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.
Acupuncture Points ; Acupuncture Therapy ; Female ; Genes, Homeobox ; Homeobox A10 Proteins ; Humans ; Kidney ; Moxibustion ; Polycystic Ovary Syndrome/genetics* ; Pregnancy

In recent years, Research shows that cluster A of Hox gene family is a group of master genes for proliferation and differentiation of hematopoietic stem/progenitor cells (HSPCs), which influence on the number of hematopoietic stem cells and the differentiation of HSPCs into erythroid, myeloid, megakaryocytic and lymphocytic lineages, and are closely related with the pathogenesis of leukemia. In this review, the effects of gene Hox on proliferation and differentiation of HSPCs were concerned, while the epigenetics alterations of cluster A in Hox gene family as well as its coexistence with Non-HOX and other fusion genes were also discussed. This made cluster A in Hox gene family plays a regulatory role in pathogenesis of leukemia.
Cell Differentiation ; genetics ; Genes, Homeobox ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukemia ; genetics

This study was purposed to construct the recombinant retroviral vector containing human homeobox gene hoxA10 and to establish the packaging cell lines which stably produce viruses. The whole coding region of hoxA10 gene was amplified by PCR and inserted into the retroviral vector MSCVneo. The recombinant vector was identified by DNA sequencing. The recombinant and control retroviral vectors were transfected into the packaging cell line PT67 by liposome Lipofectamine(TM) 2000. These cell lines stably producing retrovirus were isolated following G418 selection. The viral suspension was harvested and the viral titer was determined by NIH3T3 cells. The results showed that the recombinant retroviral vector was proved to encode hoxA10 genes by sequencing. The cell lines efficiently producing virus were screened by G418 and designated as PT67/MSCVneo and PT67/MSCVneo-hoxA10. The titers of them were 5 x 10(5) CFU/ml and 4 x 10(4) CFU/ml respectively. It is concluded that the recombinant retroviral vector containing homeobox gene hoxA10 and the stably packaging cell lines which efficiently and correctly produce viruses are successfully constructed, which provides a basis for further exploration of the hoxA10 gene function in the regulation of hematopoiesis.
Cell Line ; Genes, Homeobox ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Retroviridae ; genetics ; Transfection

In recent years, many studies have shown that the HOX gene family involved in the proliferation, differentiation and maturation of hematopoietic progenitor cells. There is a close relationship between the expression of HOX gene family and acute myeloid leukemia (AML). Most of the HOX genes can promote the proliferation and inhibit the differentiation of the hematopoietic progenitor cells, but the mechanism leading to acute myeloid leukemia remains unknown. This review focuses on the relationship between HOX gene and acute myeloid leukemia and the possible mechanisms leading to acute myeloid leukemia.
Animals ; Gene Expression Regulation, Leukemic ; Genes, Homeobox ; Humans ; Leukemia, Myeloid, Acute ; genetics

