OBJECTIVE To explore the potential neurotoxicity of nano-alu mina (<50 n m)in vivo, we treated the ICR mouse with the nano-alu mina to investigate the mitochondrial da mage of nerve cells on morphology and function.METHODS Adult male mice were exposed to nano-alu mina (<50 n m)of 0,25,50 and 75 mg·kg -1 by nasal instillation for 1 month.Then we observed the mitochondrial ultra-structure of the nerve cells in CA3 region of hippoca mpus,and measured the mean dia meter in every group.The activities of Na +-K +-ATPase and Ca2 +-Mg2 +-ATPase were tested by the determination of the inorganic phosphorus,which was the deco mposition product of ATPase.Western blot analysis was used to detect the expression of COX-Ⅳ,Beclin1 ,LC3Ιand LC3Ⅱ.RESULTS Co mpared with 0 and 25 mg·kg -1 groups exposed to Al2 O3 nanopartilces (Al2 O3 NPs),the mitochondria of CA3 region in hip-poca mpus in 50 mg·kg -1 group beca me ede matous and swollen with sparse and broken cristae sur-rounding the nuclear,and the mean dia meter was higher(0.49 ±0.02 μm,P <0.05).But co mpared with 50 mg·kg -1 group,the mitochondria in 75 mg·kg -1 group beca me s maller with inner cristae of high density,and the mean dia meter was lower(0.36 ±0.02 μm,P<0.05).The enzy me activity of the mito-chondria in cerebral cortex decreased dose-dependently with exposure,the activities of Na +-K +-ATPase in 50 and 75 mg·kg -1 groups(6.37 ±0.22 kU·g -1 protein,5.48 ±1 .53 kU·g -1 protein)and Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups (3.21 ±0.99 kU·g -1 protein,3.28 ±0.15 kU·g -1 protein)were lower than the 0 mg·kg -1 group(P<0.05).Meanwhile,the Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups showed lower activities in co mparison with the 25 mg·kg -1 group.The 75 mg·kg -1 group expressed higher level of the COX-Ⅳ protein 1 .35 ±0.66(P<0.05)than other groups.Both expression of Beclin1 protein and rate of LC3Ⅱ/LC3Ⅰin 75 mg·kg -1 group were more than the 0 mg·kg -1 group. CONCLUSION The mitochondrial dysfunction may be the potential neurotoxicity of nano-alu mina,and the da maged mitochondria were cleared by autophagy.

