Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endotoxemia ; blood ; drug therapy ; immunology ; Humans ; Lipopolysaccharide Receptors ; blood ; Lipopolysaccharides ; blood ; Phytotherapy ; Shock, Septic ; blood ; drug therapy ; immunology

Objective:To express and purify recombinant human collagen type Ⅲ and evaluate its properties.Methods:The recombinant genetic engineering strain pET30a(+)-1880/pACYCDuet-hy726/bL21(DE3) was constructed to stably co-express recombinant human type Ⅲ collagen (rhCol) and prolyl hydroxylase. rhCol was prepared and purified by E. coli high-density fermentation, salting out and column chromatography protein purification technology. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the purity of rhCol. The N-terminal amino acid sequences of rhCol were determined by automatic protein polypeptide sequencing instrument. The hydroxyproline content of rhCol was determined by ultraviolet spectrophotometry. The cellular compatibility of rhCol was evaluated by MTT assay. Results:The final wet weight of high-density fermentation was about 200 g/L. The expression level was about 3 g/L. The purity of rhCol by affinity chromatography was over 95%. The results showed that the hydroxyproline content of rhCol was 11.44%, and the rhCol products have good water solubility and cell compatibility.Conclusions:RhCol can be widely applied to the field of skin care and biomedicine as an excellent biological material.

Background and purpose: Multidrug resistance (MDR) is the dominating obstacle to the chemotherapy. There is strong evidence that the phosphoinositide 3-kinases (PI3Ks) signaling pathway is involved in MDR phenotype, however, the mechanism of MDR occurrence is still unknown. This study tended to investigate the regulating effect of PI3K/Akt signaling pathway and its downstream target genes in P-glycoprotein (P-gp) (ABCB1 gene encoding)-mediated MDR in human colon carcinoma HCT-116/L-OHP cells. Methods:Pretreatment with PI3K selective inhibitor LY294002 (20μmol/L) for 2 h, the sensitivity of L-OHP was evaluated by the CCK-8 (cell counting kit-8) assay in HCT-116/L-OHP cells, and the expressions of P-gp, LRP, MRP-2, Akt, p-Akt, IκB and p-IκB were evaluated by Western blot. The activity of ABCB1 promoter was evaluated by chromatin immunoprecipitation analysis (CHIP). Results: After inhibiting the activity of PI3K/Akt signaling pathway, the IC50 value of L-OHP decreased from(157.48±16.73)μg/mL to (53.68±3.18)μg/mL in HCT-116/L-OHP cells, and the reversal index was 2.93 (P<0.01). The expressions of P-gp, p-Akt and p-IκB were down-regulation compared with the concrol group (P<0.01), but the expressions of LRP, MRP-2, Akt and IκB didn't change signiifcantly. CHIP result has conifrmed that NF-κB protein could bind to the region of ABCB1 gene promoter in HCT116/L-OHP cells. Conclusion:Blocking of PI3K/Akt/NF-kB signal pathway could increase the drug sensitivity to MDR cells, inhibit the phosphorylation of p-Akt and p-IκB, and reversing ABCB1/P-glycoprotein-mediated multidrug resistance in colon carcinoma cells.

Objective To explore the primary teeth caries status of preschoolers in Pudong community and its relevant factors of parental knowledge-belief-practice (KBP).Methods Using stratified random cluster sampling,a total of 1 179 children from 9 kindergartens in Hudong community in Pudong new area were examined.Oral cavity check-ups and parental questionnaire survey were conduced to understand the primary teeth caries rate of preschoolers and its related influencing factors.Results The prevalence rate of primary teeth caries was 53.2％ and DMFT 2.79.The prevalence rate of primary teeth caries had statistical significance in different age groups (P ＜ 0.05).Multi-factor analysis showed that the influencing factors of primary teeth caries were age (P ＜ 0.05),parental education level (P ＜ 0.05),parental assistance of teeth brushing (P ＜ 0.05),children eating sweets before bedtime (P ＜ 0.05) and parental awareness for treating primary teeth (P ＜ 0.05).Conclusions The prevalence rate of primary teeth caries is high among the preschoolers of Pudong.And age,parental education level,parental preventive knowledge of teeth caries and poor eating habits are significant factors correlated with the incidence of dental caries in children.

Communicable Diseases ; diagnosis ; epidemiology ; Humans ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo explore the correlation between hand and face-mouth, so as to provide nerve reflex basis for the theory "Hegu (LI 4) regulates face and mouth".

METHODSSeven hundred and sixty-three participants who met the inclusive criteria were divided into different age groups. The skin around participants' thenar eminence was gently scraped to be observed whether there was an involuntary movement around the face or mouth, which was palmomental reflex. The results of palmomental reflex were recorded.

