OBJECTIVE: To observe the effect of "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique on the reproductive function of the mice with poor ovarian response (POR) and explore the molecular mechanism of acupuncture on early embryos after in vitro fertilization-embryo transfer (IVF-ET) in POR mice. METHODS: Of 70 female C57BL/6 mice, 60 mice with regular estrous cycle were screened and 30 of them were randomly divided into a blank group, a model group, and an acupuncture group, 10 mice in each one. Mice in the model and acupuncture groups were administered with tripterygium glycosides suspension (50 mg·kg^-1·d^-1) via gavage for 2 weeks to prepare POR models; while in the blank group, the mice received an equal volume of 0.9% sodium chloride solution via gavage. After successful modeling, mice in the acupuncture group underwent "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique, once daily, for 20 min each time and lasting 2 consecutive weeks. After intervention completion, subsequently, all the three groups underwent ovulation induction, orbital blood collection, and ovary extraction and fresh denuded oocyte collection. Denuded oocytes, after incubated, together with the sperms from 15 male C57BL/6 mice, were transferred into the oviducts of 30 donor pseudopregnant C57BL/6 female mice via IVF-ET; and the embryonic tissue was collected on day 7 of implantation. After successful modeling and intervention completion, the general conditions of mice in each group were observed, and the estrous cycle disorder rate was compared among the groups. After intervention completion, the numbers of oocyte, ovarian wet weight and ovarian index were recorded in each group; the levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in the serum were measured using ELISA; and ovarian morphology and mitochondrial ultrastructure were examined using HE staining and transmission electron microscopy. The transcriptome sequencing technology was employed to identify differentially expressed genes (DEGs) in early embryos of each group, followed by bioinformatics analysis; and the reversed DEGs with significant difference were verified using quantitative real-time PCR (qRT-PCR). RESULTS: After intervention, compared with the blank group, the mice in the model group presented poor spirits and declined water and food intake, reduced activity, the higher rate of estrous cycle disorder (P< 0.01); the decrease of the numbers of oocyte, ovarian wet weight, and ovarian index (P<0.01), reduced AMH and E2 in the serum (P<0.01), elevated FSH and LH in the serum (P<0.01); more atresia follicles in ovarian tissue, disorganized granulosa cells, reduced the numbers of mitochondria, vacuoles appeared and cristae breakage. When compared with the model group, in the acupuncture group, the spirits of mice, food intake and activity were improved, the rate of estrous cycle disorder was reduced (P<0.01); the numbers of oocyte, ovarian wet weight, and ovarian index were higher (P<0.01, P<0.05), the levels of AMH and E2 in the serum increased (P<0.01), the levels of FSH and LH in the serum decreased (P<0.01); the atresia follicles were reduced, the numbers of follicle increased at all levels, the numbers of mitochondria got higher, vacuoles and cristae breakage were declined. In comparison between the blank group and the model group, and between the model group and the acupuncture group, a total of 258 intersecting DEGs were identified, and 194 DEGs of them exhibited a trend of reversion before and after intervention. The top 5 reversed DEGs with significant difference included dexamethasone-induced Ras-related protein 1 (Rasd1), gene regulated by estrogen in breast cancer 1 (Greb1), leucine-rich repeat-containing G protein-coupled receptor 6 (Lgr6), Fraser syndrome 1 (Fras1), and apolipoprotein D (Apod). Compared with the blank group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues decreased in the model group (P<0.01, P<0.05); and when compared with the model group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues increased in the acupuncture group (P<0.05, P<0.01). These findings were consistent with the DEGs screening results. Gene ontology (GO) analysis revealed that the reversed genes of co-expression were primarily participated in the biological processes such as myoblast differentiation, endocardial cushion development, and cardiac morphogenesis. Kyoto encyclopedia of genes and genomes (KEGG) analysis indicated that the reversed genes of co-expression are enriched in the Wnt signaling pathway, and they were associated with various types of cardiac diseases. CONCLUSION The "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique may ameliorate the decline of ovarian response in mice with POR, promote normal follicle development and ovulation, thereby improve embryo quality and reduce the risk of developmental defects and deformity in the organs such as the embryonic heart. The underlying mechanism may be related to the regulation of the expression pattern of embryonic key genes through the Wnt signaling pathway.
Animals ; Female ; Mice ; Mice, Inbred C57BL ; Acupuncture Therapy ; Ovary/physiopathology* ; Humans ; Fertilization in Vitro ; Transcriptome ; Embryo Transfer ; Infertility, Female/physiopathology*