Objective To explore the correlation between serum hepatitis B virus (HBV) X antigen/antibody (HBxAg-wild/HBxAb-wild,and HBxAg-mutant/HBxAb-mutant) and the disease progression in patients with chronic HBV infection.Methods A direct enzyme immunosorbent asssay (ELISA) was performed to detect HBxAb using recombinant antigen,and a double antibody sandwich ELISA assay to detect HBxAg using monoclonal antibody and specific rabbit polyclonal antibody.HBxAg-wild/HBxAb-wild and HBxAg-mutant/HBxAb-mutant were tested in sera from cases at different stages of chronic HBV infection.A chi-square test was employed to examine statistical significance.Results The positive rates of HBxAg-wild and HBxAg-mutant in the chronic asymptomatic HBV carriers,chronic hepatitis,hepatitis B-related cirrhosis and liver cancer were 6.2％ (2/32),10.7％ (3/28),28.6％ (6/21),43.6％ (17/39) and 3.1％ (1/32),10.7％ (3/28),33.3％ (7/21),48.7％ (19/39),respectively.The positive rates of HBxAb-wild and HBxAb-mutant in the above mentioned groups were 6.2％ (2/32),21.4％ (6/28),38.1％ (8/21),53.8％ (21/39)and 6.2％ (2/32),25.0％ (7/28),42.9％ (9/21),61.5％ (24/39) respectively.The positive rates of HBxAg-wild and HBxAg-mutant were not significantly different among the above groups (x2 =0.871,0.780,0.565 and 0.317,respectively; all P＞0.05) ; The positive rates of HBxAb-wild and HBxAb-mutant were also similar among all the groups (x2 =0.780,0.709,0.580 and 0.210,respectively; all P＞0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in patients with low viral loads (HBV DNA＜1 × 104 copy/mL) were 36.5％ (23/63),44.4％ (28/63),42.9％ (27/63) and 54.0％ (34/63),respectively,those in patients with high viral loads (HBVDNA≥1×104 copy/mL) were 8.8％ (5/57),15.8％ (9/57),5.3％ (3/57) and 14.0％ (8/57),respectively.The positive rates of HBxAg and HBxAb were significantly higher in cases with low viral loads than those with high viral loads (x2 =12.869,11.522,22.556 and 20.976,respectively; all P＜0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in the HBeAg positive group were 21.7％ (18/83),30.1％ (25/83),22.9％ (19/83) and 32.5％ (27/83),respectively,while those in the HBeAg negative group were 27.0％ (10/37),32.4％ (12/37),29.7％ (11/37) and 40.5％ (15/37),respectively.No significant difference of HBxAg/HBxAb positive rates between HBeAg positive group and HBeAg negative group was noticed (x2 =0.408,0.064,0.638 and 0.722,respectively; all P＞0.05).Conclusions The antigenicity and specificity of HBV X protein remains similar after the occurrence of A1762T/G1764A double mutant in X gene.It is also found that the positive rates of HBxAg and HBxAb increase with disease progression.HBxAg/HBxAb might be promoting factors for tumorigenesis in chronic HBV infection.HBxAg and HBxAb might have negative influence on HBV replication.

ABSTRACT:OBJECTIVE To investigate the role of connexin proteins (Cx), which form gap junctions (GJ), in progression and chemotherapeutic sensitivity of cervical cancer (CaCx). METHODS We analyze the expression of Cx26, Cx30, Cx32 and Cx43 in human specimens consisting of: Normal cervix (n=78), CaCx FIGO stage Ⅰ (n=148), CaCx FIGO stage Ⅱ (n=165). InCaCx cell lines, Hela- Cx32 (induced expression by doxycycline), C- 33A (endogenously express Cx32) and siHa (transiently transfected plasmid with Cx32), we detected the role of Cx32 against tostreptonigrin/cisplatin-induced apopotosisin presence or absence of functional GJ through using GJ inhibitors or low density cultural.Furtherly, we observed the relativity of Cx32 and EGFR expression in human specimens. Also, we detected the role of EGFR signaling pathway in the process of Cx32 anti-apoptosis through suppressed EGFR expression by inhibitors or siRNA sequences in cell lines. RESULTS We firstly demonstrated the expression of Cx32 was highly upregulated and accumulated in cytoplasm in the CaCx specimens, and the degree of upregulation correlated with advanced FIGO stages. Thus,in three human cervical cell lines, Cx32 was shown to suppress apoptosis when GJ formation is inhibited. No matter in cases of CaCx or cell lines, Cx32 expression was highly correlated with expression of EGFR and the EGFR pathway is an essential component of the Cx32-induced anti-apoptotic effect. CONCLUSION Cx32, traditionally tumor suppressive protein, was shown to be tumor protective against chemotherapy through EGFR pathway in a GJ-independent way.

OBJECTIVETo elevate the prognostic value of minimal residual disease (MRD) detection by four-color flow cytometry with the antibody panel in childhood B-cell acute lymphoblastic leukemia (B-ALL).

METHODSThe clinical data of 183 children with newly-diagnosed acute B-ALL and who accepted MRD detection between October 2010 and March 2012 was retrospectively reviewed. According to the detection time and result of MRD, the 183 children were classified into four groups: MRD negative (n=37) and positive (n=18) in the induction chemotherapy and MRD negative (n=113) and positive (n=15) in the maintenance chemotherapy.

