Objective To investigate the possible pathological role of mitochondrial apoptosis pathways and its factors in fluorosis-induced apoptosis of human hepatocellular carcinoma cell strain (HepG2).Methods Under the stimulation of 1,3,6 and 9 mmol/L concentrations of NaF in vitro for 24 h (n =5),while normal control group was cultured under normal condition,the cytotoxicity was measured with MTT.The mitochondrial apoptosis inducing factor (AIF) was measured at both mRNA (n =5) and protein levels (n =6),respectively,by real-time PCR and Western blotting.The mitochondrial apoptosis related factors,such as B-cells lymphoma-2 (Bcl-2),Bcl-associated X protein (Bax),cytochrome C,caspase-9 and caspase-3 were measured at protein levels (n =6).Results After treated with 0,1,3,6 and 9 mmol/L NaF for 24 h,the cell absorbance of HepG2 cells was 0.307 ± 0.031,0.333 ± 0.028,0.230 ± 0.011,0.178 ± 0.001 and 0.152 ± 0.003,respectively,and the differences were statistically significant among groups (F =82.224,P ＜ 0.01).After treated with 3 mol/L NaF for 24 h,the mRNA level of AIF was [(153.14 ± 5.41)％] which was increased compared to the control group [(100.00 ± 4.70)％,t =-4.73,P ＜0.05].Under the same condition,the protein levels of AIF,Bcl-2,cytochrome C in cytoplasm,caspase-9 and caspase-3 were (152.16 ± 47.30)％,(171.90 ± 51.52)％,(458.00 ± 19.48)％,(527.17 ± 200.67)％ and (432.70 ±64.27)％,which were increased compared to those of the control groups [(100.00 ± 48.86)％,(100.00 ± 34.44)％,(100.00 ± 116.59)％,(100.00 ± 19.58)％ and (100.00 ± 137.16)％,t =-3.80,-3.96,-15.76,-4.64,-5.06,all P ＜ 0.05],while the protein levels of Bax and cytochrome C in mitochondrion were (24.66 ± 26.04)％,(72.99 ±45.34)％,which were decreased compared to those of the control groups [(100.00 ± 44.01)％,(100.00 ± 34.14)％,t =6.35,0.68,all P ＜ 0.05].Conclusion The mitochondrial apoptosis pathway and related factors may be involved in NaF-induced cell death in HepG2 cells.

Objective To explore the influences of protein kinase Cβ (PKC3) inhibitor LY333531 on oxidative injury and apoptosis of SH-SY5Y cells induced by fluorosis.Methods The SH-SY5Y cell model of fluorosis was established,and the experiment was divided into three groups:control group [0.0 mmol/L sodium fluoride (NaF) and 0.0 μmol/L LY333531],the fluoride group (0.5 mmol/L NaF and 0.0 μmol/L LY333531),and the PKCβ inhibitor group (0.5 mmol/L NaF and 0.2 μmol/L LY333531),n =3.Flow cytometry was used to detect the changes of reactive oxygen species (ROS) and apoptosis rate,fluorescent probe technique was used to detect mitochondrial membrane potential after each group for 48 h.Results Compared with the control group [(3.32 ± 0.29) × 103,0.60 ± 0.09,(7.58 ± 1.20)％],the level of ROS [(5.99 ± 0.32) × 103] was increased,mitochondrial membrane potential (0.28 ± 0.06) was decreased,and the apoptosis rate [(18.00 ± 2.32)％] was increased in the fluoride group (all P ＜ 0.05);compared with the fluoride group,the level of ROS [(5.12 ± 0.25) × 103] was decreased,mitochondrial membrane potential (0.42 ± 0.03) was increased,and the apoptosis rate [(11.79 ± 0.70)％] was decreased in the PKCβ inhibitor group (all P ＜ 0.05).Conclusions Excess fluoride could cause oxidative damage and apoptosis in cells.PKC3 inhibitor LY333531 has a protective effect in oxidative damage and apoptosis by fluorosis.

Objective To investigate the influence of chronic fluorosis on protein kinase Cβ (PKCβ)/p66shc signal pathway in the brain of rats,and reveal the molecular mechanism of brain damage.Methods According to body weight by the random number table method thirty SD rats were divided into three groups of 10 each (half females and half males),the normal control group [less than 0.5 mg/L of fluorine (prepared with NaF) in drinking water],low fluoride exposure group (10.0 mg/L fluorine),and high fluoride exposure group (50.0 mg/L fluoride).The experiment period was 6 months.The protein level of PKCβ,p66shc,phospho-p66shc and preserved ammonia acyl isomerase (Pin1) in rat brain was detected by Western blotting.The level of neuron nuclear antigen (NeuN),p66shc and phospho-p66sh in brain of rats was detected by immunohistochemistry.Results By Western blotting,the levels of PKCβ,Pin1 and phospho-p66shc protein in brain tissue in high fluoride exposure group [(193.00 ± 57.53)％,(228.21 ± 71.14)％,(201.54 ±:50.86)％] were higher than those of the normal control groups [(100.00 ± 21.24)％,(100.00 ± 40.55)％,(100.00 ± 13.35)％,all P ＜ 0.05].By immunohistochemistry,the numbers of NeuN staining in brain tissue of the rats in both high and low fluoride exposure groups [(49.50 ± 12.57)％,(65.66 ±14.58)％] were lower than that of the control group [(100.00 ± 18.32)％,all P ＜ 0.01].The level of phospho-p66shc protein in brain tissue in high fluoride exposure group [(242.66 ± 93.01)％] was higher than those of the low fluoride exposure and the normal control groups [(152.53 ± 60.65)％,(100.00 ± 25.63)％,all P ＜ 0.01].Conclusion Chronic fluorosis has increased the expressions of PKCβ,Pin1 and phospho-p66shc at protein level in brain of rats,which may be related to the molecular mechanism of brain damage resulted from chronic fluorosis.