OBJECTIVE: To investigate the effect and molecular mechanism of miR-142-3p to the proliferation, cycle and apoptosis of acute B lymphocytic leukemia (B-ALL) cells by regulating the homeobox gene 5 (HOXA5) expression. METHODS: Real-time fluorescence quantitative PCR was used to detect the expression levels of miR-142-3p and HOXA5 in human B-ALL cell Nalm6 cell line and human B lymphoblast Hmy2-cir cells. Nalm6 was transfected by using liposome transfection technology, miR-142-3p mimic, pcDNA-HOXA5 overexpression plasmid, miR-142-3p mimic+pcDNA-HOXA5 overexpression plasmid, and control. The binding site of HOXA5 and miR-142-3p was predicted according to microRNA.org, and the targeting relationship between miR-142-3p and HOXA5 gene was detected by double luciferase reporter gene experiment. The effect of miR-142-3p to the proliferation of Nalm6 cells was detected using the Cell Counting Box-8 (CCK-8) method and cell clone formation experiments. Flow cytometry was used to detect the effects of miR-142-3p to cell cycle distribution and apoptosis of Nalm6 cells. The expression levels of cell cycle-related proteins, including G RESULTS: Compared with Hmy2-cir cells, miR-142-3p showed low expression in Nalm6 cells and HOXA5 showed high expression (P<0.05). MiR-142-3p and HOXA5 3'-UTR showed complementary binding regions, the luciferase activity of miR-142-3p mimic and wild-type HOXA5 3'-UTR was significantly lower than that of miR-142-3p negative control and wild-type HOXA5 3'-UTR (P<0.05). The proliferation of Nalm6 cells and the number of cell clones could be inhibited by miR-142-3p mimic after 48 and 72 hours of transfection (P<0.05), which causing G CONCLUSION MiR-142-3p can inhibit the proliferation of Nalm6 cells by targeting down-regulation the expression of HOXA5, arrest the G
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Genes, Homeobox ; Homeodomain Proteins/genetics* ; Humans ; Leukemia, B-Cell/genetics* ; MicroRNAs/genetics*

OBJECTIVETo clone core promoter regions of iroquois homeobox gene 1 (IRX1) gene and evaluate the regulatory mechanism of IRX1 transcription.

METHODSUpstream sequence from transcriptional start site was predicted using bioinformatics methods. Serial deleted fragments from IRX1 promoter sequences were amplified by PCR and luciferase reporter plasmids were constructed. The luciferase intensity was analyzed after transferring reporters into GES-1 gastric mucosa cell line.

RESULTSThe promoter of IRX1 was predicted to be within 700 bp from the 5'-flanking region of IRX1 gene. Eight serial deleted luciferase reporter plasmids were constructed. The transcriptional activity of different fragments was expressed as following: p-416>p-584>p-715>p-350>p-687>p-320>p-188>p-92. Except p-320 and p-188, the transcriptional activity of other 6 fragments was higher than that of PGL3-basic plasmid. The transcriptional activity was the highest in p-416 and decreased sharply from p-320 to p-188.

CONCLUSIONSThe fragment p-416 shows the strongest promoter activity. The sequence from -320 bp to -188 bp is identified as core promoter region, which is focused as key sequence for further regulatory analysis, since some binding sites for important transcriptional factors such as Sp1 and TFII D are predicted.

Cell Line ; Cloning, Molecular ; Gastric Mucosa ; cytology ; Genes, Homeobox ; Homeodomain Proteins ; genetics ; Humans ; Promoter Regions, Genetic ; Transcription Factors ; genetics

OBJECTIVETo explore the expression of beta protein 1 (BP1) gene in adult acute leukemia (AL) and its relationship with acute leukemia.

METHODSExpression of BP1 gene mRNA was detected in 70 adult AL, 10 normal controls and HEL cell line, by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSNo detectable BP1 gene was found in peripheral blood or bone marrow cells of normal controls and 20 acute myeloid leukemia (AML) in complete remission (CR) stage. BP1 gene was highly expressed in HEL cell line, 57% (20/35) of AML and 80% (8/10) of AML-M(5) cases. BP1 gene could not be detected in adult acute lymphoid leukemia.

CONCLUSIONBP1 gene was highly expressed in AML. It might be used as a molecular marker of AML.

Acute Disease ; Adolescent ; Adult ; Female ; Genes, Homeobox ; Globins ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; Transcription Factors ; genetics

OBJECTIVETo construct the adenoviral vector containing human Bcl-2 gene and to study the expression of the gene in the spiral ganglion cells (SGC) in vitro.

METHODSHuman Bcl-2 cDNA obtained from the plasmid pUC-CAGGS/Bcl-2 was cloned into the plasmid pAdTrack-CMV. Then, pAdTrack/Bcl-2 was cotransferred with adenoviral backbone vector into E. coli strain BJ5183. The recombinant adenoviral plasmid was identified by restriction analysis with Pac I and transfected into HEK293 cells to package and amplify recombinant adenovirus particles which would be identified by Electron microscope. After the adenovirus infected the rat spiral ganglion cells, the expression of Bcl-2 gene was detected by Western Blot and RT-PCR.