RESULTSDuring both induction and maintenance chemotherapy, the percentage of patients at high and median risk in the MRD positive group was higher than in the MRD positive group (P<0.05). In the maintenance chemotherapy group, the 3- year cumulative incidence of relapse in MRD positive patients was higher than negative patients (P=0.04). The Cox's proportional hazards regression analysis showed that insensitive reaction for prednisone (RR=1.005, 95%CI: 0.864-1.170, P=0.032), bone marrow morphology that did not meet M1 on the 15th day (RR=6.454, 95%CI: 2.191-19.01, P=0.002) and MRD&ge;0.01% (RR=1.923, 95%CI: 0.750-4.933, P=0.043) were risk factors for relapse in children with B-ALL.

CONCLUSIONSThe four-color flow cytometry with the antibody panel can distinguish from MRD positive patients from negative patients with B-ALL. The result of MRD detection, as prednisone sensitivity and bone marrow morphology on the 15th day, is also a independent prognostic factor in children with B-ALL.

Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Neoplasm, Residual ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology

At present time, the widely used hepatitis B virus( HBV) vaccines consist of only the small hepatitis B surface antigen expressed in yeast or CHO cells. The frequency of non-responders to these vaccines has increased the demand for a more immunogenic vaccine. Some studies have suggested that the addition of preS region to the vaccine will improve its efficacy. However, the large protein (L) containing the whole preS region can not be effectively expressed in vitro. To overcome this problem, two chimeric contructs, SS1, surface gene containing preS1 region at C-terminus and SS2, surface gene containing preS2 region at C-terminus, were constructed and effectively expressed in our previous studies. Here we further constructed an expression vector containing both SS1 and SS2 expression cassettes by separation and ligation the SS2 cassette to a linearized SS1 expression vector pAO815-SS1. The recombinant vector was transformed into Pichia pastoris GS115 by electroporation. A high-level expression strain (GS115-SS1S2) was established by primary screening for His+ transformants and further analysis for induction products. ELISA results demonstrated that the expressed protein had S, preS1 and preS2 antigenicities simultaneously. Western blotting showed that the product can bind to all of the three antibodies, anti-S, anti-preS1 and anti-preS2. The expression protein was present in the form of particles of 20-35 nm diameter and the yield of recombinant particles reached 300-600 mg/L by fermentation. The SS1 and SS2 polypeptides kept intact in purified particles, suggesting that the stability of preS region has been significantly improved.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Epitopes ; genetics ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Peptide Fragments ; genetics ; immunology ; Pichia ; genetics ; Protein Precursors ; genetics ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo assess the utility of P504S immunohistochemistry in the diagnosis and differential diagnosis of prostatic adenocarcinoma.

METHODSLight microscopy and immunohistochemistry examinations (EnVision staining) were performed in 117 cases of prostatic adenocarcinoma, PIN, AAH, ASAP, BPH and normal prostatic tissue to correlate the morphology and protein expression of P504S, 34betaE12, and P63.

RESULTSSeventy-one of the 78 (91%) cases of prostatic adenocarcinoma stained positive for P504S, with strong cytoplasmic granular staining in most cases, and a weak or intense granular staining along the circumferential luminal and apical cell border membrane in a few cases. Negative P504S immunostaining was observed in 7 of 78 (9%) cases of prostatic adenocarcinoma, all of which were clear cell type prostatic adenocarcinoma. Cases of PIN (9 cases), AAH (6 cases) and ASAP (2 cases) showed various expression levels of P504S. Sixty-five of 68 (96%) cases of normal prostates and BPH were negative for P504S and basal cell hyperplasia cases were also negative.

CONCLUSIONSP504S is a useful marker for microscopic diagnosis of prostatic adenocarcinoma, and immunohistochemistry study using a combination of P504S and 34betaE12/p63 may be of greater benefit in aiding the differential diagnoses.

