Association between index-ring finger length ratio and polymorphisms of 6 phalange-bone development related genes / 解剖学报

Objective To investigate the association of 13 single nucleotide polymorphism(SNP)sites in 6 phalange-bone development related genes[fibroblast growth factor receptor 2(FGFR2),indian hedgehog signaling molecule(IHH),Msh homeobox 1(MSX1),Runx family transcription factor 2(RUNX2),SRY-box transcription factor 9(SOX9),Wnt family member 5A(WNT5A)]with human index-ring finger length ratio(2D∶4D).Methods Digital cameras were used to take frontal photographs of the hands of 731 college students(358 males and 373 females)in Ningxia,and image analysis software was used to mark anatomical points and measure finger lengths of index(2th)and ring(4th);genotyping of 13 SNP sites(rs1047057,rs755793,rs41258305,rs3731881,rs3100776,rs12532,rs3821949,rs45585135,rs3749863,rs1042667,rs12601701,rs1829556,rs3732750)for 6 genes by multiplex PCR;One-Way ANOVA or independent sample t-test indirectly assessed the association between 2D∶4D and 13 SNP sites.Results Both left and right hand 2D∶4D were significantly higher in females than males in Ningxia college students(all P<0.01);no statistically significant differences in genotype and allele frequencies of the 13 SNP sites among different sexes(all P>0.05);among different sexes,male left hand 2D∶4D was significantly associated with the genotype of SOX9 gene rs12601701 site(P<0.05)and right hand 2D∶4D was significantly associated with the genotype of WNT5A gene rs1829556 site(P<0.05);the female right hand 2D∶4D was significantly associated with the MSX1 gene rs12532(P<0.01)and rs3821949(P<0.05)sites genotypes.Conclusion SOX9(rs12601701),WNT5A(rs1829556)and MSX1(rs12532 and rs3821949)gene polymorphisms may be associated with the formation of 2D∶4D in Ningxia population.

Application value of MEX3A,CDX2,MUC2 and MUC5AC in judging cancerous gastric mucosal intestinal metaplasia / 中国现代医生

@#Objective To investigate the correlation between MEX3A and differentiation characteristics of gastric cancer and intestinal metaplasia,and its combination with caudal-related homeobox transcription factor 2(CDX2)and mucin 2(MUC2)and mucin 5AC(MUC5AC)to determine the role of carcinogenic intestinal metaplasia.Methods From January 2010 to December 2014,a total of 410 cases of gastric cancer and paracarcinoma paraffin-embedded tissue samples were selected from the Central Hospital Affiliated to Shenyang Medical College and the Second Hospital Affiliated to Shenyang Medical College.According to pathological diagnosis,they were divided into control group(mild superficial gastritis,79 cases),intestinal metaplasia group(149 cases)and gastric cancer group(182 cases).The expressions of MEX3A,CDX2,MUC2 and MUC5AC were detected by immunohistochemistry.Results MEX3A was highly expressed in gastric cancer group and intestinal metaplasia group,especially diffuse gastric cancer,poorly differentiated gastric cancer and type Ⅲ intestinal metaplasia(P<0.05).CDX2 and MUC2 were highly expressed in gastric cancer group and intestinal metaplasia group,especially intestinal type gastric cancer,highly and moderately differentiated gastric cancer,type Ⅰ and type Ⅱ intestinal metaplasia(P<0.05).The expression of MUC5AC was high in control group and low in gastric cancer group and intestinal metaplasia group,especially in intestinal type gastric cancer,type Ⅰ and type Ⅲ intestinal metaplasia(P<0.05).Gastric cancer and intestinal metaplasia differentiation were negatively correlated with MEX3A and MUC5AC expression,but positively correlated with CDX2 and MUC2 expression(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2,and positively correlated with the expression of MUC5AC in gastric cancer(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2 in intestinal metaplasia(P<0.05),while CDX2 was positively correlated with the expression of MUC2(P<0.05).Conclusion MEX3A is negatively correlated with gastric cancer and intestinal metaplasia differentiation.Gastric cancer is characterized by high MEX3A expression and low CDX2 and MUC2 expression.

Association between index finger and ring finger length ratios and polymorphism of homeobox A11 gene locus among Ningxia college students / 解剖学报

Objective To investigate the association between the index finger and ring finger length ratio (2D ∶ 4D) and of four loci (rs6461992‚ rs6968828‚ rs7801581‚ rs17427875) polymorphism of homeobox (HOX) A11 gene among Ningxia college students. Methods Digit camera was used to collect frontal hand photos of 667 Han college students (348 males and 319 females) from Ningxia province; Image analysis software was used to mark the anatomical points and measure finger lengths of the index and ring fingers of both hands; multiplex PCR was used to detect each locus polymorphisms of HOXA11 gene; statistical software was used to compare and analyze the differences and associations of 2D ∶4D and gene polymorphisms between different genders. Results Among Ningxia Han college students‚ both left hand and right hand 2D ∶ 4D were significantly higher in females than those of in males (all P< 0. 05)‚ and there were no significant sex differences in right-left hand 2D ∶4D; the genotypes and allele frequencies of rs7801581 locus of HOXA11 gene differed significantly between genders (all P < 0. 05)‚ and none of the other locus polymorphisms showed any significant sex differences; only female left hand 2D ∶4D was significantly associated with rs6461992 locus genotype in the relationship between 2D ∶4D and HOXA11 polymorphisms (P<0. 05). Conclusion There were significant sex differences in 2D ∶ 4D among Han college students in Ningxia‚ and the rs6461992 locus polymorphism of HOXA11 gene may be associated with the formation of 2D ∶4D in females.

