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.

Actins ; analysis ; Antigens, CD34 ; analysis ; Carcinoma, Papillary ; classification ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Keratin-19 ; analysis ; Keratin-20 ; analysis ; Muscle, Smooth ; chemistry ; Pancreatic Neoplasms ; classification ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; analysis ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis

OBJECTIVETo study the effect of solid dispersion technology and inclusion technology on dissolution performance of Pulsatillae total saponins, and preliminarily investigate its mechanism.

METHODThe solid dispersion of Pulsatillae total saponins-PEG 4000 was prepared by the melting method. The inclusion compound of Pulsatillae total saponins-hydroxypropyl-beta-cyclodextrin ( HP-beta-CD) was prepared by the freeze-drying method. The properties of solid dispersion and inclusion compound were identified by using IR, DSC and NMR. And the dissolution of solid dispersion and inclusion compound were also determined by the small glass method.

RESULTIR, DSC and NMR results showed the formation of solid dispersion and inclusion compound. In terms of the dissolution, the inclusion compound ranked first, which was followed by solid dispersion and bulk pharmaceutical chemicals.

CONCLUSIONThe inclusion technology could significantly increase the dissolution of Pulsatillae total saponins, whereas the solid dispersion showed no notable solubilization effect.

Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Saponins ; chemistry ; Solubility ; Spectrophotometry, Infrared

The paper deals with a portable analyzing monitor system with liquid crystal display (LCD), which is low in power consumption and suitable for China's specific conditions. Apart from the development of the overall scheme of the system, the paper introduces the design of the hardware and the software. The 80196 single chip microcomputer is used as the central microprocessor to process and real-time electrocardiac signal data. The system have the following functions: five types of arrhythmia analysis, alarm, freeze, and record of automatic paperfeeding. The portable system can be operated by alternate-current (AC) or direct-current (DC). Its hardware circuit is simplified and its software structure is optimized. Multiple low power consumption and LCD unit are adopted in its modular designs.
Arrhythmias, Cardiac ; diagnosis ; Computers ; Electrocardiography, Ambulatory ; instrumentation ; Equipment Design ; Humans ; Microcomputers ; Software

To study the effect of Wansheng Huafeng Dan (WSHFD) and mercuric chloride on renal mercury (Hg) extraction transporters (Oat1, Oct2), renal mercury excretion transporters (Mrp4, Mate2K), renal mercury accumulation and kidney injury molecule-1 (Kim-1). The ancient prescription of WSHFD containing 10-fold Hg caused much lower renal mercury accumulation and renal toxicity than HgCl2 in rats, with less effect on renal transporters than HgCl2. The above indicators had no significant difference in WSHFDO, WSHFD2 and WSHFD3 groups, indicating no effect of WSHFD with reduced or no cinnabar.
Animals ; Ardisia ; chemistry ; Biological Transport ; drug effects ; Cell Adhesion Molecules ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Gene Expression ; drug effects ; Kidney ; drug effects ; metabolism ; Male ; Mercuric Chloride ; metabolism ; Multidrug Resistance-Associated Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVELate onset neonatal septicemia (systemic infection after 72 hours of life) remains a major cause of neonatal morbidity and mortality. Early treatment with appropriate antibiotics is critical since infected infants can deteriorate rapidly. The aim of this study was to review the pathogens responsible for late onset neonatal septicemia (LONS) and their antimicrobial susceptibilities in order to guide the initial selection of appropriate antibiotics for infants with suspected LONS.

METHODSA retrospective chart review of all cases with LONS seen in the neonatal intensive care unit (NICU) of Yuying Children's Hospital of Wenzhou Medical College from January 1, 2002 to December 31, 2005 was conducted. All cases were selected based on the clinical presentation and at least one positive result of blood culture. The basic clinical characteristics and the results of blood culture and antimicrobial susceptibilities were analyzed.

RESULTSA total of 102 cases with LONS were identified. Among those 102 cases, 80 were community acquired (infants admitted from home and the blood culture was done on admission) and 22 were hospital acquired (infants became sick while in the NICU and the blood culture was done prior to use of antibiotics). The clinical presentations were non-specific. Compared to the infants with community acquired LONS, infants with hospital acquired LONS were usually born more prematurely (mean gestational age 33 +/- 3 vs 39 +/- 2 wks, t = 2.255, P < 0.01), with lower weight (mean weight 1.79 +/- 0.70 vs 3.23 +/- 0.67 kg, t = 8.818, P < 0.01) and with younger age (mean age 12 +/- 6 vs 16 +/- 7 days, t = 7.581, P < 0.05). Of the 102 cases, a total of 103 strains of bacteria were isolated. Among the pathogenic bacteria isolated, the most common were coagulase-negative Staphylococcus (CoNS) (50/103, 48.5%), followed by Klebsiella pneumoniae (16/103, 15.5%). The main pathogens for community acquired LONS were Staphylococcus species and Escherichia coli. The most important pathogen responsible for hospital acquired LONS was Klebsiella pneumoniae. Most (> 80%) of the Staphylococcus especially CoNS were resistant to common antibiotics such as penicillin, erythromycin and cefazolin. Significant numbers (6/9) of Staphylococcus aureus isolated were methicillin-resistant Staphylococcus aureus (MRSA). However, all of the Staphyloccus isolates were sensitive to vancomycin. Almost all (15/16) of the Klebsiella pneumoniae isolated were multi-drug resistant due to production of extended-spectrum beta-lactamases (ESBLs). They were sensitive only to a few antibiotics such as carbapenems, aminoglycosides and quinolones. There was also one strain of vancomycin-resistant Enterococcus (VRE). Furthermore, there was no a single case of late onset neonatal sepsis due to infection with group B Streptococcus (GBS).