Adenocarcinoma ; diagnosis ; DNA-Binding Proteins ; Diagnosis, Differential ; Genes, Tumor Suppressor ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Male ; Phosphoproteins ; analysis ; Precancerous Conditions ; diagnosis ; Prostate ; enzymology ; Prostatic Hyperplasia ; diagnosis ; Prostatic Intraepithelial Neoplasia ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Racemases and Epimerases ; analysis ; Trans-Activators ; analysis ; Transcription Factors ; Tumor Suppressor Proteins

Objective To compare the effects of three different methods for extracting short RNA-DNA hybrids, including the TRI reagent method , the phenol saturated with water method and the phenol saturated with Tris buffer method in order to facilitate studies on the biological function of RNA-DNA hybrids .Methods Short RNA fragments modifiedwith FAM at the 5′end and those modified with Cy 5 at the 5′end were synthesized .RNA and DNA fragments were annealed to form RNA-DNA hybrids.They were extracted with the above-mentioned 3 methods respectively .The extracted products were analyzed with electrophoresis .Results and Conclusion Short RNA-DNA hybrids can be extracted by the phenol saturated with water method and by the phenol saturated with Tris buffer method .The results can help study the function of short RNA-DNA hybrids .

OBJECTIVES: The number of metastatic lymph nodes (LNs) and the ratio between the number of metastatic LNs and the total number of retrieved LNs (the LN ratio [LNR]) have been proposed as risk factors for recurrence of papillary thyroid carcinoma (PTC). However, the significance of the number of LNs and the LNR in patients with clinically node negative PTC has not been clearly determined. The purpose of this study is to evaluate their significance. METHODS: We retrospectively analyzed 382 patients with PTC who had undergone total thyroidectomy with prophylactic central neck dissection (CND) between January 2000 and December 2010. We excluded patients with lobectomy, concurrent lateral compartment neck dissection, a follow-up period less than at least 2 years, number of harvested central LNs less than or equal to one, clinically positive LN, distant metastasis, recurrent cancer or other types of malignancy. The correlations between recurrence and various clinicopathologic characteristics including tumor size, extrathyroidal extension (ETE), stage, number of metastatic central LNs, and the LNR were investigated. RESULTS: After a mean follow-up period of 82.2±26.4 months, recurrence occurred in 14 patients (3.7%). Tumor size ≥20 mm, maximal ETE, presence of central LN metastasis, number of metastatic LNs ≥2, and LNR ≥0.31 correlated with recurrence in the univariate analysis. However, tumor size ≥20 mm, maximal ETE, number of metastatic LNs ≥2, and LNR ≥0.31 were significantly associated with recurrence in the multivariate analysis (hazard ratio=6.61, 7.17, 3.43, and 11.23, respectively). CONCLUSION: The LNR and the number of metastatic LNs are independent prognostic risk factors for recurrence in patients with clinically node negative PTC, and these factors can be used to guide postoperative adjuvant therapy and follow-up strategy after prophylactic CND.
Follow-Up Studies ; Humans ; Lymph Nodes* ; Multivariate Analysis ; Neck Dissection ; Neoplasm Metastasis ; Recurrence* ; Retrospective Studies ; Risk Factors* ; Thyroid Gland* ; Thyroid Neoplasms* ; Thyroidectomy

Abstract: Olfactory receptors (ORs) are transmembrane proteins mainly distributed in olfactory sensory neurons of the nasal epithelium, mediating the transmission of real-time sensory signals to the brain to produce smell. Recent studies have reported that ORs can also be expressed in tissues or organs outside the nasal cavity, and are closely related to a variety of biological processes, such as sperm chemotaxis, wound healing, glycolipid metabolism and intestinal secretion. In addition, ORs are closely related to a variety of malignant tumors such as prostate cancer, breast cancer and colorectal cancer, and may affect the occurrence and development of tumors by regulating cell proliferation, apoptosis, migration and invasion. This review provides an overview of the effects of ectopic ORs on the function of various human tissues and organs and assesses their potential value as drug targets for the treatment of human diseases.