Objective: To investigate the effect of lovastatin on oxidative stress and apoptosis in neurons induced by β-amyloid peptide (Aβ). Methods: Primary culture of rat hippocampal neuron was treated with Aβ oligomers alone or combined with lovastatin. The levels of OH^-, H₂O₂, O₂·^-, malondialdehyde, superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities were measured by biochemical methods and protein expression of caspase-3 and bcl-2 was detected by Western blot. Results: As compared with the control group, treatment of 0.5 μmol/L Aβ oligomers for 48 h led to significant increase of OH^-, H₂O₂, O₂·^- and malondialdehyde content, inhibition of SOD and GSH-PX activities, enhanced caspase-3 expression and decreased bcl-2 expression. Interestingly, these neurotoxic modifications on the levels of OH^-, H₂O₂, O₂·^- and malondialdehyde content, SOD and GSH-PX activities, and the protein expression of cleaved caspase-3 and bcl-2 were significantly attenuated when the cells were pretreated with 0.1 μmol/L lovastatin for 24 h before exposure of Aβ oligomers. Conclusion Lovastatin may play an important role in antagonizing the neurotoxicity of Aβ through a mechanism likely related to the inhibition of oxidative stress.

Objective:To investigate the pathogenic factors of vulvovaginal candidiasis (VVC) in occupational women of childbearing age and the prognostic value of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin (IL)-2.Methods:184 patients from the Meishan Maternal and Child Health Hospital from June 2016 to June 2017 in the gynaecological clinic were selected for research.The self-made general situation questionnaire was used to investigate the situation of the patients, and the vaginal secretions of the patients were sampled for microscopic examination to diagnose vulvovaginal candidiasis. According to the prevalence and recurrence of VVC, the patients were divided into recurrent group (42 cases), non recurrent group (55 cases) and non-VVC group (87 cases). Single factor analysis and multi factor logistic analysis were used to analyze the pathogenic factors of VVC.Results:Among the 184 patients, 97 were diagnosed as VVC, with a 52.72% incidence rate. The results of single analysis showed that age, drinking sugary drinks, eating sweets, exercising, sedentary, emotional state, frequency of using pads in non menstrual period, wearing tights, history of vaginitis, frequency of vaginal washing, history of curettage, contraceptive method, first sexual intercourse age, number of sexual partners, whether washing vulva before and after sexual life were related to the incidence of vulvovaginal candidiasis in occupational women of childbearing age ( P<0.05). The results of multivariate analysis showed that sedentary, drinking sugary drinks, eating sweets, wearing tights, vaginal washing frequency, first sexual intercourse age, contraceptive method, number of sexual partners, history of curettage, history of vaginitis, and cleaning vulva before and after sexual life were all independent factors ( P<0.05). There was no significant difference in TNF-α level among the three groups ( P>0.05), while the IFN-γ and IL-2 levels were the lowest in the recurrent group, the second in the non recurrent group, and the highest in the control group ( P<0.05). Conclusions:The incidence rate of VVC in the women of childbearing age occupation is related to sedentary, sugar drinking, sweet food, tight pants frequency, vaginal irrigation frequency, first sex, contraceptive methods, sexual partners, curettage history, vaginitis history, sexual cleansing before and after sexual activity. Clinically, relevant factors can be intervened to reduce the incidence of the disease. The decrease of IFN-γ and IL-2 may increase the risk of VVC.

OBJECTIVETo observe the clinical efficacy differences between modified lateral position and prone position in patients treated with electroacupuncture (EA) for lumbar herniated disc (LHD).

METHODSSeventy-six patients with LHD were randomly divided into a lateral position group and a prone position group, 38 cases in each one. The acupoint selection and treatment method were identical in the two groups except the position. Mingmen (GV 4), Yaoyangguan (GV 3), Dachangshu (BL 25), Xiaochangshu (BL 27), Zhibian (BL 54) and Huantiao (GB 30) were selected. EA was given three times a week, ten times were taken as one course and totally 20 times were given. The visual analogue scale (VAS) and Japanese orthopaedic association (JOA) scale were taken as efficacy criteria, which were evaluated before and after treatment as well as one month after treatment.

RESULTSAfter treatment, VAS and JOA were significantly improved in the two groups (lateral position group:JOA 10.60±2.60 vs 18.92±3.87, VAS 8.13±0.99 vs 2.34±0.81; prone position group:JOA 10.94±2.06 vs 17.02±3.96, VAS 8.02±1.05 vs 2.86±0.96, all<0.01); the VAS and JOA in the lateral position group were higher than those in the prone position group (both<0.05). One month after treatment, VAS and JOA were significantly improved in the two groups (all<0.01), which was more significant in the lateral position group (both<0.05).

CONCLUSIONSThe treatment position could influence the efficacy of EA for LHD, and lateral position pre-sents certain advantages to prone position group.