Relevant literatures on the relationship betw een fluorosis and oxidative stress were reviewed.Based on most of the original papers published in recent years,we can see the increased free radicals and oxidative stress may occur in certain stage of fluoride intoxication,but confirmation of the causality between oxidative stress and fluoride-induced damages still remains much work to do.

Objective: To analyze the death among children under 5 years of age in Huzhou City, Zhejiang Province from 2012 and 2021, so as to provide insights into reduction of mortality among children. Methods: The mortality surveillance data among children under 5 years of age in Huzhou City from 2012 to 2021 were collected from Children Death Report Cards and Surveillance Report among Children under 5 Years of Age, including gender, place of residence, date of death and death diagnosis. The trends in mortality and cause of death were analyzed among children under 5 years of age in Huzhou City from 2012 to 2021. Results: A total of 1 262 deaths occurred among children under 5 years of age in Huzhou City from 2012 to 2021, with mean annual mortality of 4.39‰, and the mortality appeared a tendency towards a decline (χ2trend=132.695, P<0.001). A total of 899 infants died, with mean annual mortality of 3.13‰, and 363 children at ages of 1 to <5 years died, with mean annual mortality of 1.26‰. The mortality appeared a tendency towards a decline among both infants (χ2trend=117.778, P<0.001) and children at ages of 1 to <5 years (χ2trend=19.201, P<0.001). A total of 724 local children died, with mean annual mortality of 3.33‰, and there were 538 deaths among floating children, with mean annual mortality of 7.65‰. The mortality appeared a tendency towards a decline among both local (χ2trend=43.728, P<0.001) and floating children (χ2trend=94.038, P<0.001). The five most common causes of death included preterm birth or low birth weight (207 deaths, 16.40%), drowning (155 deaths, 12.28%), accidental asphyxia (138 deaths, 10.94%), other congenital abnormalities (126 deaths, 9.98%), and congenital heart diseases (113 deaths, 8.95%). Conclusions The mortality appeared a tendency towards a decline among children under 5 years of age in Huzhou City from 2012 to 2021, and preterm birth or low birth weight was the predominant cause of death.

AlM: To study the effects of different concentrations of Coix seed oil on human retinal capillary endothelial cells ( HRCECs ) proliferation and vascular endothelial growth factor ( VEGF) expression in high glucose environment.
METHODS: HRCECs extracted from human fresher eyeball and cultured in vitro, and ultimately used in the experiment were the growth of 3rd ~ 4th cells, the experimental were divided into blank control group, low glucose control group, high glucose control group, high glucose + ( 50ü L/mL, 100ü L/mL, 200ü L/mL ) different concentrations Coix seed oil group. Detecting the multiplication of HRCECs by MTT, the immunocytochemical method was employed to detect the each group HRCECs of VEGF expression.
RESULTS:MTT assay results showed that: different concentrations of coix seed oil acted at HRCECs for 48h, inhibition of cell proliferation was significant difference compared with high glucose control group (P<0. 05). Within 48h showed concentration dependence. There was no statistical difference between the low glucose group and high glucose control group (P>0. 05). lmmunocytochemical assay showed that:50ü L/mL, 100ü L/mL, 200ü L/mL Coix seed oil acted at HRCECs 48h, the expression of VEGF decreased significantly compared with the high glucose control group ( P< 0. 05 ), and in a dose- dependent manner. However, in high glucose control group, the expression of VEGF was obvious higher than that of low glucose control group (P<0. 05).
CONCLUSlON:Coix seed oil can inhibit the HRCECs proliferation and suppress the VEGF expression in high glucose environment.

