Objective To investigate the effects of oxidative stress and lipoic acid(antioxidant)on bone metabolism and explore the underlying mechanism. Methods A total of 24 Wistar rats aged 8 weeks were randomly divided into three groups. Osteoporosis rats model was established by bilateral ovaries deleted. Rat in lipoic acid group was injected with lipoic acid(60 mg/kg)for 8 weeks. The bone mineral density(BMD),steo?calcin,ALP,Ca,P,MDA,SOD and GSH?Px were detected. The levels of OPG and RANKL in serum were measured by Western blotting. OPG and RANKL mRNA were detected by real?time PCR. Results The level of BMD level in blood,SOD,GSH?Px,OPG mRNA and protein level in femur of osteoporosis group were significantly lower than the control group(P<0.05). On the other hand,steocalcin,ALP,MDA,RANKL mRNA and protein level were significantly higher than the control group(P<0.05). The level of BMD level in blood,SOD,GSH?Px,OPG mRNA and protein level of lipoic acid group were significantly higher than the osteoporosis group(P<0.05). The steocalcin,ALP,MDA,RANKL mRNA and protein level were significantly lower than the osteoporosis group(P<0.05). Conclusion Oxidative stress may increase osteoporosis through the upregulation of OPG/RANKL pathway in rats ,and antioxidant lipoic acid can alleviate the progress of osteoporosis.

Through cluster multistage sampling,a resident group of 6 412 subjects with hypertension in the rural area of Liaoning province were recruited.According to IDF and NCEP-ATPm criteria the age-standardized prevalences of metabolic syndrome were 36.2%and 25.9%respectively.The prevalence of metabolic syndrome decreased with advancing age,but increased with rising of blood pressure.

Voltage-dependent potassium channels (Kv) are involved in proliferation and transformation in mammary epithelial cells. In previous studies, several groups have detected various potassium channels in breast cancer cells, and they assumed that potassium channels are related to the development of breast carcinoma, although the precise mechanisms are still unknown. We have previously reported that 4-aminopyridine (4-AP), one kind of potassium channel (K(+) channel) blocker, could affect the proliferation of MCF10A cells. The aim of the present study is to explore the expression and properties of K(+) channels in human mammary epithelial cells (MCF10A) and whether Kv channels are required for the proliferation of MCF10A cell. Electrophysiological, MTT analysis, PCR and Western blot methods were used to identify a K(+) conductance which is involved in tumorigenesis and not yet be described in MCF10A cells. A voltage-dependent, outward rectification and 4-AP-sensitive K(+) current was observed in these cells. The perfusion of 5 mmol/L 4-AP significantly decreased the amplitude of Kv current from (912.5+/-0.6) pA to (275+/-0.8) pA (n=5, P<0.01), when cells were recorded using 800 ms voltage steps from a holding potential of -60 mV to voltage ranging from -60 mV to +60 mV. PCR analysis demonstrated that Kv1.1, Kv1.2, Kv1.3, and Kv1.5 were all expressed in MCF10A and MCF7 cells. Furthermore, the expression of Kv1.5 was much higher in MCF10A than that in MCF7. Inhibitory effect of 4-AP on cell proliferation was dosage-dependent. Incubation with 5 mmol/L 4-AP reduced MCF10A cell proliferation to 25.29% in 48 h. Western blot analysis showed the activation of ERK1/2 which related to cell proliferation was enhanced, while p38 activation was decreased by 4-AP treatment for 10 min. These data provided the first evidence of the Kv channels expression in MCF10A cell and 4-AP could inhibit the proliferation of MCF10A through blocking the potassium channels, and the mechanism may be related to regulating the activity of different members of cell proliferation signaling pathway of MEK/ERK.
4-Aminopyridine ; pharmacology ; Cell Line ; Cell Proliferation ; Epithelial Cells ; physiology ; Humans ; Potassium Channel Blockers ; pharmacology ; Potassium Channels, Voltage-Gated ; physiology

BACKGROUNDThe bacterial composition of periapical lesions in deciduous teeth has not been well documented. This study was designed to explore the bacterial compositions, especially the dominant bacteria in periapical lesions using 16S rRNA sequencing.

