis in human melanoma cell line M14.Methods M14 cells were cultured in vitro in the presence of imatinib mesylate at three concentrations(5,10 and 20 μmol/L)for 96 hours.Sebsequeutly,annexin V-FITC and propidium iodide(PI)double staining flow cytometry and terminal deoxvnucleotidvl transferase (TdT)-mediated dUTP nick end labeling (TUNEL)were used to detect the cell cycle and apoptosis,respectively,DAPI staining to observe the mot-phological changes.and Western blot to measure the protein expressions of bcl-2 and bax in cells.Results Imatinib mesylate of the three concentrations could induce an evident increase in the apoptosis in M14 cells.Compared with untreated M14 cells,an increase of cell population in S phase was observed in imatinib mesylate.treated cells(P<0.05),along with a decline in cell population in G2/M phases(P<0.01).Annexin V/PI double staining and TUNEL revealed a significant increase in the rate of early apoptosis and in the acount of apoptotic cells,respectively,in M14 cells treated with imatinib mesylate of the three concentrations(all P<0.01).After treated with imatinib mesylate of 20 μmol/L.there was a morphological change characteristic of apoptosis in M14 cells,together with an upregulated expression of bcl-2(t=15.46,P<0.01)and downregu-lated expression of bax(t=25.53,P<0.01).Conclusions Imatinib mesylate can interfere with the process of cell cycle of and induce the apoptosis in M14 cells,which may be mediated through mitochondrial pathway.

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.

Appropriate selection and measurement of lead biomarkers of exposure are critically important for health care management purposes, public health decision making, and primary prevention synthesis. Lead is one of the neurotoxicants that seems to be involved in the etiology of psychologies. Biomarkers are generally classified into three groups: biomarkers of exposure, effect, and susceptibility.The main body compartments that store lead are the blood, soft tissues, and bone; the half-life of lead in these tissues is measured in weeks for blood, months for soft tissues, and years for bone. Within the brain, lead-induced damage in the prefrontal cerebral cortex, hippocampus, and cerebellum can lead to a variety of neurological disorders, such as brain damage, mental retardation, behavioral problems, nerve damage, and possibly Alzheimer's disease, Parkinsons disease, and schizophrenia. This paper presents an overview of biomarkers of lead exposure and discusses the neurotoxic effects of lead with regard to children and adults.
Alzheimer Disease ; chemically induced ; metabolism ; pathology ; physiopathology ; psychology ; Animals ; Behavior ; drug effects ; Biomarkers ; metabolism ; Brain ; metabolism ; pathology ; physiopathology ; Brain Diseases ; chemically induced ; pathology ; physiopathology ; Environmental Exposure ; adverse effects ; Humans ; Lead ; pharmacokinetics ; toxicity ; Lead Poisoning ; etiology ; metabolism ; pathology ; physiopathology ; psychology ; Neurotoxicity Syndromes ; etiology ; metabolism ; pathology ; physiopathology ; psychology ; Parkinson Disease, Secondary ; chemically induced ; metabolism ; pathology ; physiopathology ; psychology ; Schizophrenia ; chemically induced ; metabolism ; pathology ; physiopathology

Objective To explore the clinicopathologic features of pityriasis lichenoides et varioliformis acuta (PLEVA).Methods A retrospective analysis was performed.Clinical and histological data were collected from 60 patients with PLEVA.The clinicopathologic features of PLEVA were analyzed.Results Among the 60 patients with PLEVA,32 (53.3％) were aged 2-18 years,and 28 (46.7％) aged 19-49 years.Skin lesions were distributed in a diffuse pattern in 50 (83.3％) patients,in a central pattern in 2 (3.3％) patients,and in a peripheral pattern in 8 (13.4％) patients.Nineteen (31.6％) patients had a history of upper respiratory infection.Histopathological examination revealed liquefactive degeneration of basal cells and perivasculitis in the dermis in all the 60 cases,neutrophil abscess formation in the stratum corneum in 26 (43.3％) cases,keratinocyte necrosis in the epidermis in 41 (68.3％) cases,generalized liquefactive degeneration in 30 (50.0％) cases,migration of lymphocytes into the epidermis in 43 (71.6％) cases,Pautrier's microabscess formation in 2 cases,varying degrees of extravasation of erythrocytes into the epidermis in 46 (76.7％) cases,fibrinoid necrosis of blood vessel walls in the dermis in 3 cases.PLEVA progressed into granuloma fungoides in 1 patient.Twenty patients underwent immunohistochemical examination,and 3 of them showed monoclonal hyperplasia of T cells.Conclusions PLEVA has characteristic clinical manifestations,and the combination of pathological and clinical examination is the gold standard for its diagnosis.

Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P＜0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P＜0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.