The rat single-pass intestinal perfusion model was applied to study the effect of CYP3A and P-glycoprotein on the absorption of buagafuran in lumen of rats. Buagafuran concentrations in intestinal perfusate and blood in vena mesenterica collected at different time points after perfusion were determined by GC-MS. Permeability coefficient of buagafuran was calculated by the equation [P(lumen) = -(Q/2pirl)Ln(C(out)/C(in)) and P(blood) = (deltaM(B)/deltat)/(2pirl)]. The effects of troleandomycin (TAO, CYP3A inhibitor), cyclosporin A (CYP3A/p-glycoprotein inhibitor) on the absorption of buagafuran in lumen were observed. After rat single-pass intestinal perfusion, the cumulative amount of buagafuran in mesenteric vein of rat was 73.4, 82.9, and 98.3 pmol x cm(-2) and were increased 3.9, 4.6, and 5.6 fold by addition of inhibitor of P-gp (LSN335984), CYP3A (TAO) or P-gp and CYP3A (CsA), respectively. Moreover, the metabolized fraction of buagafuran was decreased by 12%, 11% and 21% with inhibitors. The results suggested that the poor bioavailability of buagafuran was mostly due to the interplay of P-gp and CYP3A on the absorption, transport and metabolism of buagafuran in intestine of rats.

A simple, rapid and sensitive method was developed for the quantification of tenofovir in plasma of Beagle dogs using HPLC-MS/MS analysis. The analytes tenofovir and internal standard (IS) adefovir were separated on a Zorbax SB-C18 column (3.5 microm, 100 mm x 2.1 mm, Agilent, USA) with mobile phase of methanol/water containing 0.3% formic acid using a gradient elution mode at a flow rate of 0.2 mL x min(-1). The plasma sample preparation was a simple deproteinization by the addition of 20% trichloroacetic acid followed by centrifugation. The detection was performed in positive selected reaction monitoring (SRM) mode with an electrospray ionization (ESI) source. The reactions monitored were m/z 288.1-176.2 for tenofovir and m/z 274.1-162.2 for adefovir (IS). Linear detection responses were obtained for tenofovir ranging from 10 to 5 000 ng x mL(-1). The intra- and inter-day precisions (RSD%) was no more than 6.3% with high recovery and good stability for the quantification, indicating the present method was specific, fast, accurate and reliable. The method was successfully applied to the pharmacokinetic study of two tenofovir agents. Tenofovir dipivoxil fumarate (BP0018, test agent) and tenofovir disoproxil fumarate (reference agent) were orally administrated to 8 Beagle dogs according to the 2 x 2 crossover design. Comparing with the reference agent, the longer MRT and t1/2 were obtained in the group of BP0018, while no significant difference was observed in AUC(0-t), AUC(0-infinity), C(max) and t(max) between them, suggesting that tenofovir dipivoxil fumarate was bioequivalent to the tenofovir disoproxil fumarate in Beagle dogs.

It is valuable to establish a chemical-pharmacokinetic (PK)-pharmacodynamics (PD) fingerprint of traditional Chinese medicine (TCM) for comprehensively understanding the TCM integrated conception and revealing the material foundation. The chemical, metabolic in vitro, and PK/PD in vivo fingerprints of Schisandra chinensis (SC) alcoholic extract were established and comparatively analyzed using HPLC-UV-MS method, rat liver microsomes in vitro and CCl4 intoxicated rats in vivo. Four known effective lignans, schisandrin, schisantherin A, deoxyschizandrin and gamma-schisandrin, were detected as the standard references in SC alcoholic extract with high concentration. SC alcoholic extract and four lignans when incubated with rat liver microsomes produced several metabolites in NAPDH-dependent manner. Chemical fingerprint of some components with bioactivities were also identified in PK and PD fingerprints in normal and ALI rats that explained the material foundation of SC alcoholic extract for multiple pharmacological effects. Schisandrin, schisantherin A, deoxyschizandrin and gamma-schisandrin could be considered as the "PK marker" of SC alcoholic extract or its relevant preparations, while two metabolites of the four lignans, 7, 8-dihydroxy-schizandrin and another one (M(W) 432), could be recognized as drug-metabolism (DM) Marker. This work provides experimental data for the further studies of metabolism or material foundation of SC components.