METHODSTissue samples were collected from 11 periapical lesions in deciduous teeth with primary endodontic infections. DNA was extracted from each sample and analyzed using 16S rRNA cloning and sequencing for the identification of bacteria.

RESULTSAll DNA samples were positive for 16S rRNA gene PCR. One hundred and fifty-one phylotypes from 810 clones were identified to eight phyla, and each sample contained an average of 25.9 phylotypes. In addition, 59 phylotypes were detected in more than two samples, and Fusobacterium (F.) nucleatum (8/11), Dialister (D.) invisus (8/11), Campylobacter (C.) gracilis (7/11), Escherichia (E.) coli DH1 (6/11), Aggregatibacter (A.) segnis (6/11), and Streptococcus (S.) mitis (6/11) were the most prevalent species. Furthermore, 45 as-yet-uncultivated phylotypes were also identified.

CONCLUSIONSChronic periapical lesions in deciduous teeth contained polymicrobial infections. F. nucleatum, D. invisus, C. gracilis, E. coli DH1, A. segnis, and S. mitis were the most prevalent species detected by 16S rRNA sequencing.

Bacteria ; classification ; genetics ; isolation & purification ; Bacterial Infections ; microbiology ; Child ; Child, Preschool ; Female ; Humans ; Male ; Periapical Tissue ; microbiology ; RNA, Ribosomal, 16S ; genetics ; Tooth, Deciduous ; microbiology

Endometriosis is a prevalent chronic disease that affects approximately 6% to 10% of reproductive-aged women. Although numerous researchers have endeavored to explore the etiology of endometriosis over a century, its etiology still remains an enigma. The exploration of pathophysiologic mechanism and novel therapy for endometriosis depends on ideal endometriotic models. In the previous decade, various endometriotic models have been established; therefore, we made a conclusion for available information on these models. This review summarized the common experimental models used in endometriotic studies, including their origins, characteristics, applications, and limitations. Endometriotic models played an important role in studying etiologies and novel treatments of endometriosis during the last decades. Among them, animal models and endometriotic cell lines were viewed as most common studying tools to explore the intrinsic entities of endometriosis. In addition, endometrial organoid also emerged and was regarded as an ideal studying tool for endometriosis research. Different research models collectively complement each other to advance the endometriosis research. The successful establishment of endometrial organoids means that organoids are expected to become an ideal model for studying endometriosis in the future.

The conversion of exogenous p-hydroxybenzaldehyde to p-hydroxy-methyl-phenol-beta-D-glucoside (gastrodin) was studied by using cell suspension culture of Datura tatula L. The chemical structure of this synthesized gastrodin was identified based on the spectral analysis and chemical evidence. The conversion procedure of p-hydroxybenzaldehyde into gastrodin by D. tatula L. cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the ferment liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of D. tatula L. was also isolated and identified. The efficiency of glucosylation of p-hydroxybenzaldehyde was remarkably enhanced by adding salicylic acid (0.1 mg/L) and keeping the lower pressure (0.001MPa) in 25L airlift loop bioreactor. The biotransformation of exogenous p-hydroxybenzaldehyde to gastrodin by cell suspension culture of D. tatula L. is a promising approach.
Benzaldehydes ; metabolism ; Benzyl Alcohols ; chemistry ; Bioreactors ; Biotransformation ; Datura ; metabolism ; Glucosides ; biosynthesis ; chemistry ; Salicylic Acid ; pharmacology ; Suspensions

