Objective To observe the expression of phosphoinositide 3-kinase(PI3K) and protein kinase B1 (Akt1) in PI3K/Akt signaling pathway in rat bones with fluorosis, and to reveal the mechanisms of the skeletal fluorosis. Methods Thirty-six SD rats were randomly divided into 3 groups (control group, low-dose fluorosis group, high-dose fluorosis group) and 12 rats were in each group according to body weight. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose groups were fed with water containing NaF 5.0,50.0 mg/L, respectively. Rats were sacrificed after 6 months of treating with fluoride and the serum was kept for testing the bone metabolic markers of none gla protein(BGP) and cathepsin K(Cath-K) by enzyme-linked immunosorbent assay(ELISA), the proteins and mRNA levels of PI3K and Akt1 in rat bones were detected by immunohistochemistry and real time PCR, respectively. Results Each group of serum BGP and Cath-k were compared, the difference was statistically significant(F = 73.45,39.36, all P < 0.05). The contents of BGP[(1.99 ± 0.62), (2.38 ± 0.16)μg/L] and Cath-K [(89.07 ± 19.66), (110.16 ± 9.81)pmol/L] in the low-and high-dose fluorosis groups were higher than those in the control group[(0.15 ± 0.03)μg/L,( 18.32 ± 2.27)pmol/L], and the high fluorosis group was obviously higher than the low fluorosis group (all P < 0.05). Each group of serum PI3K and Akt1 protein and mRNA were compared, the difference was statistically significant(F- 178.16,118.08,38.81,52.31, all P< 0.05). Compared to the control group (181.55 ± 4.24,188.46 ± 2.18,3.84 ± 1.69,4.33 ± 0.89), the protein and mRNA expressions of PI3K(171.66 ± 2.85,154.12 ± 4.15,11.31 ± 4.18,20.54 ± 6.68), Akt1(177.47 ± 3.16,156.42 ± 3.18,12.52 ± 3.13,19.43 ± 5.36) were higher in the low- and high-dose fluorosis groups (all P < 0.05), and the high fluorosis group was obviously higher than the low fluorosis group (all P < 0.05). Conclusions BGP and Cath-K contents could be used as bone metabolic indices in the endemic fluorosis disease. Fluoride can increase the expression of PI3K and Akt1 mRNA and protein in bone tissue of fluorosis rats, and PI3K/Akt1 signaling pathway may be involved in the pathogenesis of bone injury caused by fluoride.

ObjectiveTo investigate the expression of nuclear factor kappa B(NF-kB)-related mRNA and protein in bone tissue of rats with chronic fluorosis.MethodsThirty-six healthy SD rats,weighting 100 - 120 g,were randomly divided into three groups (twelve in each group ).Rats of control group were fed with tap water (NaF ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-dose group:5 mg/L,high-dose group:50mg/L) through drinking water.All rats were killed at the eight month and metaphysic of femoral was collected.Bone tissues were stained with hematoxylin-eosin and observed under optical microscope.Bone fluorine was detected by ashing-fluorin ion selective electrode method.Serum content of tartrate-resistant acid phosphatase 5b(TRACP 5b)was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of p50,p65 and IkBα's mRNA and protein in bone tissue was detected by real-time PCR and immunohistochemistry.ResultsBone sclerosis was observed under optical microscope.The contents of bone fluorine in both the low and high doses fluoride groups [(6.32 ± 1.23),( 10.89 ± 1.56) mg/kg] were significantly higher than that of the control [(3.06 ± 1.01 ) mg/kg,all P ＜ 0.05],and of that the high fluoride group was significantly higher than that of the low fluoride group(P ＜ 0.05).Serum content of TRACP 5b of the low fluoride group[(3.45 ± 1.85)U/L] was significantly higher than that of the control[(1.26 ± 0.23)U/L,P ＜ 0.05],but that of the high fluoride group[(2.74 ± 1.85)U/L] was lower than that of the low dose group(P ＜ 0.05).The mRNA expressions of p50 and IkBα in the low fluoride group(4.41 ± 0.44,1.15 ± 0.25) were significantly higher than that of the control(1.46 ± 0.10,0.26 ± 0.07,all P ＜ 0.05),but that of the high fluoride group(0.69 ± 0.09,0.14 ± 0.03) was lower than that of the low dose group(all P ＜ 0.05).The protein expressions of p50 and IkBα in the low fluoride group(152.96 ± 7.87,156.20 ± 9.75) were significantly higher than that of the control( 125.63 ± 9.85,118.97 ± 6.94,all P ＜ 0.05),but the high fluoride groups' ( 120.56 ±9.57,114.50 ± 7.61 ) was significantly lower than that of the low dose group(all P ＜ 0.05).ConclusionFluoride can lead to altered gene expression of NF-kB pathway,and the latter may be involved in fluoride induced bone damage.