RESULTSThe total occurrence rate of palmomental reflex was 46.26%. For those who were 0 to 1 years old, the palmomental reflex was all positive; for those who were 21 to 36 years old, the positive rate was 20.45%, which was the lowest; for those who were 65 to 85 years old, more than half of them were positive. The majority of those who were 0 to 2 years old were bilateral positive palmomental reflex, while the majority of those who were 65 to 85 years old were unilateral positive palmomental reflex.

CONCLUSIONThere is a certain connection between hand and face-mouth. The occurrence rate of palmomental reflex changes from high to low over age increasing, and then changes from low to high with the aging, presenting a "high-low-high" U-shaped curve, which is possible related to the growth and recession of nervous system.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Face ; physiopathology ; Female ; Hand ; physiopathology ; Humans ; Infant ; Male ; Middle Aged ; Mouth ; physiopathology ; Reflex ; Young Adult

Seven emotions generation relates to the interaction between individual and external objective matters,which is accompanied with interaction between internal desire and external matters.The activity of seven emotions bases on essence of zang-fu organs,being regulated by five zang viscera.Heart is the upstream controller,liver is the key organ to maintain normal emotionds,spleen and stomach is the hub of emotional activities,lung is the auxiliary organ, kidney is origin of emotional generation.Five zang viscera's cooperation and interaction generate the seven emotions.

[Objective]To apply an acetonitrile-pretreatment method for proteomic analysis of serum of patient with hepatocellular carcinoma(HCC),searching for the proteins that might be related to occurrence and development of HCC.[Methods]Serum samples were recruited from 12 patients with primary HCC and 12 health adult.Firsdy,the sera were pretreated with acetonitrile (ACN),and the high-abundance proteins were removed,then two-dimensional electrophoreaia(2-DE)was performed to screen the differentially expressed proteins between the patient with HCC and health control.Finally,the differentially expressed proteins in the 2-DE gel were identified by a MALDI-TOF/TOF mass spectrometry.[Results]After the sera were pretreated with 0%,20% and 30% ACN,2-DE analysis showed that the number of the protein spots in the gels were 532±96,623±102,and 674±123,respectively.By the proteomic analysis of the ACN-pretreatment sera of patient with HCC and health control,we found that tetraiodo-L-thyronine and proapolipoproteinin were up-regulated,vitamin D-binding protein precurser and transferrin were downregulated in the HCC serum.[Conclusion]Proteomic analysis of serum using the ACN-pretreatment method can improve the detection of proteins which were non-specifically binding to the albumin.Abnormal expression of tetraiodo-L-thyronine,proapolipoproteinin,vitamin D-binding protein precursor and transferrin in the serum are related to HCC.

OBJECTIVETo evaluate the effect of immunoadsorption therapy in patients with myasthenia gravis (MG) and explore the mechanism.

METHODSThis investigation involved 20 patients with MG treated with immunoadsorption combined with hormonal therapy and another 20 with only hormonal therapy, and 15 healthy subjects served as the control group. Enzyme-linked immunosorbent assay (ELISA) was used to measure the changes in serum tumor necrosis factor-α (TNF-α) and interleukin-18 (IL-18) after the treatments, and the therapeutic effect of the treatments was evaluated using clinical scores.

RESULTSThe clinical scores were significantly decreased after immunoadsorption therapy, showing a significant difference from that in the hormonal treatment group (P<0.05). The serum TNF-a and IL-18 levels were significantly higher in the two patient groups than in the control group (P<0.05), but in the former two groups, their levels were significantly lower in immunoadsorption therapy group (P<0.01).

CONCLUSIONImmunoadsorption therapy eliminates the inflammatory cytokines and free radicals as well as the circulating autoantibodies to improve the clinical symptoms of MG.

Adult ; Case-Control Studies ; Female ; Hormones ; therapeutic use ; Humans ; Immunosorbent Techniques ; Interleukin-18 ; blood ; Male ; Middle Aged ; Myasthenia Gravis ; blood ; therapy ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo investigate the significance of trophinin expression in human oocytes and preimplantation embryos.

METHODSThe expression of trophinin in 9 human oocytes, 16 blastomeres and 12 blastocysts were detected by indirect immunofluorescence assay and observed under laser scanning confocal microscope.

RESULTSHuman oocytes, blastomeres and blastocysts were all positive for trophinin expression, and the positivity intensified significantly in the course of the embryonic development (P<0.05).

CONCLUSIONTrophinin may play an important role in human embryo implantation by mediating homophilic adhesion between the embryo and the endometrium during the implantation window.

Blastocyst ; metabolism ; Cell Adhesion Molecules ; biosynthesis ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Microscopy, Confocal ; Oocytes ; metabolism ; Pregnancy