Objective To observe endoplagmic reticulum stress in bone tissue of fluomsis rats and further explore the pathogenesis of skeletal fluorosis.Methods 48 Wistar rats were divided into 4 groups according to their body mass.The control and low.calcium group were fed with normal diet(0.79%calcium)and low-calcium diet(0.79%calcium)respectively,and both drank tap water(sodium fluoride concentrations＜1 mg/L).High fluoride and low.calcium plus high-fluoride groups were fed with normal diet(0.79%calcium)and low-calcium diet (0.79%calcium)respectively,and both drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L).During experimental period,rats were measured body mass once a week with a stand diet and water available ad libiturn.The experimental period was 3 months.The biochemical techniques were used to test the indicators of oxidative stress and ALP in seFum of fluorosis rats.The total RNA was extracted from the one side of the femur,and the transcription level of Bip,Xbp1,CHOP and PDI were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results The level of MDA in serum of low-calcium plus high-fluoride group wag higher than that of the control[(14.74±3.11)μmol/L vs(10.15±1.96)μmol/L,P＜0.05];the activity of GPx was ma~edly higher in hish-fluoride group compared with the control[(3.87±0.41)×103 U/L vs(2.85± 0.55)×103 U/L,P＜0.05];the level of uric acid in sel'um was significantly lower both in high-fluoride group and low-calcium plus high-fluoride group compared with the respective control and the low-calcium group[(73.95± 9.52)μmol/L vs(110.43±25.48)μmol/L,(54.32±22.09)μmol/L vs(101.71±17.01)μmol/L,P＜0.05]. The activity of ALP wag obviously higher in low-calcium plus high-fluoride group compared with the control [(24.77±4.57)×103U/L vs (12.91±3.97)×103U/L,P＜0.01)].The mRNA expression of Bip/GAPDH in bone tissue was markedly higher in bone of high-fluoride group and low-calcium plus high-fluoride group compared with the control(1.38±0.24,1.35±0.12 vs 1.14±0.06,P ＜ 0.05). The expression of Xbp1/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control and the low-calcium group (1.48±0.20 vs 1.02±0.25,1.07±0.25,P ＜ 0.05 or ＜ 0.01);and CHOP/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control(0.84±0.18 vs 0.52±0.07,P ＜ 0.05 ). Conclusions Accelerated osteogenetic action is seen in fluorosis rats,accompanied by oxidative stress and bone endoplasmic reticulum stress,which is likely involved in the pathogenesis of skeletal fluorosis.

Objective To establish a nerve stem cell model possessing the potentiality of differentiating intoγ-aminobutyric acid neuron-like cells(GABAergic-like cells),and provide eligible investigative vector for study on GABAergic neurons degenerative diseases. Methods Dibutyryl cyclic adenosine monophosphate (dbcAMP)was applied to induce human neuroblastoma SH-SY5Y cells,and the SH-SY5Y cells were divided into control group (0 mmol·L-1 dbcAMP),0.3 mmol·L-1 dbcAMP group,0.6 mmol·L-1 dbcAMP group,1.0 mmol·L-1 dbcAMP group and 2.0 mmol·L-1 dbcAMP group;the morphological changes of SH-SY5Y cells were observed.The Image-Pro Plus 5.0 software was used to measure the length of neurites of the neuron-like cells, and the percentage of the SH-SY5Y cells with neurites longer than 30 μm was calculated. The immunofluorescence cytochemistry technique was utilized to test GAD65 positive cells,and the positive rate of immune response was calculated.Results The results of light microscope observation showed that the cells in control group were polygonal,circular or shuttle type with smooth membrane and clear boundaries. With the increasing of the concentrations of dbcAMP and the prolongation of time,the morphology of SH-SY5Y cells’bodies became smaller with longer processes in dbcAMP groups.The cells interwined each other and showed mature neuron phenotype in 1.0 mmol·L-1 dbcAMP group.Compared with control group(31.4%±4.2%),the percentages of the cell with neurites longer than 30 μm in 0.3, 0.6, 1.0, and 2.0 dbcAMP groups (40.1%± 5.7%, 47.5%± 6.2%, 73.1%±3.2%,and 74.3%± 6.1%)72 h after induction were significantly increased(P<0.05 or P<0.01). Compared with control group (10.2%± 2.1%), the GAD65 positive expression rates in 0.3, 0.6, 1.0,and 2.0 mmol·L-1,dbcAMP groups(22.1%±2.4%,46.9%±3.2%,70.7%±3.4%,and 72.3%±3.7%)72 h after induction were significantly increased(P<0.05 or P<0.01).Conclusion The SH-SY5Y cells have the potentiality of differentiating into GABAergic-like cells, and 1.0 mmol · L-1 is the optimal concentration of dbcAMP.