Dendrobium Officinale Polysaccharide Alleviates Hepatic Fibrosis via Epithelial-Mesenchymal Transition / 中山大学学报(医学科学版)

ObjectiveTo investigate the effect of Dendrobium officinale polysaccharide （DOP）on CCl4-induced hepatic fibrosis（HF）and its mechanism. MethodsA total of 56 male SD rats were randomly divided into seven groups： normal group（NG），model group（MG），colchicine group（CG， 0.1 mg/kg）， Fuzheng Huayu group（FG， 0.45 g/kg），low-dose DOP group（LDG， 0.05 g/kg），middle-dose DOP group（MDG， 0.1 g/kg）and high-dose DOP group（HDG，0.2 g/kg），with 8 rats in each group. HF rat model was established by subcutaneous injection with 40% CCl4 olive oil mixture， every 3-day for 10 weeks. At the end of the sixth week， the drug groups were treated with colchicine， Fuzheng Huayu and DOP solution by gavage respectively， once a day for 4 weeks. NG and MG groups were similarly handled with an equal amount of 0.9 % normal saline. Liver histopathology was detected using hematoxylin-eosin （HE）， Masson and Sirius red staining； blood biochemistry was tested for liver function and four indicators of HF； RT-qPCR and Western Blot were used to measure the expression of α-SMA， Col-I， E-cadherin， and ZEB1 genes and proteins in the liver tissues of rats， respectively. ResultsHE， Masson， and Sirius red staining showed that the liver tissue of MG rats had typical pathologic features of HF， and the degree of HF was alleviated in LDG， MDG， and HDG rats， respectively. Liver function test results showed that the serum AST， TBIL， and AKP levels were significantly lower in LDG， MDG， and HDG， compared with those of the MG （P < 0.05 or < 0.01）. Meanwhile， ALT levels in serum deceased remarkably except in LDG （P < 0.05 or < 0.01）. The four results of HF showed that the serum HA， LN， PC-Ⅲ， and COL-Ⅳ levels in LDG， MDG， and HDG rats were significantly decreased compared with those of the MG （P < 0.05 or < 0.01）. The relative expressions of α-SMA， COL-I， and ZEB1 genes and proteins were significantly decreased in the liver tissues of LDG， MDG， and HDG （P < 0.05 or < 0.01）， and the relative expression of E-cadherin gene and protein increased （P < 0.05 or < 0.01）. In addition， the expressions of HA， α-SMA， COL-I， ZEB1 and E-cadherin were dependent on the dose of DOP. ConclusionDOP alleviated the degree of CCl4 induced HF in rats by inhibiting the epithelial-mesenchymal transition in liver tissue.

Effects of circ-SFMBT2 on Proliferation, Migration and Invasion of Acute Myeloid Leukemia Cells by Regulating miR-491-5p/HOXA9 Axis / 中国实验血液学杂志

OBJECTIVE@#To explore the effects and molecular mechanism of circ-SFMBT2 on the proliferation, migration and invasion of acute myeloid leukemia (AML) cells.@*METHODS@#Bone marrow samples from 35 pediatric AML patients and 35 healthy controls in Henan Provincial Children's Hospital from April 2015 to April 2017 and human bone marrow stromal cell lines (HS-5) and AML cell lines (HL-60, THP-1, U-937 and Kasumi-1) were collected. The expressions of circ-SFMBT2, miR-491-5p and homeobox A9 (HOXA9) in bone marrow samples and cells were detected by RT-qPCR and Western blot. The Pearson method was used to analyze the correlation of circ-SFMBT2, miR-491-5p and HOXA9 mRNA expression levels in bone marrow samples of AML patients. HL-60 cells were cultured in vitro and divided into 5 groups: Control, si-NC, si-circ-SFMBT2, si-circ-SFMBT2+anti-NC and si-circ-SFMBT2+anti-miR-491-5p, HL-60 cells were transfected with si-NC, si-circ-SFMBT2, anti-NC, and miR-491-5p inhibitor with Lipofectamine™ 3000. RT-qPCR and Western blot were performed to detect the expression levels of circ-SFMBT2, miR-491-5p and HOXA9 in cells of each group. The proliferation activity of HL-60 cells in each group was detected by CCK-8 assay at 24, 48 and 72 h after transfection, respectively. The apoptosis rate was detected by flow cytometry. The migration and invasion abilities of cells were detected by Transwell assay. The regulatory roles of circ-SFMBT2, miR-491-5p and HOXA9 in AML cells were verified by dual-luciferase reporter gene assay, RNA pull-down and RNA-binding protein immunoprecipitation (RIP) experiments.@*RESULTS@#The expression levels of circ-SFMBT2 and HOXA9 mRNA were increased in bone marrow samples and cell lines (HL-60, THP-1, U-937 and Kasumi-1) of children with AML (P <0.001), while the expression level of miR-491-5p was significantly decreased (P <0.001). Pearson correlation analysis showed that the expression levels of circ-SFMBT2 and miR-491-5p in bone marrow samples of AML children were negatively correlated (r =-0.905), miR-491-5p was also negatively correlated with HOXA9 mRNA (r =-0.930), while the expression levels of HOXA9 mRNA and circ-SFMBT2 was positively correlated (r =0.911). The overall survival rate of AML children with high expression of circ-SFMBT2 was significantly decreased than those with low expression of circ-SFMBT2 (P <0.05). Silencing of circ-SFMBT2 could greatly up-regulate the expression of miR-491-5p, decrease the expression of HOXA9, inhibit the proliferation, migration and invasion of AML cells, and promote cell apoptosis (P <0.05). Down-regulation of miR-491-5p expression greatly attenuated the inhibitory effects of circ-SFMBT2 silencing on cell proliferation, migration and invasion (P <0.05). Dual-luciferase reporter gene assay, RNA pull-down and RIP experiments confirmed that circ-SFMBT2 could target miR-491-5p and negatively regulate the expression of miR-491-5p in AML, and HOXA9 was the target of miR-491-5p.@*CONCLUSION@#Silencing of circ-SFMBT2 may inhibit the proliferation, migration and invasion of AML cells by regulating the miR-491-5p/HOXA9 axis.