Objective To investigate the relationship between change of relevant gene of nuclear factor kappa B(NF-κB) and osteoclast apoptosis in bone injury of rats with chronic fluorosis,and to reveal the mechanism of skeletal fluorosis.Methods Thirty-six healthy SD rats,weighting 100-120 g,were randomly divided into three groups(twelve in each group).Rats of control group were fed with tap water(NaF ＜ 1 mg/L) and the experimental rats were exposed to NaF(low-dose group:5 mg/L,high-dose group:50 mg/L) through drinking water to established chronic fluorosis model.All rats were killed at the eight month and metaphysic of femoral was collected.Bone tissues were stained with hematoxylin-eosin and observed under optical microscope.Serum content of tartrateresistant acid phosphatase 5b (TRACP 5b) was detected by enzyme-linked immunosorbent assay (ELISA).Osteoclast was identified and counted by tartrate-resistant acid phosphatase staining(TRAP).The expression of p50,IκBα,Bcl-2 and Bax's mRNA and protein of bone tissue was detected by Real-time PCR and immunohistochemistry.Results Bone sclerosis was observed under optical microscope.The content of TRACP 5b in serum and the number of osteoclast in the low fluoride group[(3.45 ± 1.85)U/L,(6.75 ± 1.29)/slice]were significantly higher than that of the control[(1.26 ± 0.23)U/L,(3.92 ± 1.38)/slice,all P ＜ 0.05],but that of the high fluoride group [(2.74 ± 1.85)U/L,(3.33 ± 1.07)/slice]were lower than that of the low dose group(all P ＜ 0.05).The mRNA expressions of p50,IκBα,Bcl-2 and Bax in low fluoride group(4.41 ± 0.44,1.15 ± 0.25,2.02 ± 0.11,1.25 ± 0.22) were significantly higher than that of the control(1.46 ± 0.10,0.26 ± 0.07,1.00 ± 0.06,0.74 ± 0.09,all P ＜ 0.05),but the high fluoride groups' (0.69 ± 0.09,0.14 ± 0.03,0.95 ± 0.08,0.62 ± 0.08) were lower than that of the low dose group(all P ＜ 0.05).The protein expressions of p50 and IκBα in the low fluoride group (152.96 ± 7.87,156.20 ± 9.75) were significantly higher than that of the control(125.63 ± 9.85,118.97 ± 6.94,all P ＜ 0.05),but the high fluoride group(120.56 ± 9.57,114.50 ± 7.61) were lower than the low dose group(all P ＜ 0.05).The protein expressions of Bcl-2 and Bax(170.61 ± 6.60,160.77 ± 7.66) and the ratio of Bcl-2/Bax (1.07 ± 0.08) were higher than the control(l10.73 ± 5.27,114.64 ± 5.83,0.96 ± 0.04,all P＜ 0.05),but the high fluoride group(81.70 ± 8.00,99.93 ± 3.83,0.81 ± 0.08) were lower than that of the control and the low dose group (all P ＜ 0.05).There was a significant positive correlation between protein expression of p50,IκBα and Bcl-2/Bax (r =0.587,0.676,all P ＜ 0.05).Conclusions Chronic fluorosis can cause change of the relevant gene of NF-κB in rat bone tissues and osteoclast apoptosis.The mechanism of skeletal fluorosis might be related to the abnormal of osteclast apoptosis caused by changes of NF-κB p50 and IκBα.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Abstract: The demand for reliable toxicological data of chemicals runs through every link of occupational health work. The prevention of occupational diseases involves high requirements for the standardization of chemical toxicity assessment in occupational health institutions. Good laboratory practice (GLP) emphasizes the integrity of the test process to trace and supervise the whole process of the test, which is conducive to the standardization of chemical toxicity identification. Therefore, the standardized construction of GLP laboratories is an important starting point for occupational health institutions to carry out chemical toxicity identification. In the construction and management process of GLP laboratories for chemical toxicity identification, occupational health institutions need to build a sound organization and operation system, carry out systematic training and assessment of personnel, establish standard operating norms and emphasize their importance, strengthen the management of facility environment and laboratory, pay attention to quality control and process supervision, and constantly improve their own ability level. To actively adapt to social development and market demand, to provide strong support for occupational health work.

OBJECTIVETo investigate the expressions of mRNA and protein of p38, Osx, PI3K, Akt1 in the rats bone with chronic fluorosis.

METHODSDental fluorosis were observed and the fluoride contents in the urine and bone were detected by fluorin-ion selective electrode. The morphologic changes and ultrastructure of rats' bone were observed by light and electronic microscopy. The expressions of protein and mRNA of p38, Osx, PI3K and Akt1 were detected by immunohistochemistry and real-time PCR, respectively. The contents of BALP and BGP in serum were detected by ELISA.