AIM:To observe the changes of autophagy-related indexes during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT) and its effect on apoptosis in human normal hepatocytes.METHODS:LO2 cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS.The expression of glucose-regulated protein 78 (GRP78) , protein kinase R-like endoplasmic reticulum kinase (PERK) , activating transcription factor 4 (ATF4) , C/EBP homologous protein (CHOP) , autophagy-related gene 12 (Atg12) , autophagy-related gene 5 (Atg5) and microtubule-associated protein 1 light chain 3 (LC3) at mRNA and protein levels was determined by real-time PCR and Western blot.The apoptosis was analyzed by flow cytometry.The formation of autophagosomes was observed under transmission electron microscope.After the LO2 cells were pretreated with rapamycin at 400 nmol/L for 1 h and treated with DTT at 2.0mmol/L for 24 h, the effect of rapamycin pretreatment on the apoptosis was analyzed by flow cytometry.RESULTS:After treatment with DTT at 2.0 mmol/L for 6, 12 and 24 h, the mRNA and protein levels of GRP78, PERK, ATF4, CHOP, Atg12, Atg5 and LC3 in the LO2 cells were significantly higher than those in 0 h group (P<0.05).At the same time, the ratio of LC3Ⅱ/LC3Ⅰwas also increased after DTT treatment (P<0.05).Observation under transmission electron microscope showed that autophagosomes were found in the LO2 cells treated with DTT for 6, 12 and 24 h.After DTT treatment for 6, 12 and 24 h, the apoptosis rate of LO2 cells was significantly higher than that in DTT 0 h group, while the apoptosis induced by DTT was significantly decreased after rapamycin pretreatment (P<0.05).CONCLUSION:ERS induces autophagy and rapamycin pretreatment alleviates the apoptosis of LO2 cells to some extent.

AIM:To investigate the effect of SET7/9 (SET domain containing 7/9) -mediated endoplasmic reticulum stress (ERS) on protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway, and to explore the mechanisms of arsenic-induced hepatocyte apoptosis.METHODS:Human liver LO2 cells were divided into control group, arsenic poisoning model group, negative transfection group and SET7/9 siRNA transfection group.The apoptosis of the LO2 cells in each group was analyzed by flow cytometry.The protein levels of SET7/9, glucose-regulated protein 78 (GRP78) , PERK and p-PERK in the LO2 cells of each group were observed by Western blot.RESULTS:Inhibition of SET7/9 expression reduced the apoptotic rate of arsenic-induced LO2 cells.Arsenic exposure increased the expression of SET7/9 in the LO2 cells.Arsenic exposure increased the protein levels of GRP78 and p-PERK in the LO2 cells, but decreased the protein levels of GRP78 and p-PERK after transfection with SET7/9 siRNA (P<0.05).CONCLUSION:Arsenic exposure induces hepatocyte apoptosis by increasing SET7/9 to activate ERS by PERK signaling pathway.

OBJECTIVETo investigate the inhibitory effects of tyroservatide and its amino acid mixture on growth of hepatocarcinoma.

METHODSHepatocarcinoma in nude mice was induced by implantation of cells of human hepatocarcinoma cell line BEL-7402. The inhibition of hepatocarcinoma growth was determined by calculating the tumor volume and measuring the tumor weight. The effects of tyroservatide on tumor cells in nude mice were assessed by immunohistochemical staining of proliferating cell nuclear antigen (PCNA), electron microscopic observation of ultrastructure, and apoptosis of tumor cells using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL).

RESULTSTyroservatide significantly inhibited the growth of human hepatocarcinoma in nude mice, with an inhibiting rate more than 60%. But the mixture of amino acid did not show a significant inhibitory effect on the tumor growth. Tyroservatide also induced apoptosis of tumor cells and decreased the expression of PCNA in tumor cells.

CONCLUSIONTyroservatide may significantly inhibit the growth of human hepatocarcinoma in nude mice by inducing apoptosis and inhibiting proliferation of tumor cells.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; prevention & control ; Liver Neoplasms, Experimental ; metabolism ; pathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oligopeptides ; pharmacology ; Proliferating Cell Nuclear Antigen ; metabolism ; Tumor Burden ; Xenograft Model Antitumor Assays

Aged ; Aged, 80 and over ; Endometriosis ; diagnosis ; pathology ; Female ; Humans ; Middle Aged ; Postmenopause ; physiology ; Retrospective Studies