RESULTSThe recombinant AdEGFP/Bcl-2 plasmid was correctly constructed and confirmed by restriction endonuclease analysis. The viral particles in HEK293 cells were identified by Electron microscope. RT-PCR and Western blot showed that Bcl-2 gene was exactly transcripted and expressed in transgene SGC.

CONCLUSIONSThe method based on homologous recombination in bacteria is simple and high efficient. The recombinant adenoviral vector containing human Bcl-2 cDNA was constructed and the transgene SGC expressed human Bcl-2 gene in vitro successfully. It provided foundation for the further study of protection for the impaired SGC by hBcl-2 gene.

Adenoviruses, Human ; genetics ; Animals ; Cells, Cultured ; Genes, Homeobox ; Genes, bcl-2 ; Genetic Vectors ; Humans ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; Spiral Ganglion ; metabolism ; Transfection ; Transgenes

This study was purposed to construct lentivirus vector containing human homeobox gene HOXB4 and explore changes of human umbilical cord mesenchymal stem cells (HUCMSC) after infected with HOXB4 mediated by lentivirus. PCR amplification was performed to obtain HOXB4, which was cloned in lenti-shuttle vector. Four-plasmid lentivirus packaging system was used to transfect HEK293T cells. After 48 h, lentivirus Lenti-HOXB4 was harvested and lentivirus titer was determined. Lenti-HOXB4 was used to infect HUCMSC. The infected cells were observed under inverted fluorescence microscope to determine the optimal multiplicity of infection (MOI). Meanwhile, RT-PCR, immune fluorescence staining, CCK-8 and flow cytometry (FCM) were used to determine the expression of HOXB4 and its effect on cell growth. The results indicated that lenti-HOXB4 was successfully obtained by co-transfecting the 293T cells with four plasmids. The determined virus titer was 3×10(8) TU/ml; when MOI was 20. Lenti-HOXB4 had a high transfection rate in HUCMSC, over 80%. In HUCMSC infected with lenti-HOXB4, the expression of target gene could be detected both at mRNA and protein levels. It could promote the proliferation of HUCMSC. FCM results indicated HOXB4 gene did not significantly influence the surface marker of HUCMSC. It is concluded that HOXB4 gene can promote the high proliferation of HUCMSC and does not significantly influence the expression of the surface marker of HUCMSC.
Cell Proliferation ; Cells, Cultured ; Flow Cytometry ; Genes, Homeobox ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; cytology ; Plasmids ; Transcription Factors ; genetics ; Umbilical Cord ; cytology

OBJECTIVETo identify whether HOX genes can be used as markers of esophageal cancer.

METHODSThe expression of 39 HOX genes in esophageal cancer cell lines was examined. Specific primers were designed and RT- PCR was performed for each HOX gene members in above esophageal cancer cell lines, EC109 and CAES.

RESULTSFifteen out of 39 HOX genes were expressed in esophageal cancer cell lines. They were HOXA2, HOXA7, HOXA9, HOXA10, HOXA13, HOXB7, HOXB9, HOXC4, HOXC5, HOXC6, HOXC8, HOXC9, HOXD9, HOXD10, and HOXD13 respectively. Of them. Eleven genes were overlapped with the ones detected in human esophageal squamous cell carcinoma(ESCC) in our former study.

CONCLUSIONThis study reconfirms our former that some HOX genes are deregulated expressed in ESCC, which provides more positive evidence for their roles in ESCC.

Cell Line, Tumor ; Esophageal Neoplasms ; genetics ; Gene Expression Profiling ; Genes, Homeobox ; genetics ; Humans ; Multigene Family ; RNA ; genetics ; Reverse Transcriptase Polymerase Chain Reaction