Effects of moxibustion on miRNA-133b, Pitx3/TH, and neurotransmitters in the midbrain of rats with diarrhea-predominant irritable bowel syndrome / 针灸推拿医学（英文版）

Objective: To investigate the mechanism of moxibustion in the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D), by observing the effects of moxibustion at Tianshu (ST25) and Shangjuxu (ST37) on microRNA-133b (miRNA-133b), pituitary homeobox family factor 3 (Pitx3)/tyrosine hydroxylase (TH), and neurotransmitters in the brain tissue of IBS-D rats. Methods: Healthy Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, and a Western medicine group, with 12 rats in each group. Except for the normal group, the IBS-D rat model was established by mother-offspring separation and acetic acid enema combined with restraint stress stimulation in all the other groups. No intervention was performed in the normal and model groups. Mild moxibustion was applied to both Tianshu (ST25) and Shangjuxu (ST37) in the moxibustion group. Rifaximin was given by gavage in the Western medicine group. The physical status of rats in each group was observed at different periods. After the intervention, hematoxylin- eosin staining was performed to observe the histopathological morphology of rat colon; enzyme-linked immunosorbent assay was used to measure the levels of dopamine (DA), noradrenaline (NE), and 5-hydroxytryptamine (5-HT) in plasma, colon, and midbrain tissue of rats; the relative expression levels of miRNA-133b, Pitx3 mRNA, and TH mRNA in the midbrain tissue were measured by real-time fluorescence quantitative polymerase chain reaction, and the relative expression levels of Pitx3 and TH proteins in the midbrain tissue were measured by Western blotting and immunofluorescence. Results: The body weights of rats among groups and at different time points were statistically different (P<0.01). The body weight of the normal group was higher than that of the other groups over time (P<0.01). After modeling, the minimum volume threshold of abdominal withdrawal reflex (AWR) was significantly lower (P<0.01) and the loose stool rate was significantly higher (P<0.01) in the model, moxibustion, and Western medicine groups compared with the normal group; the miRNA-133b expression in the midbrain tissue was significantly lower (P<0.01), the expression levels of Pitx3 and TH in the midbrain tissue were significantly higher (P<0.01), and the levels of DA, NE, and 5-HT in plasma, colon and midbrain tissue were significantly higher (P<0.01). After the intervention, the minimum volume threshold of AWR was significantly higher (P<0.01), the loose stool rate was significantly lower (P<0.01), the miRNA-133b expression was significantly increased (P<0.01 or P<0.05) and the expression levels of Pitx3 and TH were significantly decreased (P<0.01) in the midbrain tissue, the levels of DA, NE, and 5-HT in plasma, colon, and midbrain tissue were significantly reduced (P<0.01) in the moxibustion and Western medicine groups compared with the model group; the levels of 5-HT in the colon and midbrain tissue of the moxibustion group were significantly lower than those in the Western medicine group (P<0.05), and there was no statistical difference compared with the remaining groups (P>0.05). Linear correlation analysis showed that miRNA-133b was negatively correlated with Pitx3 (r<0, P<0.01); Pitx3 with TH, TH with DA, and NE with 5-HT were positively correlated (r>0, P<0.01).Conclusion: Moxibustion at Tianshu (ST25) and Shangjuxu (ST37) improves diarrhea symptoms and visceral hypersensitivity in IBS-D rats. The mechanism may be related to up-regulating miRNA-133b, inhibiting Pitx3/TH, and reducing neurotransmitter expression levels in the midbrain tissue.

Overexpression and clinical significance of PBX3 in acute myeloid leukemia / 安徽医科大学学报

Objective @#To investigate the expression levels and potential clinical significance of pre-B-cell leukemia homologous box 3 (PBX3) in bone marrow samples from acute myeloid leukemia (AML) .@*Methods @#The mRNA expression levels of PBX3 were determined by bioinformatics which analyzed the RNAseq data of 33 malignancies from the cancer genome atlasdatabase． (TCGA) The correlations among the expression levels of PBX3，the clinical parameters and prognosis of AML patients were further analyzed．The differentially expressed genes in the whole transcriptome were analyzed to identify the molecular network in AML caused by PBX3 expression abnormalities. @*Results @#The mRNA expression levels of PBX3 were up-regulated in 12 malignancies，and the altitudes increased most significantly in AML than any other cancer types．Patients with high PBX3 expression showed shorter overall survival and disease-free survival than patients with low PBX3 expression．High PBX3 expression was significantly associated with FLT3，NPM1，and DNMT3A mutation．PBX3 expression was positively correlated with multiple ho- meobox genes (including most HOXA and HOXB genes ，MEIS1 ) ，and the expression levels of these homeobox genes were all negatively correlated with AML patients overall survival．@*Conclusion @#PBX3 high expression in the bone marrow of AML patients is a potential biomarker for poor prognosis ，and it may have extensive interactions with other homeobox genes.

Inhibitory effect of siRNA-Pax6 on biological behavior and epithelial-mesenchymal transition of human lens epithelial cells / 中华实验眼科杂志