Objective To investigate the effect and mechanism of osteopontin(OPN)in hepatoma cell migration through galectin-3 binding protein(LGALS3BP).Methods Human hepatoma cell lines SMMC-7721,SMMC-P(stably transfected with empty eukaryotic expression vectors),and SMMC-OPN(stably transfected with the OPN gene)were cultured.mRNA expression levels of OPN and LGALS3BP were measured by RT-qPCR.Western blot assays were used to analyze the relative protein expression of OPN and LGALS3BP and PI3K/AKT pathway.Wound healing assays were performed to explore the cell migration ability.After transfection with LGALS3BP-targeting small interfering RNA(si-LGALS3BP)or negative control small RNA(si-NC)into SMMC-OPN cells,cell migration and relative expression of PI3K/AKT pathway-related proteins were assessed.Results Compared with SMMC-7721 and SMMC-P,the migratory ability of SMMC-OPN cells was significantly reinforced,and expression of LGALS3BP was obviously upregulated at both mRNA and protein levels.Moreover,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins was significantly increased.Wound healing assays showed that the si-LGALS3BP obviously suppressed the migratory ability of SMMC-OPN cells.Furthermore,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins in SMMC-OPN cells was significantly decreased after transfection of si-LGALS3BP.Conclusions OPN activates the PI3K/AKT pathway by upregulating LGALS3BP expression to promote hepatoma cell migration.

Objective:To investigate the status of dyadic coping among patients with lung cancer and their spouses, and explore the related factors of dyadic coping.Methods:A convenience sample of 227 patients with lung cancer and their spouses were selected. The measurements of socio-demographic questionnaire, Dyadic Coping inventory (DCI), the Quality of Relationship index (QRI) and Connor-Davidson Resilience Scale (CD-RISC-10) were used to assess the general information, dyadic coping, relationship satisfaction and resilience, respectively.Results:The DCI score of patients were (3.13±0.89) points; the DCI score of spouses were (3.03±0.93) points; the scores of DCI were significant correlated between patients and their spouses ( r values were 0.248-0.632, P<0.01). The relationship satisfaction ( β values were 0.421, 0.474, P<0.01) and resilience ( β values were 0.374, 0.211, P<0.01) were the significant predictor of dyadic coping. Meanwhile, partner effects for patients′ resilience ( β value was 0.193, P<0.05) and spouse′ relationship satisfaction ( β value was 0.237, P<0.01) were also significant. Conclusions:Moderate dyadic coping were occurred among patients with lung cancer and their spouses. Relationship satisfaction and resilience were the mainly predictors for dyadic coping. To improve relationship satisfaction and enhance resilience might be an effective approach to promote positively dyadic coping.

Objective To analyzer the clinical significance of the Xpert MTB/RIF method for rapidly detecting Mycobacterium tuberculosis(MTB) and rifampicin resistance in clinic.Methods A total of 552 sputum samples were collected from the patients in the Huizhou Municipal Institute of Tuberculosis Prevention and Control and tuberculosis departments in 5 counties and districts from January 2015 to December 2015.Smear microscopic examination ,Xpert MTB/RIF nucleic acid detection and improved Roche culture were performed.The samples of culture positive were performed the drug susceptibility test.The sensitivity ,specificity of various methods for detecting MTB were evaluated.The results of conventional susceptibility test and Xpert MTB/RIF for detec-ting rifampicin resistance were compared and analyzed.Results Compared with the Roche culture method ,the sensitivity and speci-ficity of Xpert MTB/RIF for detecting MTB were 95.3% and 93.1%.In the comparison of drug resistance detection results ,the sensitivity and specificity of rifampicin resistance in all positive samples were 82.1% and 97.8%.In the consistency analysis of Xpert MTB/RIF and improved Roche culture for detecting MTB classification ,kappa=0.688 ,indicating that the results of the two methods had good consistency.Conclusion The Xpert MTB/RIF method can be used in MTB early rapid detection and screening of rifampicin resistance detection ,which is conducive to clinical rapid decision.

Objective To study the expression of fragile histidine triad(FHIT) in oral squamous cell carcinoma(OSCC),and discuss its role and significance in OSCC.Methods Immunohistochemical method was used to detect the expressions of FHIT in 48 cases of OSCC and 26 normal oral mucosa.Results The positive expression rate of FHIT in normal oral mucosa was 76.92%(20/26).Among 48 OSCC patients,the positive rate of FHIT(43.75%) was lower than that in normal oral mucosa(P0.05),but FHIT protein content was significantly associated with differentiation(P