Objective To investigate the possible effect of citrate on electrolyte metabolism in healthy people with different genders and races and provide a reference for the possible clinical interventions.Methods A cross over,placebo-controlled study was conducted in 22 age-matched Chinese(11 males and 11 females)and 10 male Caucasian volunteers after informed consents were obtained.Volunteers received of saline solution,separated by a wash-out period of two to three weeks.Serial blood and urine samples were collected during the observation period and analyzed for the selective biochemical parameters.Results Comparable basal levels of serum albumin[male(43.05±1.81)g/L vs female(42.26±2.67)g/L]and serum ionized calcium[male(1.27±0.04)mmol/L vs female(1.26±0.04)mmol/L]were observed between different genders of Chinese volunteers.However,citrate intervention led to more pronounced decrease of ionized calcium level in Chinese females compared to Chinese males[-28.68%(-20.00%--35.2%)vs-23.84%(-16.53%--29.32%),t=3.19,P ＜ 0.01].There was no differences of the levels of serum inorganic phosphate[-18.81%(-3.16%--25.09%)vs-19.23%(-1.22%--32.16%),t=0.36,P＞0.05]and albumin[-0.32%(3.27%--7.60%)vs 1.88%(6.03%--9.31%),t=0.47,P＞0.05].Independent of gender,citrate intervention resulted in an increased excretion of urine calcium in Chinese volunteers[before 0.34(0.09-0.87)vs after 0.96(0.18-1.47),t=6.66,P ＜0.01].Compared to Caucasian males,Chinese males has a higher basal level of serum ionized calcium [(1.27±0.04)mmol/L vs(1.22±0.02)mmol/L,t=3.7,P ＜0.01]and larger amplitude basal rhythm in serum albumin level[-11.72%(-5.70%--14.21%)vs-1.74%(2.43%--7.68%),t=7.43,P ＜ 0.01].Application of citrate resulted in comparable changes of serum ionized calcium [-23.84%(-16.53%--29.32%)vs-21.95%(-18.31%--30.92%)],phosphate[-19.23%(4.65%--32.16%)vs-12.68%(0.68%--42.19%)],albumin[-0.32%(1.05%--7.60%)vs-1.39%(1.87%--7.26%)]and urine calcium excretion[237.70%(11.8%-935%)vs 234.37%(5.45%-504.00%)]between Chinese and Caucasian males(t=0.32,0.03,0.25 and 0.04 respectively,P＞0.05).Serum levels of magnesium were not influenced in all volunteers during two interventions.Conclusions Independent of race and gender,the invention of citrate results in comparable changes of serum magnesium,inorganic phosphate and albumin.The effect of citrate on ionized calcium levels between genders implicates a higher risk for hypocalcemic reactions in females compared to males undergoing automatic apheresis procedures.

AIM:To investigate the therapeutic and preventive effects of paeoniflorin ( PF) on APP/PS1 mice, and to explore the possible mechanism. METHODS:Fifteen male 5-month-old APP/PS1 non-dominant mice were chosen as normal control group, 15 male 5-month-old APP/PS1 double transgenic mice were used as model group, and 15 male 5-month-old APP/PS1 double transgenic mice treated with 5 mg/kg PF by intraperitoneal injection were allocated in administation group. The learning and memory ability of the mice in each group was detected by Morris water maze. The apoptosis was assessed by TUNEL fluorescence staining. The protein expression of PI3K, Akt, p-PI3K, p-Akt, caspase-3, caspase-9, Bcl-2 and Bax in cerebral cortex and hippocampus was detected by Western Blot. The protein expression levels and distribution of caspase-3 and caspase-9 were detected by immunohistochemistry. RESULTS:(1) Compared with nor-mal control group, the learning and memory ability declined in APP/PS1 model group. Compared with APP/PS1 model group, PF obviously improve the ability of learning and memory in mice. (2) Compared with normal control group, the ap-optosis of nerve cells in APP/PS1 model group significantly increased and distributed in wider areas, while that in PF group was reduced (P<0.05). (3) Compared with APP/PS1 model group, PF could significantly lower pro-apoptotic factors, caspase-3, caspase-9 and Bax (P<0.05), and increase the expression of anti-apoptotic factors, p-PI3K, p-Akt and Bcl-2 (P<0.05). CONCLUSION:PF can up-regulate the expression of Bcl-2 and down-regulate the expression levels of caspase-9, caspase-3 and Bax via the activation of PI3K/Akt pathway, thereby inhibiting the nerve cell apoptosis and pro-tecting the nerve cells, so as to treat neurodegenerative diseases.

The excipient processing is an essential part of traditional Chinese medicine processing, and understanding its scientific connotations is a critical scientific issue that urgently needs resolution. Building upon a foundation where the composition of traditional Chinese medicine substances is fundamentally clear, this paper applies the techniques and methods of chemoinformatics to the study of the excipient processing mechanism. Relevant information on traditional Chinese medicines processed with four kinds of excipients (wine, vinegar, salt and honey) was collected, including properties, taste, meridian tropism, chemical components, etc. Molecular descritors and skeletons corresponding to each chemical component were calculated using chemoinformatics to characterize the properties and structural features of the components. Characteristic components associated with the four excipients (wine, vinegar, salt and honey) were explored through multivariate statistical analysis and Murcko skeleton analysis. Further analysis, taking honey-processed Astragali Radix as an example, the focus was on the impact of the processing excipient honey on the solubility and permeability of its characteristic pharmacologically active components (isoflavones). It was discovered that the excipient honey can increase their solubility and permeability, emphasizing that the impact of processing excipients on the pharmacological classification properties is a key breakthrough in elucidating the mechanism of excipient processing. In summary, this study analyzed the characteristic components associated with the processing excipients "wine, vinegar, salt and honey" based on their composition characteristics, providing data support for the research on the mechanism of excipient processing from a biopharmaceutical perspective.

Appropriate selection and measurement of lead biomarkers of exposure are critically important for health care management purposes, public health decision making, and primary prevention synthesis. Lead is one of the neurotoxicants that seems to be involved in the etiology of psychologies. Biomarkers are generally classified into three groups:biomarkers of exposure, effect, and susceptibility.The main body compartments that store lead are the blood, soft tissues, and bone;the half-life of lead in these tissues is measured in weeks for blood, months for soft tissues, and years for bone. Within the brain, lead-induced damage in the prefrontal cerebral cortex, hippocampus, and cerebellum can lead to a variety of neurological disorders, such as brain damage, mental retardation, behavioral problems, nerve damage, and possibly Alzheimer’s disease, Parkinson’s disease, and schizophrenia. This paper presents an overview of biomarkers of lead exposure and discusses the neurotoxic effects of lead with regard to children and adults.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.