Objective:To explore the effect of knockdown of the homeobox gene paired-box 6 ( Pax6) on the biological behavior and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs). Methods:The SRA01/04 human LECs were divided into small interfering RNA-Pax6 (siRNA-Pax6) group transfected with siRNA-Pax6 and siRNA negative control (siRNA-NC) group transfected with disordered siRNA.Cell survival rate was detected by cell counting kit-8 method at 24, 48 and 72 hours after transfection.Cell cycle distribution and apoptosis were analyzed by flow cytometry at 48 hours after transfection.Migratory capability of cells was examined by cell scratch test at 24 hours after transfection.The mRNA relative expression levels of Pax6, α-crystallin A (CRYAA), α-crystallin B (CRYAB), Sox2, α-smooth muscle actin (α-SMA) and E-cadherin were detected by quantitative real-time PCR at 48 hours after transfection.The relative expression of Pax6 protein was detected by Western blot at 48 hours after transfection.Results:There was a significant difference in cell survival rates at different time points between the two groups ( Fgroup=4.776, P<0.05; Ftime=13.535, P<0.05). The cell survival rate of siRNA-Pax6 group was obviously lower than that of siRNA-NC group at 48 and 72 hours after transfection, and the differences were statistically significant (both at P<0.05). Compared with siRNA-NC group, the proportion of cells in G 0/G 1 phase was significantly increased and the proportion of cells in S phase was significantly reduced in siRNA-Pax6 group ( t=9.971, -5.063; both at P<0.05). The cell migration rate of siRNA-Pax6 group was (19.73±6.07)%, which was lower than (70.56±2.97)% of siRNA-NC group, showing a statistically significant difference ( t=-7.245, P<0.05). The relative expressions of Sox2 mRNA and α-SMA mRNA were lower, and the relative expression of E-cadherin mRNA was higher in siRNA-Pax6 group than siRNA-NC group, with statistically significant differences between them ( t=-23.254, -5.294, 6.062; all at P<0.01). The relative expression of CRYAA mRNA and CRYAB mRNA was significantly higher in siRNA-Pax6 group than siRNA-NC group, and the differences were statistically significant ( t=5.521, 8.270; both at P<0.01). The relative expressions of Pax6 mRNA and protein in siRNA-Pax6 group were 0.27±0.01 and 0.24±0.05, respectively, which were both lower than 1.00±0.05 and 1.14±0.10 in siRNA-NC group, showing statistically significant differences ( t=-14.456, -4.458; both at P<0.001). Conclusions:Silence of Pax6 can suppress the proliferation and EMT of human LECs and enhance the expression of crystallin.

Six1 gene methylation status in gastric cancer and relationship with clinicopathological characteristics and prognosis / 中国医师进修杂志

Objective:To detect the methylation status of sine oculis homeobox homolog1 (Six1) in patients with gastric cancer and analyze its relationship with the clinicopathological characteristics and prognosis of patients.Methods:The tumor and para-cancerous tissues of 148 patients with gastric cancer diagnosed and treated in Aerospace Center Hospital from September 2015 to December 2017 were collected. The methylation-specific PCR method (MSP) was used to detect the methylation status of the Six1 gene, and 100 normal people who underwent gastroscopy biopsy during the same period served as the control group. Univariate analysis and multivariate Logistic regression model were used to analyze the relationship between Six1 methylation status and clinical pathological characteristics of patients. Kaplan-Meier survival curve was used to analyze the relationship between Six1 methylation status and prognostic survival in patients with gastric cancer.Results:Six1 gene methylation rate in tumor tissue was lower than that in adjacent tissues or in control group, and the differences were statistically significant: 24.32%(36/148) vs. 89.19%(132/148), 96.00%(96/100)( P<0.05). Univariate analysis showed that Six1 gene methylation rate was higher in patients with tumor diameter <5 cm ( χ2 = 8.79, P = 0.003), TNM stage Ⅰ-Ⅱ ( χ2 = 4.93, P = 0.026), highly differentiated tumor ( χ2 = 8.74, P = 0.013), no lymph node metastasis ( χ2 = 4.64, P = 0.031), no distant metastasis ( χ2 = 4.38, P = 0.036), and no invasion of the serosa ( χ2 = 9.85, P = 0.002), and the differences were statistically significant. Multivariate analysis showed that TNM staging ( OR = 4.397, 95% CI 3.141 - 5.157, P = 0.014), tumor differentiation ( OR = 4.491, 95% CI 3.527 - 6.118, P = 0.007), lymph node metastasis ( OR = 4.208, 95% CI 3.823 - 5.195, P = 0.031), distant metastasis ( OR = 4.225, 95% CI 3.956 - 5.437, P = 0.026), and depth of invasion ( OR = 4.509, 95% CI 3.206 - 5.275, P = 0.011) of patients with gastric cancer were independent risk factors for Six1 gene methylation status. Till to March 2020, the mortality rate of the Six1 gene methylation group was lower than that of the Six1 gene unmethylated group: 44.44%(16/36) vs.71.43% (80/112), the difference was statistically significant ( χ2 = 8.70, P<0.05). The median survival time of gastric cancer patients with Six1 gene methylation was higher than that of Six1 gene unmethylated (49 months vs. 37 months), and the difference was statistically significant ( P = 0.019). Conclusions:There is unmethylation of Six1 gene in patients with gastric cancer, which may be involved with the occurrence of gastric cancer. Patients′ TNM stage, tumor differentiation degree, and lymph node metastasis are independent risk factors for Six1 gene methylation status in gastric cancer patients. The prognosis of gastric cancer patients with Six1 gene methylation is better.

Effects of dabrafenib in treatment of skin melanoma and its influence on the expression of pituitary homeobox 1 protein / 肿瘤研究与临床

Objective:To investigate the therapeutic effect of dabrafenib in treatment of cutaneous melanoma (CMM) and its influence on the expression of pituitary homeobox 1 (PITX1) protein.Methods:A total of 86 patients with CMM admitted to Wuhan First Hospital from November 2016 to December 2017 were selected as the research objects, and all patients were divided into the observation group and the control group according to random number table, 43 cases in each group. The control group received chemotherapy regimen, and the observation group was added with dabrafenib on the basis of the control group. Immunohistochemistry method was used to detect the expressions of PITX1 protein in both groups. The treatment effect, recurrence rate, survival, PITX1 expression positive rate and adverse reactions were compared between the two groups.Results:The treatment effective rate in the observation group was higher than that in the control group [46.51% (20/43) vs. 25.58% (11/43)], and the difference was statistically significant ( χ2 = 3.23, P = 0.043); the disease control rate in the observation group was higher than that in the control group [93.02% (40/43) vs.72.09% (31/43)], and the difference was statistically significant ( χ2 = 5.17, P = 0.023). Compared with the control group, the observation group had better overall survival and progression-free survival, lower recurrence rate, and higher PITX1 positive rate (all P < 0.05); adverse reaction rate in the observation group was lower than that in the control group [11.63% (5/43) vs. 32.56% (14/43)], and the difference was statistically significant ( χ2 = 5.47, P < 0.05). Conclusions:Dabrafenib has a better effect in the treatment of CMM, which can increase the expression level of PITX1, reduce the recurrence rate and the incidence of adverse reactions, and prolong the survival time of patients.

The effect of HOXC10 gene on biological behaviors of glioma cells and mechanism in tumor microenvironment / 中华肿瘤杂志

Objective: To study the effects of Homeobox C10 (HOXC10) on biological characteristics such as migration, invasion and proliferation of glioma cancer cells and to explore the role of HOXC10 gene in glioma microenvironment. Methods: The expression level of HOXC10 in high grade glioma (glioblastoma) and low grade glioma and its effect on patient survival were analyzed by using The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) database. Hoxc10-siRNA-1, HOXC10-siRNA-2 and siRNA negative control (NC) were transfected into U251 cells according to the operation instructions of HOXC10-siRNA transfection. 100 ng/ mL recombinant protein chemokine ligand 2 (reCCL2) was added into the transfection group, and was labeled as HOXC10-siRNA-1+ reCCL2 and HOXC10-siRNA-2+ reCCL2 groups. The expressions of HOXC10 mRNA and target protein in each group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and western blot. The proliferation ability of cells in each group was detected by cell counting kit 8 (CCK8) method. The migration ability of cells was detected by Transwell assay and Nick assay, and cell apoptosis was detected by flow cytometry. The expression of chemokines in each group was detected by multiple factors. Co-incubation assays were performed to determine the role of HOXC10 and chemokine ligand 2 (CCL2) in recruiting and polarizing tumor-associated macrophages (M2-type macrophages). Results: The median expression level of HOXC10 in high grade gliomas was 8.51, higher than 1.00 in low grade gliomas (P<0.001) in TCGA database. The median expression level of HOXC10 in high grade gliomas was 0.83, higher than 0.00 in low grade gliomas (P=0.002) in CGGA database. The 5-year survival rate of patients with high HOXC10 expression in TCGA database was 28.2%, lower than 78.7% of those with low HOXC10 expression (P<0.001), and the 5-year survival rate of patients with high HOXC10 expression in CGGA database was 20.3%, lower than 58.0% of those with low HOXC10 expression (P<0.001). The numbers of cell migration in HOXC10-siRNA-1 group and HOXC10-siRNA-2 group were (45±3) and (69±4) respectively, lower than (159±3) in NC group (P<0.05). The cell mobility of HOXC10-siRNA-1 group and HOXC10-siRNA-2 group at 48 hours were (15±2)% and (28±4)% respectively, lower than (80±5)% of NC group (P<0.05). The expressions of vimentin in HOXC10-siRNA-1 group and HOXC10-siRNA-2 group were (141 740.00±34 024.56) and (94 655.00±5 687.97), N-cadherin were (76 810.00±14.14) and (94 254.00±701.45), β-catenin were (75 786.50±789.84) and (107 296.50±9 614.53), lower than (233 768.50±34 114.37), (237 154.50±24 715.50) and (192 449.50±24 178.10) of NC group (P<0.05). The A value of HOXC10-siRNA-1 group and HOXC10-siRNA-2 group were (0.44±0.05) and (0.32±0.02) at 96 hours, lower than 0.92±0.12 of NC group (P<0.05). The apoptosis rates of HOXC10-siRNA-1 group and HOXC10 siRNA-2 group were (10.23±1.24)% and (13.81±2.16)%, higher than (4.60±0.07)% of NC group (P<0.05). The expression levels of CCL2 in U251 cells in HOXC10-siRNA-1 and HOXC10-siRNA-2 groups were (271.63±44.27) and (371.66±50.21), lower than (933.93±29.84) in NC group (P<0.05). The expression levels of CCL5 (234.81±5.95 and 232.62±5.72), CXCL10 (544.13±48.14 and 500.87±15.65) and CXCL11 (215.75±15.30 and 176.18±16.49) in HOXC10-siRNA-1 and HOXC10-siRNA-2 groups were higher than those in NC group (9.98±0.71, 470.54±18.84 and 13.55±0.73, respectively, P<0.05). The recruited numbers of CD14(+) THP1 in HOXC10-siRNA-1 and HOXC10-siRNA-2 groups were (159.33±1.15) and (170.67±1.15), respectively, lower than (360.00±7.81) in NC group (P<0.05), while addition of reCCL2 promoted the recruitment of CD14(+) THP1 cells (287.00±3.61 and 280.67±2.31 in HOXC10-siRNA-1+ reCCL2 group and HOXC10-siRNA-2+ reCCL2 group, respectively, P<0.05). The expressions level of M2-type macrophage-related gene TGF-β in HOXC10-siRNA-1 group and HOXC10-siRNA-2 group were (0.30±0.02) and (0.28±0.02), respectively, lower than (1.06±0.10) in NC group (P<0.05). The expressions level of M1-related gene NOS2 in HOXC10-siRNA-1 and HOXC10-siRNA-2 were (11 413.95±1 911.85) and (5 894.00±945.21), respectively, higher than (13.39±4.32) in NC group (P<0.05). Conclusions: The expression of HOXC10 in glioma is high and positively correlated with the poor prognosis of glioma patients. Knockdown of HOXC10 can inhibit the proliferation, migration and metastasis of human glioma U251 cells. HOXC10 may play an immunosuppressive role in glioma microenvironment by promoting the expression of CCL2 and recruiting and polarizing tumor-associated macrophages (M2 macrophages).

Long non-coding RNA DUXAP9 promotes the proliferation and metastasis of head and neck squamous cell carcinoma / 口腔疾病防治

Objective@#To investigate the role of long non-coding RNA double homeobox A pseudogene 9 (DUXAP9) in head and neck squamous cell carcinoma (HNSCC) and to evaluate the expression level, molecular function and mechanism of DUXAP9 in HNSCC cells.@*Methods@#Differential expression of lncRNAs between normal and tumor tissues in HNSCC tissues were screened using lncRNA microarray, the expression level of DUXAP9 in HNSCC tissues and its relationship with prognosis were analyzed in the TCGA database. The expression levels of DUXAP9 in HNSCC tissues and cell lines were detected using qRT-PCR. The function in HNSCC cells after DUXAP9 silencing was evaluated using the CCK-8 assay, wound healing assay, Transwell migration assay and subcutaneous xenograft assay in nude mice. Changes in the transcription and translation of epithelial-mesenchymal transition (EMT)-related proteins in head and neck squamous cell carcinoma cells after DUXAP9 silencing were detected using qRT-PCR and Western blot.@*Results@#lncRNA microarray results showed that, compared to adjacent normal tissues, DUXAP9 was abnormally upregulated in HNSCC tissues. Analysis from TCGA database showed that, compared to HNSCC patients with low DUXAP9 expression, HNSCC patients with high DUXAP9 expression had poorer survival. The relative expression of DUXAP9 in HNSCC tissues and 4 HNSCC cell lines increased compared to paired adjacent normal tissues as detected using qRT-PCR. Silencing DUXAP9 significantly inhibited the proliferation, migration and expression of EMT-related genes in HNSCC cells. The silencing of DUXAP9 significantly inhibited subcutaneous tumorigenesis of the HNSCC cell line CAL27 in nude mice.@* Conclusion@#Silencing DUXAP9 significantly inhibited the proliferation of HNSCC cells and subcutaneous xenografts in nude mice. DUXAP9 may mediate the migration of head and neck squamous cell carcinoma cells via the EMT pathway.

Research status and prospect of basic application of islet organoid / 器官移植

Organoids are tissue structures, generated from pluripotent stem cells and cultured in vitro, which form self-organize and recapitulate tissues with similar structure and function to the original organs. Organoids have similar appearance and function to the original tissues, and have been widely applied in basic research and clinical trial. At present, the organoids of liver, kidney, islet, brain, intestine and other organs have been successfully cultivated. The use of islet organoid is a hotspot in the field of organoid research. However, islet organoid is currently applied in basic research because rejection after organ transplantation and other issues remain unresolved. In this article, the origin, development and basic application of islet organoid were reviewed, aiming to provide reference for the transformation from basic research of islet organoid into clinical application as well as the treatment of diabetes mellitus.

Molecular mechanism of antiglioma effect of curcumin / 中华神经医学杂志

Objective:To investigate the molecular mechanism of antiglioma effect of curcumin.Methods:Cell experiment: (1) U251MG and SHG-44 cells at logarithmic growth phase were treated with 10 μmol/L curcumin (curcumin group) or same volume of dimethyl sulfoxide solution (control group); cells were transfected with negative control small interfering RNA (siRNA) and long non-coding RNA (lncRNA) H19 siRNA (negative control siRNA group and H19 siRNA group); cells were transfected with negative control siRNA and H19 siRNA, respectively, and then, they were treated with 10 μmol/L curcumin (negative control siRNA+curcumin group and H19 siRNA+curcumin group); the H19 siRNA was co-transfected with negative control miR inhibitor or miR-491-5p inhibitor into these cells (H19 siRNA+negative control inhibitor group and H19 siRNA+miR-4915p inhibitor group); H19 siRNA+negative control miR inhibitor or H19 siRNA+miR-491-5p inhibitor were co-transfected into the cells, and then, they were treated with 10 μmol/L curcumin (H19 siRNA+negative control inhibitor+curcumin group and H19 siRNA+miR-491-5p inhibitor+curcumin group); the cells were co-transfected with miR-491-5p mimic+blank plasmid or miR-491-5p mimic+HOXA9 overexpression plasmid, and then they were treated with 10 μmol/L curcumin (miR-491-5p mimic+blank plasmid+curcumin group and miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group); real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of H19, miR-491-5p, and HOXA9; CCK-8 assay was used to detect the cell proliferation; flow cytometry was used to detect the cell apoptosis; plate cloning method was employed to detect the number of cell clone formation; Transwell assay was used to detect the cell migration; and the HOXA9 protein expression was measured by Western blotting. (2) The 293T cells at the logarithmic growth phase were chosen; the negative control miRNA mimics or miR-491-5p mimics combined with wild-type H19, wild-type HOXA9 3'-UTR plasmid vectors were co-transfected into the cells, respectively (negative control mimic+wild type H19 group and miR-491-5p mimic+wild type H19 group, negative control mimic+wild type HOXA9 3'-UTR group and miR-491-5p mimic+wild type HOXA9 3'-UTR group); the luciferase activity was detected by dual luciferase reporter experiment. (3) Thirty specimens from glioma patients (glioma group) underwent surgical resection and pathologically confirmed in our hospital from May 2017 to May 2019 and 30 normal brain tissue specimens obtained during decompression (normal group) at the same period were chosen; the mRNA expressions of H19, miR-491-5p, and HOXA9 were detected by qRT-PCR, and the HOXA9 protein expression level in these specimens was detected by Western blotting. (4) Twenty-four nude mice were randomly divided into negative control short hairpin RNA (shRNA) group, H19 shRNA group, negative control shRNA+curcumin group, and H19 shRNA+curcumin group ( n=6); U251MG cells stably transfected with negative control shRNA or H19 shRNA were intraperitoneally injected, respectively, into the mice; and 60 mg/kg curcumin was injected on the next d; the tumor volume was measured on the 7 th, 11 th, 15 th, 19 th, 23 rd, and 27 th d of rearing; and the H19, miR-491-5p and HOXA9 mRNA expressions in the tumor tissues were detected by qRT-PCR; the HOXA9 protein expression was detected by Western blotting. Results:(1) When curcumin group comparing with control group, and H19 siRNA group comparing with negative control siRNA group, U251MG and SHG-44 cells had significantly decreased miR-491-5p mRNA and protein expressions, and significantly increased miR-491-5p mRNA expression ( P<0.05); as compared with that in the H19 siRNA+negative control inhibitor group, the HOXA9 mRNA and protein expressions in U251MG and SHG-44 cells of H19 siRNA+miR-491-5p inhibitor group were significantly higher ( P<0.05). When curcumin group comparing with control group, H19 siRNA group comparing with negative control siRNA group, H19 siRNA+curcumin group comparing with negative control siRNA+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly decreased cell proliferation rate, significantly increased apoptosis rate, significantly reduced number of cell clone formation, and significantly reduced cell migration number ( P<0.05). When H19 siRNA+miR-491-5p inhibitor+curcumin group comparing with H19 siRNA+negative control inhibitor+curcumin group, miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group comparing with miR-491-5p mimic+blank plasmid+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly increased cell proliferation rate, significantly reduced apoptosis rate, significantly increased number of cell clone formation, and significantly increased cell migration number ( P<0.05). (2) When miR-491-5p mimic+wild-type H19 group comparing with negative control mimic+wild-type H19 group, miR-491-5p mimic+wild-type HOXA9 3'-UTR group comparing with negative control mimic+wild-type HOXA9 3'-UTR group, the cell luciferase activity was significantly reduced ( P<0.05). (3) As compared with those in the normal group, the H19 and HOXA9 mRNA expressions and HOXA9 protein expression in the glioma group were significantly increased, and the miR-491-5p mRNA expression was significantly reduced ( P<0.05). (4) On the 27 th d of rearing, when H19 shRNA group comparing with negative control shRNA group, and H19 shRNA+curcumin group comparing with negative control shRNA+curcumin group, the tumor volume was significantly reduced, the miR-491-5p mRNA expression in the tumor tissues was significantly increased, and the H19 mRNA, HOXA9 mRNA and protein expressions were significantly reduced ( P<0.05). Conclusion:Curcumin may inhibit the cell proliferation and migration and promote the apoptosis of glioma cells through lncRNA H19/miR-491-5p/HOXA9 axis.

Knockdown of Homeobox D12 Inhibits 7 Factor-Induced Somatic Cell Reprogramming / 中国生物化学与分子生物学报

Differentiated cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by overexpressing defined transcription factors. The process of reprogramming requires the interaction of various transcription factors to regulate the transformation of cell fate. Hoxd12 (Homeobox D12) is one of the transcription factors regulating the embryonic development of vertebrates, and it plays an outstanding role in the development of the limb, body axis formation, and cell signal transduction. However, any roles of Hoxd12 may play in the somatic cell reprogramming and the pluripotency of embryonic stem cells (ESCs) have not been reported. In this study, we firstly used 7 factors (Sall4-Esrrb-Jdp2-Glis1-Mkk6-Nanog-Kdm2b) and Yamanaka factors (Oct4-Klf4-Sox2) as the research model, combined with RNA interference (shRNA) and gene overexpression, to explore the mechanism of Hoxd12 in somatic cell reprogramming. Moreover, we used CRISPR/Cas9 gene editing to construct Hoxd12 knockout embryonic stem cell lines, and combined embryoid body formation (EB) and RNA sequencing (RNA-seq) to explore the function of Hoxd12 in the pluripotency of ESCs. The conclusions are as follows: (1) Knocking down of Hoxd12 inhibits 7 factor-induced reprogramming (

SIX1 Promotes Proliferation and Invasion of Breast Cancer Cells / 中国生物化学与分子生物学报

Sine oculis homeobox homolog 1 (SIX1) is involved in the regulation of many kinds of tumors and plays an important role in the occurrence and development of breast cancer, but the exact mechanism remains to be explored. In this study, we analyzed the expression of SIX1 in breast cancer, and investigated the role of SIX1 in the proliferation and invasion of breast cancer cells. TCGA database and GEPIA2 showed that the expression of SIX1 in breast cancer was significantly higher than that in normal tissues, which was confirmed in different molecular subtypes of breast cancer, including Basal-like, HER2, Luminal A and Luminal B (P < 0. 05). Besides, the analysis of HPA database also showed that the expression of SIX1 was significantly upregulated in breast cancer, and the difference was statistically significant (P < 0. 001). After interfering with the expression of SIX1 in the breast cancer cell line MDA-MB-231 and overexpressing SIX1 in MCF-7, the growth curve and EdU results showed that the proliferation of breast cancer cells was inhibited after knocking down SIX1, while overexpression of SIX1 could promote the proliferation ability (the growth curve assays: P < 0. 05; EdU assays: P < 0. 001). Besides, Transwell assays showed that SIX1 could enhance the invasion ability of breast cancer cells (P < 0. 001). In TCGA database we defined the high and low expression populations according to the upper and lower quartiles of SIX1 gene expression, and differentially expressed genes were found to be associated with metabolism and stem cell regulatory pathways. For further confirmation, high-throughput RNA deep sequencing (RNA-seq) was conducted using SIX1-knockdown cells, and analysis also showed that SIX1 was closely related to metabolism, stem cell regulation and EMT pathways. We selected several representative genes, MYC, SNAI2 and EGFR, to examine their associations with SIX1 expression and prognosis in patients with clinical breast cancer using published GEO datasets and KM-plotter, we found that SIX1 was positively correlated with the expression of MYC, SNAI2 and EGFR, and the high expression of SIX1, MYC, SNAI2 and EGFR is not conducive to the survival of breast cancer patients. GCBI, GeneMANIA and String online tools were conducted to predict the associated genes, lncRNA and miRNA of SIX1. Collectively, our study initially revealed the role of SIX1 in breast cancer and its regulatory mechanism, providing new insights for further studies.

Long Non-coding RNA AC009686.2 Promotes Breast Cancer Cells Proliferation and Invasion / 中国生物化学与分子生物学报

Long non-coding RNAs (LncRNAs) , as regulators of a class of gene expression, play a key role in the development of various types of tumor.We analyzed the TCGA database and found that the expression of LncRNA AC009686.2 in breast cancer tissues was significantly higher than that in normal tissues, and was positively correlated with the poor prognosis of breast cancer patients.qRT-PCR analysis showed that the expression of LncRNA AC009686.2 in breast cancer cells was significantly up-regulated, and the expression level of LncRNA AC009686.2 in MCF7, T47D, ZR7530, BT549, HCC1937, MDA- MB-231 and SKBR3 eells was 6.58, 5.66, 7.29, 9.06, 6.89, 11.17 and 5.38 folds of that in MCF10 A eells, respectively.LncRNA AC009686.2 knockdown in MDA-MB-231 and BT549 cells which expressed relatively high LncRNA AC009686.2 significantly inhibited cell proliferation, colony formation and invasion, and induced cell G,/S phase arrest.The clone inhibition rates of MDA-MB-231 and BT549 cells with LncRNA AC009686.2 knockdown were 0.496%, 0.438% and 0.495%, 0.353% of the control group, respectively.LncRNA AC009686.2 knockdown also down-regulated protein levels of cyclinD2 and ZEB1.However, overexpression of ZEB1 could significantly reverse the decrease of cell invasion ability caused by LncRNA AC009686.2 knockdown.We further analvsed in the software JASPAR database and found that LncRNA AC009686.2 promoter had ZEB1 binding site, and overexpression of ZEB1 could down-regulate the expression level of LncRNA AC009686.2 in breast cancer cells.In conclusion, LncRNA AC009686.2 which highly expressed in breast cancer, promotes cell proliferation and invasion by up-regulating cyclinD2 and ZEB1 expression, while ZEB1 positively regulates LncRNA AC009686.2 expression.This study will provide a theoretical basis for elucidating the role of LncRNA AC009686.2 in breast cancer and related molecular mechanisms.

SNHG6 promotes invasion and metastasis of esophageal squamous cell carcinoma TE1 cells via upregulating ZEB1 / 中国肿瘤生物治疗杂志

@# Objective: To explore the roles and mechanisms of long non-coding RNA (lncRNA) small nucleolar RNA host gene 6 (SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma (ESCC). Methods: Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines (TE1, Yes-2, Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration andinvasionofTE1cells,respectively.Westernblottingwasusedtodetecttheexpressions of MMP-2, MMP-9andZEB1 protein before and after knockdownofSNHG6inTE1cells.Results:SNHG6washighlyexpressedinESCC tissues, compared to para-carcinoma tissues (P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6siRNA (all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.01). The expressions of ZEB1, MMP-2and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.

The Role of ISL1 and LHX5 LIM Homeobox Genes in Bladder Tumourigenesis

@#Introduction: Lin-11, Isl-1 and Mec-3 domains (LIM) homeobox genes are among the most important sub-families of homeobox genes. These genes are thought to play an important role in cancer. In this study, the protein expression of these genes was examined in urothelial carcinoma of the bladder. The expression pattern of Islet-1 (ISL1) and LIM homeobox 5 (LHX5) across different cancer stages and grades, as well as the association between the protein expression of these genes and patient demographics and clinicopathological features, were examined. Methods: A total of 100 formalin-fixed paraffin-embedded urothelial carcinoma tissues were selected from the Department of Pathology, Hospital Kuala Lumpur and the protein expression of ISL1 and LHX5 was determined using immunohistochemistry. Results: Positive expression of ISL1 and LHX5 was detected in 94% and 98% of the samples, respectively. There were no distinct LHX5 expression patterns associated with different cancer stages, but the proportion of high-expressing tumours was higher in high-grade tumours. In addition, there was a significant association between the expression of LHX5 and tumour grade. The proportion of tumours expressing high levels of ISL1 was found to be highest in later stage tumours. Conclusion: The high percentage of tumours expressing both these genes suggests that ISL1 and LHX5 play an important role in bladder tumourigenesis across multiple stages.

Effect of combined treatment with acupuncture, moxibustion and medication on endometrial receptivity and expression of serum HOXA10 in polycystic ovary syndrome of kidney deficiency and blood stagnation / 中国针灸

OBJECTIVE@#To compare the clinical effect of the combined treatment of acupuncture, moxibustion, Chinese herbal medicine and western medication and simple western medication on polycystic ovary syndrome (PCOS) of kidney deficiency and blood stagnation pattern and explore the effect on endometrial receptivity and the expression of serum homeobox gene A10 (HOXA10).@*METHODS@#A total of 60 patients with PCOS of kidney deficiency and blood stagnation pattern were randomized into a combined treatment group and a western medication group, 30 cases in each one. In the western medication group, on the fifth day of menstruation, clomiphene citrate tablets were taken orally, 50 mg each time, once daily, consecutively for 5 days. On the day when the follicle diameter was ≥ 18 mm, chorionic gonadotrophin for muscular injection, a dose of 10 000 U was given. Before sleep, the aspirin enteric-coated tablets were taken orally, 50 mg (except during menstruation). In the combined treatment group, on the base of the treatment as the western medication group, acupuncture and moxibustion were adopted and the Chinese herbal for tonifying the kidney and activating blood circulation was taken orally. The acupoints were Guanyuan (CV 4), Qihai (CV 6), Zusanli (ST 36), Sanyinjiao (SP 6), Zigong (EX-CA 1), etc. Acupuncture was remained for 30 min each time, once every two days and discontinued during menstruation. Chinese herbal was given from the 3rd day of menstruation till the onset of the next menstruation, one dose each day. After consecutive treatment for 3 menstrual cycles in the two groups, the real-time polymerase chain reaction (RT-PCR) method was adopted to determine the expression of serum HOXA10 before and after treatment in the patients of the two groups. The endometrial thickness at ovulatory phase, uterine arterial flow 7 days after ovulation [including uterine arterial pulsatility index (PI), resistance index (RI), peak systolic velocity (PSV)/end diastolic velocity (EDV), meaning S/D], pregnancy rate and the score of Chinese medicine symptoms before and after treatment were compared in the patients between the two groups.@*RESULTS@#① After treatment, the expression of serum HOXA10 was higher than that before treatment in the patients of the two groups (@*CONCLUSION@#The combined treatment with acupuncture, moxibustion and medication effectively improves endometrial receptivity and uterine arterial flow in the patients with PCOS of kidney deficiency and blood stagnation pattern and increases pregnancy rate. The therapeutic effect is better than the simple western medication and its mechanism is probably related to the regulation of serum HOXA10 expression.