RESULTSThe rates of dental fluorosis in the fluorosis rats were increased, and the fluoride contents in bone and urine of the fluorosis rats were increased compared to the control group, the difference was statistically significant (P < 0.05). The bone trabeculae thickness and density and the thickness of bone cortex in fluorosis rats were remarkably increased, the space of bone trabeculae was reduced, and in accordance with the matching morphometrical indices, the difference was statistically significant (P < 0.05) as compared with the control rats. The contents of BALP [(54.61 ± 2.27) U/L] and BGP [(2.38 ± 0.16) µg/L]in the fluoride groups were higher than those in the control group, the difference was statistically significant (P < 0.05). Ultrastructurally, the broadening of the osseouslacuna was observed. The reduced protuberances of the osteocytes, the unclear organelle structure, pyknosis, karyotheca increasation and edged chromatin were also observed. Compared to the control group, the expressions of protein and its mRNA of p38, Osx, PI3K and Akt1 were higher in the fluorosis rats than those in the control rats, and the difference was statistically significant (P < 0.05). There is no any expression of p38, Osx, PI3K and Akt1 in the osteocytes in fluorosis rats.

CONCLUSIONSThe over-expression of p38, Osx, PI3K and Akt1 in bone tissue of fluorosis rats may relate to the accumulation of fluorine in the body. The bone injury mainly occur in the stage of the differentiation and proliferation. The upregulation of P38MARK signal path and PI3K/Akt1 signal path may be involved in the pathogenesis of bone injury caused by fluoride.

Alkaline Phosphatase ; blood ; Animals ; Bone and Bones ; metabolism ; pathology ; ultrastructure ; Fluoride Poisoning ; metabolism ; pathology ; Fluorides ; metabolism ; urine ; Fluorosis, Dental ; metabolism ; pathology ; Immunohistochemistry ; Microscopy, Electron, Transmission ; Osteocalcin ; blood ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Sodium Fluoride ; toxicity ; Transcription Factors ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

Objective To investigate the immediate therapeutic effect of medicated thread moxibustion on cervical spondylosis.Methods Seventy patients with cervical spondylosis meeting the inclusion criteria were randomly allocated to treatment and control groups by using random number table method, 35 cases each. The treatment group received medicated thread moxibustion and the control group, percutaneous superficial needling. The Visual Analogue Scale (VAS) score and the Clinical Assessment Scale for Cervical Spondylosis (CASCS) score were recorded in the two groups before treatment and at 30 min and 24 hrs after treatment. The clinical therapeutic effects were compared between the two groups. Results The VAS and the CASCS scores decreased significantly in the two groups at 30 min and 24 hrs after treatment compared with before treatment (P<0.05,P<0.01). There was no significantly statistical difference in comparing the VAS score between the two groups after the treatment (P>0.05). The CASCS score was lower in the treatment group than in the control group at 30 min after treatment (P<0.01) and lower in the control group than in the treatment group at 24 hrs after treatment (P<0.05). At 30 min after treatment, the clinical therapeutic effect in the treatment group was superior to that in the control group (P<0.01); at 24 hrs after the treatment, the clinical therapeutic effect in the control group was superior to that in the treatment group (P<0.05).Conclusions Medicated thread moxibustion has a definite immediate therapeutic effect on cervical spondylosis.

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

OBJECTIVETo discuss the reliability of SARS-CoV antibody detection for SARS diagnosis.

METHODSUsing SARS-CoV ELISA kit to detect relevant antibody in fresh serum of healthy, fever, probable, and suspect cases.

RESULTSThe positive rate is 0%, 40%, and 95% respectively in healthy, probable, and suspect cases.

CONCLUSIONSIt is reliable to detect SARS-CoV antibody in late suspect patients, but there will be high false-positive result in ordinary fever cases.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; blood ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Female ; Fever ; diagnosis ; Humans ; Male ; Middle Aged ; SARS Virus ; immunology ; isolation & purification ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology ; Time Factors

OBJECTIVETo establish molecular typing of Listeria monocytogenes isolates by pulsed-field gel electrophoresis (PFGE) for studying the epidemiologic characteristics of Listeria monocytogenes isolated from foodstuff in Guangdong province and to build up PFGE typing database of Listeria monocytogenes isolates for identifying the infectious resource of the outbreaks and other epidemiologic investigation.

METHODS"Standardized Protocol for Molecular Subtyping of Listeria monocytogenes by PFGE" was followed. BioNumerics software was applied on image analysis, database establishment, comparative and corresponding analysis.

RESULTS107 Listeria monocytogenes isolates were typed by PFGE, 41 PFGE types were observed among the isolates. The PFGE types were dispersive among these isolates. Listeria monocytogenes isolates were most frequently isolated in raw chicken while the most PFGE types were found in this type of food. The positive rate was relatively high in cold and iced foods. Only 1-2 DNA fragment difference occurred in 26 Listeria monocytogenes isolates by PFGE, so high degree of relatedness remained among these isolates. There were unique PFGE patterns in the regions of Shaoguan and Huizhou. From time to time, a number of isolates remained close relationship.

CONCLUSIONPFGE typing of the 107 Guangdong Listeria monocytogenes isolates demonstrated relative genetic diversity but a number of the isolates showed close relatedness.

Electrophoresis, Gel, Pulsed-Field ; methods ; Food Microbiology ; Genetic Variation ; genetics ; Genotype ; Listeria monocytogenes ; classification ; genetics ; isolation & purification ; Phylogeny