Objective To study the expression of skin aspartic protease (SASPase) in lesions of cutaneous lupus erythematosus (CLE) and its role in the pathogenesis of CLE.Methods Skin samples were resected from the lesions and normal skin of 9 patients with CLE,including 3 cases of subacute cutaneous lupus erythematosus (SCLE),4 cases of discoid lupus erythematosus (DLE) and 2 cases of acute cutaneous lupus erythematosus (ACLE).Keratinocytes were isolated from the tissue samples and cultured in serum-free medium.Total proteins were extracted from the keratinocytes and separated by two-dimensional gel electrophoresis.ImageMaster 2D analysis software was used to assess differentially expressed proteins in keratinocytes between the lesional and normal skin,which were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).The expression levels of SASPase were further determined by Western blot.The data were analyzed statistically by Student's t test.Results Keratinocytes were isolated from the tissue samples and successfully cultured in vitro.Two-dimensional electrophoresis profiles of proteins from the keratinocytes were obtained with high resolution and reproducibility,and the average matching protein spots were about 1200 with the matching rate higher than 80％.As Western blot showed,the relative expression level of SASPase was 0.463 ± 0.018 in keratinocytes from the lesional skin,and 0.145 ± 0.011 in those from the normal skin (P ＜ 0.05).The Western blot results were consistent with those of two-dimensional electrophoresis.Conclusion The initiation and progression of CLE seem to be associated with the abnormal activation and overexpression of SASPase.

ObjectiveTo study central venous oxygen saturation (ScyO2) in controlled hemorrhagic shock rabbits resuscitation process as a transfusion trigger and traditional transfusion trigger of comparison.MethodsSelection New Zealand pure line of rabbit 32 only,simple randonly divided into 4 groups,groups A and B for the observation group,groups C and D as control group,groups of eight only.A,B,C,D four groups respectively by ScvO2 ≤70％,ScvO2 ≤75％,hemoglobin (Hb)≥8g/dl,blood loss for the whole blood volume≥30％ as transfusion trigger.From right femoral artery bloodletting 10 minute inside,made the MAP to about (40 ± 5 )mmHg,and maintained the blood pressure 60 minutes,established controlled hemorrhagic shock rabbits of animal model.And then started to resuscitate,with colloid and crystalloid infusion according to the proportion 1∶2,infusion rate of about 10 ～ 15ml/( kg · h),according to the blood pressure and heart rate,and proper adjustment according to the different requirements of each group conducted a blood transfusion.Monitoring based value,shock,shock treatment 30 minutes,60 minutes,120 minutes,180 minutes all time points,and various indexes of blood loss,blood transfusions,crystalloid and colloid fluid volume and so on.ResultsIn shock treatment observation group A late blood pressure,pH,BE,HCO3-,O2ER etc compared with the other three groups had obvious statistical differences ( P ＜ 0.05 ),group B with C and D two groups at the same time points each monitoring were no significant differences ( P ＞0.05 ).The volume of transfusion group C was most,compared with the other three groups were significant difference ( P ＜ 0.05 ),group D of blood transfusions than A,B two groups (P ＜ 0.05 ),groups A and B infused colloid fluid,crystal fluid volume than groups C and D ( P ＜ 0.05 ),each group blood lossed without significant difference.ConclusionScvO2 for controlled hemorrhagic shock rabbit resuscitation monitoring can guide controlled hemorrhagic shock rabbit of blood transfusions,according to ScvO2 ≤75％ transfusion with traditional according to Hb or blood loss transfusion trigger comparison,can achieve the same resuscitation effect,and can more accurately and individualized guide transfusion,reduce unnecessary blood transfusions,save resources.