CONCLUSIONSThe clinical manifestations of late onset neonatal sepsis are usually non-specific. GBS is not a significant pathogen responsible for community acquired LONS in the Wenzhou area. There are increasing numbers of multi-drug resistant bacterial species isolated from the newborn infants with late onset neonatal septicemia, which is most likely due the non-restricted use of antibiotics in the hospitals as well as in the communities. A routine blood culture should be taken from any newborn infant who is suspected of LONS and empirical use of appropriate antibiotics should be initiated as soon as the blood specimen for culture has been drawn. To reduce the occurrence of multi-drug resistant bacteria, the use of antibiotics especially the third generation cephalosporins in neonates should be restricted as much as possible.

Community-Acquired Infections ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Drug Resistance, Multiple, Bacterial ; Female ; Humans ; Infant, Newborn ; Male ; Retrospective Studies ; Sepsis ; microbiology

Objective: To conduct an epidemiological investigation on a cluster of overseas imported cases of coronavirus disease 2019(COVID-19)in Qingtian County, Lishui,so as to provide reference for the prevention and control of COVID-19 imported from abroad. Methods: According to the COVID-19 Prevention and Control Program (FiFth Edition) issued by National Health Commission of China,a field investigation was employed and the close contacts of the case were tracked down ;the real-time reverse transcription polymerase chain reaction (RT-PCR) assay was performed to detect the nucleic acid of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from the sputum specimens or throat swabs of cases;the epidemiological characteristics,source of infection,route of transmission and disposal of the cluster were analyzed. Results: From March 1 to March 6,2020,eight confirmed cases and one asymptomatic case of COVID-19 were reported. Their median age was 33 years old. The nine cases all had no fever,no decrease in leukocyte and lymphocyte levels,and the clinical symptoms of seven cases were not obvious. The asymptomatic case had been tested negative for SARS-CoV-2 for six times,but had been weakly positive for IgM antibody and strongly positive for IgG antibody. Nine cases were all the employees of the same restaurant in Begamo,Italy. They lived in three independent villas and usually had lunch and dinner in the restaurant where they worked. Begamo had COVID-19 epidemic,but the staff of the restaurant did not take any protective measures such as wearing masks and environmental disinfection. Eight cases reported to have cold symptoms in Italy during mid February. Through a closed-loop management of“all people,all sites,all chains ”,such as treatment of current cases,screening and isolation of close contacts and health education,totally 372 close contacts were traced back,yet no fever or respiratory symptoms have been found. Conclusions It was a cluster of COVID-19 cases imported from abroad. The clinical symptoms of the cases were not obvious. Qingtian County adopted the closed-loop management in time,and effectively controlled the spread of the epidemic. No second-generation cases have been found.

OBJECTIVETo investigate the expression levels of nm23, P53, and S100A4 in gastric carcinoma and their relationships with clinicopathologic parameters and metastasis potential.

METHODSPathological specimens from gastric carcinoma,matched para-tumor tissues, metastatic lymph node and distant metastatic tissues were examined for the expression levels of nm23, P53, and S100A4 proteins by tissue microarray technique and immunohistochemistry.

RESULTSThe expression levels of P53 and S100A4 were upregulated (P< 0.01), while the expression of nm23 downregulated (P< 0.05) in gastric carcinoma compared with non-tumor tissues. S100A4 expression was significantly higher in distant metastatic tissues, while nm23 lower in metastatic lymph nodes than those in cancer tissues. Upregulating expression levels of nm23, P53, and S100A4 were significantly correlated with some malignant behaviour of gastric cancer. The expression rates of nm23+/P53+, P53+/S100A4+, and nm23+/S100A4+ immunohistochemical phenotypes were 48/74 (64.9%), 50/74 (67.6%), and 39/74 (52.7%). P53+/S100A4+, nm23+/S100A4+, and nm23+/P53+/S100A4+ phenotypes were associated with high metastasis potential of gastric cancer.

CONCLUSIONSAlteration of nm23, P53, and S100A4 expression may contribute to the development of gastric carcinoma. Nm23 and S100A4 proteins play a critical role in tumor metastasis. Co-detection of the expression of P53, nm23, and/or S100A4 can be used to evaluate high metastasis potential of gastric cancer.

Female ; Gene Expression ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; metabolism ; Neoplasm Invasiveness ; Neoplasm Staging ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism