Objective:To investigate the role and mechanism of miR-143 in the proliferation and migration of gastric cancer (GC) cells. Methods:Western blot was performed to detect the expression level of avian erythroblastosis oncogene B-3 (ERBB3) in GC tissues, paired non-cancerous tissues, and SGC7901 GC cells. RT-qPCR was conducted to determine the mRNA and miR-143 of ERBB3 quantita-tively. Bioinformatics tools were used to predict the target gene of miR-143. Luciferase reporter assay was carried out to confirm the predicted target gene. Transwell and EdU assays were applied to observe the migration and proliferation of SGC7901 GC cells transfect-ed with miR-143 mimics/inhibitor/NC mimics/inhibitor. Results:Compared with the expression levels of ERBB3 and miR-143 in the paired non-cancerous tissues, the expression level of ERBB3 was upregulated and the expression level of miR-143 was downregulated in GC tissues. In the prediction of the potential target gene, miR-143 could bind to a specific sequence of the 3′-untranslated regions (UTR) of the mRNA of ERBB3. This finding was supported by luciferase reporter assay results. In vitro, ERBB3 protein expression and cell migration and proliferation were suppressed significantly in the SGC7901 cells transfected with miR-143 mimics. By contrast, these processes were remarkably enhanced when the cells were transfected with miR-143 inhibitor. Conclusion:miR-143 can suppress the migration and proliferation of GC cells by downregulating the expression of ERBB3.

Objective To study the clinical significance of PI3Kp110αexpression in breast cancer. Methods The expressions of PI3Kp110α, HER2, PTEN, p?Akt and Ki?67 in invasive ductal carcinoma ( IDC ) , adjacent ductal carcinoma in situ ( DCIS ) and normal breast tissue were detected by immunohistochemistry in 102 cases of breast cancer. The expression of PI3Kp110α in IDC was compared with those in DCIS and normal breast tissues. Correlations between expression of PI3Kp110αand expression of HER2, PTEN, p?Akt and Ki?67 index in IDC were analyzed. Correlation between expression of PI3Kp110αand stage of IDC was studied as well. Results In the normal breast tissues, there were 97 cases (95.1%) with low level and 5 cases (4.9%) with high level expression of PI3Kp110α. In the DCIS tissues, there were 67 cases ( 65. 7%) with low level and 35 cases ( 34. 3%) with high level expression of PI3Kp110α. In the IDC tissues, there were 14 cases (13.7%) with low level and 88 cases (86.3%) with high level expression of PI3Kp110α. The difference between expression of PI3Kp110α in normal breast tissue, DCIS and IDC was significant ( P<0. 001 ) . In the IDC tissues, expression of PI3Kp110α was negatively correlated with expression of HER2 ( rs=-0.213,P=0.032) and PTEN ( rs=-0.197,P=0.047) , while was not significantly correlated with expression of p?Akt ( P=0.119) and Ki?67 index ( P=0.636) . In contrast, expressions of HER?2 and p?Akt were positively correlated with Ki?67 index ( P=0. 001, P=0.035), while expression of HER2 was not correlated with p?Akt (P=0.177). Expression of PI3Kp110αwas negatively correlated with T stage and TNM stage (P=0.003, P=0.016), while not correlated with N stage and M stage(both P>0.05). Expression of HER?2 was positively correlated with T stage (P=0.037), while not correlated with N stage, M stage and TNM stage ( P>0. 05 for all ) . Neither Ki?67 index nor expression of PTEN and p?Akt (P=0.194) were correlated with T stage, N stage, M stage and TNM stage. Conclusions Expression of PI3Kp110α plays an important role in oncogenesis and development of breast cancer. Based on the fact that expression of PI3Kp110αis negatively correlated with expression of HER2 and PTEN and with T stage and TNM stage, we may conclude that increased expression of PI3Kp110αis another independent pathway in breast cancer development, and breast cancer patients with high expression of PI3Kp110α may have a better prognosis.

Objective To study the clinical significance of PI3Kp110αexpression in breast cancer. Methods The expressions of PI3Kp110α, HER2, PTEN, p?Akt and Ki?67 in invasive ductal carcinoma ( IDC ) , adjacent ductal carcinoma in situ ( DCIS ) and normal breast tissue were detected by immunohistochemistry in 102 cases of breast cancer. The expression of PI3Kp110α in IDC was compared with those in DCIS and normal breast tissues. Correlations between expression of PI3Kp110αand expression of HER2, PTEN, p?Akt and Ki?67 index in IDC were analyzed. Correlation between expression of PI3Kp110αand stage of IDC was studied as well. Results In the normal breast tissues, there were 97 cases (95.1%) with low level and 5 cases (4.9%) with high level expression of PI3Kp110α. In the DCIS tissues, there were 67 cases ( 65. 7%) with low level and 35 cases ( 34. 3%) with high level expression of PI3Kp110α. In the IDC tissues, there were 14 cases (13.7%) with low level and 88 cases (86.3%) with high level expression of PI3Kp110α. The difference between expression of PI3Kp110α in normal breast tissue, DCIS and IDC was significant ( P<0. 001 ) . In the IDC tissues, expression of PI3Kp110α was negatively correlated with expression of HER2 ( rs=-0.213,P=0.032) and PTEN ( rs=-0.197,P=0.047) , while was not significantly correlated with expression of p?Akt ( P=0.119) and Ki?67 index ( P=0.636) . In contrast, expressions of HER?2 and p?Akt were positively correlated with Ki?67 index ( P=0. 001, P=0.035), while expression of HER2 was not correlated with p?Akt (P=0.177). Expression of PI3Kp110αwas negatively correlated with T stage and TNM stage (P=0.003, P=0.016), while not correlated with N stage and M stage(both P>0.05). Expression of HER?2 was positively correlated with T stage (P=0.037), while not correlated with N stage, M stage and TNM stage ( P>0. 05 for all ) . Neither Ki?67 index nor expression of PTEN and p?Akt (P=0.194) were correlated with T stage, N stage, M stage and TNM stage. Conclusions Expression of PI3Kp110α plays an important role in oncogenesis and development of breast cancer. Based on the fact that expression of PI3Kp110αis negatively correlated with expression of HER2 and PTEN and with T stage and TNM stage, we may conclude that increased expression of PI3Kp110αis another independent pathway in breast cancer development, and breast cancer patients with high expression of PI3Kp110α may have a better prognosis.

OBJECTIVETo investigate HIV-1 co-receptor usage in patients experienced anti-retroviral therapy (ART) in Anhui and Henan province of China.

METHODSA total of 45 HIV-1 infected individuals who have experienced ART and 109 un-experienced ART patients from Anhui and Henan province, which were called as treatment group and treatment-negative group, were selected as study subjects. HIV-1 strains were isolated from peripheral blood mononuclear cells of whole blood from patients. HIV-1 p24 in the culture supernatant was measured using a commercial enzyme-linked immunosorbent assay (ELISA) kit. HIV-1 co-receptor usage was identified using Ghost cell lines expressing CD4 and the chemokine receptor CCR5 or CXCR4.

RESULTSAmong 45 HIV strains from the treatment group, 22 (48.9%) strains used CCR5 as a co-receptor (R5 tropic strain), 21 (46.7%) strains used CXCR4/CCR5 as a co-receptor (X4/R5 duel tropic strain), and 2 (4.4%) used only CXCR4 as a co-receptor (X4 tropic strain). In 109 strains from treatment-negative group, 96 (88.1%) strains used CCR5 as a co-receptor (R5 tropic strain), 13 (11.9%) strains used CCR5/CXCR4 as a co-receptor use (X4/R5 strain). A significant difference was found between two groups in X4 co-receptor usages (χ(2) = 27.30, P < 0.05). Furthermore, after treated with AZT + DDI + NVP, the HIV-1 CXC4/CCR5 utilization was 59.09% (13/22), meanwhile after treated with D4T + DDI + NVP, the HIV-1 CXC4/CCR5 utilization was 43.48% (10/23), which the difference was not statistical significant (χ(2) = 1.10, P = 0.30).

CONCLUSIONHIV-1 CXCR4/CCR5 co-receptor utilization was higher in ART patients than treatment-negative patients.

Acquired Immunodeficiency Syndrome ; drug therapy ; metabolism ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Cells, Cultured ; Female ; HIV-1 ; isolation & purification ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; metabolism ; Receptors, CXCR4 ; metabolism ; Receptors, HIV ; metabolism

Objective To measure the values of quantitative ultrasound(QUS) of different gestational age preterm infants at birth in Guangzhou area,and compare them with the values of Caucasian preterm infants in order to get insight into the bone development status of preterm infants in Guangzhou area and to evaluate the practicability of QUS.Methods The Omnisense quantitative ultrasound produced by Israel Sunlight company was used to measure the bone speed of sound(SOS) of left tibia of preterm infants born between Jun.2010 and Jun.2012 in Maternity and Children Health Hospital of Huadu District in Guangzhou,and the values of SOS of Caucasian preterm infants was compared.Results There were totally 1039 preterm infants born in Guangzhou area involved in this study,and they were divided into group A,B,C,D by gestational ages:≤ 30 weeks,30 + 1 ～ 32 weeks,32 + 1 ～ 34 weeks,34 + 1 ～ 36 + 6 weeks.The values of SOS of each group at birth were(2892.05 ± 139.17) m/s,(2936.84 ± 137.87) m/s,(2966.65 ± 116.60) m/s and (2988.63 ± 120.74) m/s,separately,and with the increase of gestational age,and there was significant difference of SOS between different gestational age groups(F =15.758,P =0.000).But there was no significant difference of SOS between male and female (F =2.665,P =0.103).Compared with Caucasian preterm infants,the SOS value gap (defined as the Z value) of preterm infants of different gestational age in Guangzhou area was no significant difference(P =0.117),and there was no significant difference between male and female (F =3.494,P =0.062).Conclusions The value of SOS of preterm infants was higher with the increase of maturity of preterm infants.There was no significant difference of SOS between Guangzhou preterm infants and Caucasian preterm infants.And QUS is suitable for clinical evaluation of bone development status of preterm infants.

Objective To knockout the exon51 of DMD gene in HEK293T cells using the CRISPR/Cas9 system. Methods Design the target sequences of sgRNA and clone them into plasmid PX459 respectively; transfer these plasmids into HEK293T cell and extract the total genome DNA; test the activity of sgRNAs with surveyor assay, choose the most efficient one in each end;construct plasmid PX459-2sgRNA and transfer it into HEK293T cells;check whether the exon51 has been knocked known with PCR and T vector sequencing. Results 50% of HEK293T cells' DMD gene exon51 were knocked out,showing a high gene editing efficiency. Conclusions We successfully establish a platform to target knockout the exon51 of DMD gene and provide an important experimental basis for the treatment of DMD and other genetic diseases.

Endothelin-3 (ET-3) is aberrantly expressed in both metastatic melanoma tissues and cultured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic melanoma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cysteine (SPARC) levels were down-regulated together with inhibition of its upstream signaling molecule, NF-κB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the malignant behaviors of human melanoma cells partially by decreasing the expression of SPARC and NF-κB.
Cell Line, Tumor ; Endothelin-3 ; genetics ; Gene Silencing ; Humans ; Melanoma ; genetics ; pathology ; Osteonectin ; genetics

Objective: The aging model of guinea pigs induced by D-galactose was set up to investigate the changes of BK_Ca expression and function on cochlear pericytes and their relationship with age-related hearing loss. Methods: Thirty healthy 8-week-old guinea pigs were randomly divided into three groups, with 10 in each group: D-galactose aging model group, subcutaneous injection of D-galactose (500 mg/kg) daily for 6 weeks; saline control group, the same amount of saline was injected into the neck of the aging model group for 6 weeks; the blank control group, no treatment was performed. The threshold of auditory brainstem response (ABR) was detected. The content of BK_Ca in the perivascular cells of the guinea pig cochlear cells was detected by immunofluorescence technique. The changes of peripheral current density and BK_Ca current were detected by patch clamp technique. The data were analyzed by GraphPad Prism software. Results: Compared with the saline group and the control group, the ABR threshold and the amplitude of the wave I were significantly decreased in the aging model group, and the difference was statistically significant (P<0.01). Compared with the control group, the expression of BK_Ca in the vascular pericytes of guinea pigs in the aging model group was significantly reduced (1.00±0.08 vs 0.27±0.03,the difference was statistically significant P<0.01), and the cell current density and BK_Ca net current value were also significantly reduced with statistically significant (P<0.01). Conclusions D-galactose can successfully induce guinea pig aging model, in which BK_Ca expression decreases and net current value decreases in pericytes of cochlear striavascularis, and changes in BK_Ca expression and function may be related to age-related hearing loss.

Myasthenia gravis (MG) is a prototypical antibody-mediated neurological autoimmune disease with the involvement of humoral immune responses in its pathogenesis. T follicular helper (Tfh) cells have been implicated in many autoimmune diseases. However, whether and how Tfh cells are involved in MG remain unclear. Here, we established and studied a widely-used and approved animal model of human MG, the rat model with acetylcholine receptor alpha (AChRα) subunit (R-AChR)-induced experimental autoimmune myasthenia gravis (EAMG). This model presented mild body-weight loss 10 days after the first immunization (representing the early stage of disease) and more obvious clinical manifestations and body-weight loss 7 days after the second immunization (representing the late stage of disease). AChR-specific pre-Tfh cells and mature Tfh cells were detected in these two stages, respectively. In co-cultures of Tfh cells and B cells, the number of IgG2b-secreting B cells and the level of anti-AChR antibodies in the supernatant were higher in the cultures containing EAMG-derived Tfh cells. In immunohistochemistry and immunofluorescence assays, a substantial number of CD4/Bcl-6 T cells and a greater number of larger germinal centers were observed in lymph node tissues resected from EAMG rats. Based on these results, we hypothesize that an AChR-specific Tfh cell-mediated humoral immune response contributes to the development of EAMG.
Animals ; B-Lymphocytes ; immunology ; Disease Models, Animal ; Female ; Immunity, Humoral ; Lymph Nodes ; immunology ; Myasthenia Gravis, Autoimmune, Experimental ; immunology ; Protein Subunits ; immunology ; Proto-Oncogene Proteins c-bcl-6 ; immunology ; Rats, Inbred Lew ; Receptor Cross-Talk ; Receptors, Cholinergic ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology