Histamine promotes mouse decidualization through stimulating epithelial amphiregulin release.

Accumulating evidence shows that inflammation is essential for embryo implantation and decidualization. Histamine, a proinflammatory factor that is present in almost all mammalian tissues, is synthesized through decarboxylating histidine by histidine decarboxylase (HDC). Although histamine is known to be essential for decidualization, the underlying mechanism remains undefined. In the present study, histamine had no obvious direct effects on in vitro decidualization in mice. However, the obvious differences in HDC protein levels between day 4 of pregnancy and day 4 of pseudopregnancy, as well as between delayed and activated implantation, suggested that the blastocyst may be involved in regulating HDC expression. Furthermore, blastocyst-derived tumor necrosis factor α (TNFα) significantly increased HDC levels in the luminal epithelium. Histamine increased the levels of amphiregulin (AREG) and disintegrin and metalloproteinase domain-containing protein 17 (ADAM17) proteins, which was abrogated by treatment with famotidine, a specific histamine type 2 receptor (H2R) inhibitor, or by TPAI-1 (a specific inhibitor of ADAM17). Intraluminal injection of urocanic acid (HDC inhibitor) on day 4 of pregnancy significantly reduced the number of implantation sites on day 5 of pregnancy. TNFα-stimulated increases in HDC, AREG and ADAM17 protein levels was abrogated by urocanic acid, a specific inhibitor of HDC. Additionally, AREG treatment significantly promoted in vitro decidualization. Collectively, our data suggests that blastocyst-derived TNFα induces luminal epithelial histamine secretion, and histamine increases mouse decidualization through ADAM17-mediated AREG release.

Tryptophan in the mouse diet is essential for embryo implantation and decidualization.

Introduction: Nutritional deficiency occurs frequently during pregnancy and breastfeeding. Tryptophan (Trp), an essential amino acid which is critical for protein synthesis, serves as the precursor for serotonin, melatonin, and kynurenine (Kyn). The imbalance between serotonin and kynurenine pathways in Trp metabolism is closely related to inflammation and depression. This study assessed the effects of Trp deficiency on mouse early pregnancy. Methods: Embryo implantation and decidualization were analyzed after female mice had been fed diets containing 0.2% Trp (for the control group), 0.062% Trp (for the low Trp group) and 0% Trp (for the Trp-free group) for two months. The uteri of the mice were collected on days 4, 5, and 8 of pregnancy for further analysis. Results: On day 8 of pregnancy, the number of implantation sites were found to be similar between the control and the low Trp groups. However, no implantation sites were detected in the Trp-free group. On day 5 of pregnancy, plane polarity- and decidualization-related molecules showed abnormal expression pattern in the Trp-free group. On day 4 of pregnancy, there was no significant difference in uterine receptivity molecules between the low-Trp group and the control group, but uterine receptivity was abnormal in the Trp-free group. At implantation sites of the Trp-free group, IDO and AHR levels were markedly elevated. This potentially increased levels of Kyn, 2-hydroxy estradiol, and 4-hydroxy estradiol to affect decidualization. Conclusions: Trp-free diet may impair decidualization via the IDO-KYN-AHR pathway.

[Empowering Persons With Mental Illness to Begin the Recovery Journey: An Example of Overcoming Internalized Stigma].

Internalized stigma is a significant obstacle encountered on the journey to recovery for persons with mental illness. Services provided through the empowerment model of care have been shown to be effective in overcoming this obstacle. However, the mental health care system in Taiwan continues to focus on providing care using the medical or rehabilitation model. In this article, the related consequences and psychological effect mechanism of internalized stigma in persons with mental illness are presented and evidence-based interventions for internalized stigma are demonstrated. Next, we share the clinical experiences to advocate for the use by professionals of the empowerment model. Self-efficacy and empowerment may mediate the psychological effects of internalized stigma on quality of life and depression, while social networks and peer support may buffer its negative consequences. Related evidence-based interventions include psycho-education, motivation enhancement, self-help or peer-delivered services, and cognitive behavioral model and goal-oriented skill training. Empowerment model services are based on scientific evidence that are practiced under the rubric of self-determination and constructed in supportive environments. When examining psychiatric symptoms in persons with mental illness, professionals may concurrently evaluate for issues related to internalized stigma and encourage those patients with internalized stigma to self-determine and participate in therapy. In psycho-education and skill training therapies, professionals may also apply the lived experiences of peers with recovery to identify and share important knowledge and life skills. When connecting patients to community resources, professionals should encourage stakeholders to help persons with mental illness put their decisions into practice.

[Correlation between the Promoter Methylation Status of TRIM58 and Its mRNA Expression in Acute Myeloid Leukemia].

OBJECTIVE: To analyze the relationship between the promoter methylation status of Tripartite-motif protein 58 (TRIM58) and its mRNA expression level in acute myeloid leukemia (AML), and to explore the expression and regulation of TRIM58 in AML. METHODS: The bisulfite sequencing PCR (BSP) and quantitative real-time PCR (qPCR) technologies were used to detect the promoter methylation status and expression levels of TRIM58 mRNA in primary CD34+ and CD34- AML cells and the AML cell lines KG1a and K562 were determined. RESULTS: The expression of TRIM58 mRNA in CD34+ cells was down-regulated in 10 AML patients, while that in CD34- cells was down-regulated in 12 cases. Differences in the promoter methylation level of TRIM58 were statistically significant between AML group and normal control group (P<0.05). Additionally, the expression of TRIM58 mRNA was down-regulated in cell lines KG1a and K562, and up-regulated after decitabine treatment. CONCLUSION: The down-regulation of mRNA expression of TRIM58 in AML cells may be related to its promoter methylation status.

The Feasibility of a Training Program for Peers with Severe Mental Illness to Provide One-to-One Services in Taiwan: A Pilot Study.

BACKGROUND: In Taiwan, services provided by patients' peers in the mental health care system are still lacking. Therefore, this study aimed to develop a community setting model by a training program for severe mental illness (SMI) patients' peers that also have SMI in Taiwan. METHOD: This training program comprised of 13-h lectures, 15-h practice classes, and an eight-week internship. In 2018 and 2019, the trainees provided one-to-one services to service users with SMI during the internship at a halfway house. The satisfaction and outcomes among all participants were measured in this training course. RESULTS: The total mean satisfaction score in the training course for trainees (10 items, n = 13) and internship services for service users (12 items, n = 29) were 4.7 ± 0.4 and 4.6 ± 0.5, respectively. Among the trainees, 11 demonstrated improved Brief Psychiatric Rating Scale-18 (BPRS-18), Chinese Health Questionnaire-12 (CHQ-12), and Global Assessment of Functioning (GAF) scores after the whole training course. Among the 29 service users, their scores in the BPRS-18 and CHQ-12 decreased, whereas their scores in the GAF increased significantly under the internship service. CONCLUSION: In this pilot study, the trainees and service users who received internship services felt satisfied. The service providers and users with SMI both showed better clinical outcomes.

Percutaneous Abdominal Puncture for Catheterization with Assistance of Guidewire in Lumboperitoneal Shunt.

BACKGROUND: When using a classic lumboperitoneal shunt, laparotomy is inevitable for peritoneal catheter implantation, which is time consuming and difficult for unskilled neurosurgeons. A minimally invasive technique of percutaneous abdominal puncture for catheterization with the assistance of guidewire is introduced in this paper. METHODS: Ten patients with communicating hydrocephalus received a lumboperitoneal shunt through percutaneous abdominal puncture for catheterization. The safety and effectiveness of percutaneous abdominal puncture for catheterization were followed up for more than 6 months. RESULTS: The surgery was successfully completed in 10 patients. The average operation time was nearly 30 minutes. No patients reported abdominal organ damage. None of the 10 patients had other complications such as peritonitis and obstruction of abdominal catheters caused by the percutaneous peritoneal puncture technique. One patient presented with intracranial aseptic inflammation postoperatively, which was controlled after repeated lumbar puncture. Another patient confirmed that the lumbar catheter was folded in 6 months postoperatively. CONCLUSIONS: The technique of percutaneous abdominal puncture for catheterization with the assistance of a guidewire is a simple, safe, and effective way to treat communicating hydrocephalus.

The correlation between VitD3 levels and the disease activity of childhood-onset systemic lupus erythematosus.

BACKGROUND: There is growing evidence linking low levels of vitamin D3 to an increased risk of many autoimmune diseases. Compared to the general population, hypovitaminosis D is more prevalent among children with systemic lupus erythematosus (SLE), which can be associated with sun exposure avoidance, long-term corticosteroid treatment, and renal disease. Therefore, we launched this study to assess the correlation between 25 (OH) D3 (VitD3) levels and the disease activity of children with SLE (cSLE) in Taiwan. METHODS: From September to December 2018, we recruited 31 cSLE patients from the Pediatric Out-patient Department of Taichung Veterans General Hospital. Their basic data, including SLE disease index 2000 (SLEDAI-2K) score, laboratory values, prescribed drugs and VitD3 levels were collected and analyzed statistically. RESULTS: The mean serum VitD3 concentration was 19.7 ± 7.9 ng/mL and SLEDAI-2K 6.2 ± 5.0. Those patients (N = 16) with an SLEDAI-2K≦4 had higher VitD3 levels when compared to those (N = 15) with an SLEDAI-2K>4 (22.9 ± 7.7 vs 16.3 ± 6.7 points, p = 0.020). Five patients not taking systemic corticosteroids (SCS) had significantly higher VitD3 levels and lower SLEDAI-2K than those who took SCS (N = 26). Additionally, we found VitD3 levels to be negatively correlated to SLEDAI-2K (rs = -0.55, p = 0.001) and daily SCS dosages (rs = -0.49, p = 0.005). CONCLUSION: This study shows that VitD3 deficiency is common in patients with cSLE. It was also noted that serum VitD3 levels negatively correlate to SLEDAI-2K, which can be partially explained by less usage of SCS.

Mitochondrial oxidative phosphorylation is dispensable for survival of CD34+ chronic myeloid leukemia stem and progenitor cells.

Chronic myeloid leukemia (CML) are initiated and sustained by self-renewing malignant CD34+ stem cells. Extensive efforts have been made to reveal the metabolic signature of the leukemia stem/progenitor cells in genomic, transcriptomic, and metabolomic studies. However, very little proteomic investigation has been conducted and the mechanism regarding at what level the metabolic program was rewired remains poorly understood. Here, using label-free quantitative proteomic profiling, we compared the signature of CD34+ stem/progenitor cells collected from CML individuals with that of healthy donors and observed significant changes in the abundance of enzymes associated with aerobic central carbonate metabolic pathways. Specifically, CML stem/progenitor cells expressed increased tricarboxylic acid cycle (TCA) with decreased glycolytic proteins, accompanying by increased oxidative phosphorylation (OXPHOS) and decreased glycolysis activity. Administration of the well-known OXPHOS inhibitor metformin eradicated CML stem/progenitor cells and re-sensitized CD34+ CML cells to imatinib in vitro and in patient-derived tumor xenograft murine model. However, different from normal CD34+ cells, the abundance and activity of OXPHOS protein were both unexpectedly elevated with endoplasmic reticulum stress induced by metformin in CML CD34+ cells. The four major aberrantly expressed protein sets, in contrast, were downregulated by metformin in CML CD34+ cells. These data challenged the dependency of OXPHOS for CML CD34+ cell survival and underlined the novel mechanism of metformin. More importantly, it suggested a strong rationale for the use of tyrosine kinase inhibitors in combination with metformin in treating CML.

[Research Advance of Gene Mutation and Targeted Drug Therapy in Childhood Acute Myeloid Leukemia --Review].

The clinical therapeutic regimen for acute myeloid leukemia (AML) is not significantly different between adults and children, which is mostly based on IA (idarubicin and cytosine arabinoside) induction chemotherapy. With the rapid development of sequencing technique, people's understandings towards the molecular and biological abnormalities of AML are increasing, diverse AML gene mutation-based targeted drugs have been rapidly developed and applied. In this review, several commonly gene mutations in AML (such as FLT3, NPM1 and C/EBPA) was described, and the therapeutic effects and differences of targeted drugs that used in clinical treatment or had been reported (like tyrosine kinase inhibitor, IDH1 mutation inhibitor and epigenetic modification inhibitor) in child and adult AML patients were summrized.

BHPF exposure impairs mouse and human decidualization.

Although BHPF has been widely used in plastic manufacturing as a substitute for BPA, current evidence suggests that BHPF also causes harmful effects on reproduction. However, effects of BHPF on mammalian early pregnancy are still poorly defined. This study aimed to explore the effects of BHPF on early pregnancy, especially decidualization and embryonic development in mice and human beings. The results showed that 50 and 100 mg/kg BHPF exposure reduced birth weight, and implantation site weight on the day 8 of pregnancy in mice. Because BHPF inhibits both embryo development and artificial decidualization in mice, suggesting that the detrimental effects of BHPF should be from its effects on embryo development and decidualization. Under in vitro decidualization, 10 µM BHPF inhibits decidualization and leads to disordered expression of Lamin B1 and collagen in mice. In addition, 10 µM BHPF also inhibits decidualization, and causes disordered expression of both collagen III and Lamin B1 under human in vitro decidualization. However, collagen III supplementation can rescue BHPF inhibition on decidualization. Further, our study demonstrates that BHPF impairs human decidualization through the HB-EGF/EGFR/STAT3/Collagen III pathway. Taken together these data suggest that exposure to BHPF impairs mouse and human decidualization during early pregnancy.

Research Advance of Gene Mutation and Targeted Drug Therapy in Childhood Acute Myeloid Leukemia --Review / 中国实验血液学杂志

The clinical therapeutic regimen for acute myeloid leukemia (AML) is not significantly different between adults and children, which is mostly based on IA (idarubicin and cytosine arabinoside) induction chemotherapy. With the rapid development of sequencing technique, people's understandings towards the molecular and biological abnormalities of AML are increasing, diverse AML gene mutation-based targeted drugs have been rapidly developed and applied. In this review, several commonly gene mutations in AML (such as FLT3, NPM1 and C/EBPA) was described, and the therapeutic effects and differences of targeted drugs that used in clinical treatment or had been reported (like tyrosine kinase inhibitor, IDH1 mutation inhibitor and epigenetic modification inhibitor) in child and adult AML patients were summrized.

The social support, mental health, psychiatric symptoms, and functioning of persons with schizophrenia participating in peer co-delivered vocational rehabilitation: a pilot study in Taiwan.

BACKGROUND: Vocational peer support (VPS) services are recovery-oriented interventions in modern psychiatric care for persons with schizophrenia. However, few VPS services are found in Taiwan. Hence, a pilot program of peer co-delivered vocational rehabilitation to support persons with schizophrenia in Taiwan was proposed and evaluated. METHODS: Six peers were trained and were willing to co-lead and assist workplace problem-solving groups and care skills training in an extended vocational rehabilitation program from August 2017 to December 2018. The social support, mental health, psychiatric symptoms, and functioning of service users were assessed before and after peer co-delivered services, and the assessments were based on the following: Social Support Scale (SSS), Chinese Health Questionnaire-12 (CHQ-12), Brief Psychiatric Rating Scale (BPRS), Global Assessment of Function (GAF), and the Chinese version of the Social Functioning Scale (C-SFS). RESULTS: The recruited 46 service users were mostly middle-aged (49.1 ± 9.8), with 27 being male (58.7%). After interventions, 42 service users who completed the program had a significantly increased SSS score (149.1 ± 31.8 vs. 161.2 ± 35.0, df = 41, t = 2.70, p = 0.01) and subscale of friend-peer dimension (44.4 ± 12.0 vs. 53.2 ± 13.2, df = 41, t = 4.72, p < 0.001). The objective (GAF: 69.8 ± 9.8 vs. 72.6 ± 8.8, df = 41, t = 3.50, p = 0.001) and subjective social functional scores (C-SFS: 75.2 ± 8.8 vs. 78.1 ± 9.5, df = 41, t = 2.59, p = 0.01) both significantly increased. The weekly wage elevated significantly (37.5 ± 35.5 vs. 43.6 ± 38.0, df = 41, t = 2.57, p = 0.01) and the BPRS-18 score decreased significantly, too (31.2 ± 6.7 vs. 29.3 ± 5.0, df = 41, t = - 2.83, p = 0.007). CONCLUSIONS: Peer co-delivered vocational rehabilitation services may enhance the social support received by persons with schizophrenia and improve their occupational outcomes. The pilot program proposed can thus be a model for non-Western countries with limited resources allocated by governments to support persons with schizophrenia. TRIAL REGISTRATION: ClinicalTrials NCT04767204, retrospectively registered on Feb 23, 2021.

Effects of pargyline on histone methylation in promoter and enhancer regions and transcription of CYP3A4/3A7 / 药学学报

This study was designed to investigate effects of pargyline on histone methylation in the promoter and enhancer regions and transcription of cytochrome P450 3A4/3A7 (CYP3A4/3A7) gene. Human primary fetal liver cells were isolated, cultured and randomly divided into several groups including control, solvent, pargyline low, middle, high dose (treated with 0.6, 1.2, 2.4 mmol·L-1). HepG2 cells were cultured and treated with 0.03, 0.3, 3 mmol·L-1 pargyline. After 48 hours, total RNAs were prepared from the cells to determine the expression of CYP3A mRNA in primary fetal cells and HepG2 cells with real-time quantative PCR (qPCR). HepG2 cells were cultured and then treated with 3 mmol·L-1 pargyline for 48 hours. The chromatin immunoprecipitation (ChIP) assay was performed with dimethylation of histone H3 at lysine 4 (H3K4me2), and IgG antibodies respectively. The precipitated DNA was resuspended and used for qPCR. Primers were used to detect different regions of CYP3A4/3A7 promoter and enhancer. Occupancy of H3K4me2 was shown as percent of input DNA relative to control cells. The results suggested that pargyline has an effect on primary fetal liver cells and HepG2 cells proliferation. The level of CYP3A7 was markedly enhanced in human primary fetal liver cells by treatment with 1.2, 2.4 mmol·L-1 of pargyline (P-1 of pargyline in HepG2 cells (P<0.001) compared with solvent control. Occupancy of H3K4me2 on human CYP3A4 promoter (-362 to +53) and enhancer segment (-7 836 to -6 093) harbored the overlapping hepatocyte nuclear factors 4A (HNF4A) binding site compared with a negative control. Occupancy of H3K4me2 on human CYP3A7 promoter (-163 to +103) and enhancer segment (-4 054 to -3 421, -6 265 to -6 247) overlapped with glucocorticoid receptor (GR) binding site. In conclusion, the enriched H3K4me2 in the promoter and enhancer regions was induced by pargyline with HNF4A or GR binding site in CYP3A4/3A7 gene to activate the corresponding genes.

Mortality among patients with schizophrenia and vocational rehabilitation program services under Taiwan's psychiatric care reform.

BACKGROUND: Vocational rehabilitation programs are implemented to enhance the occupational functioning of long-stay patients with schizophrenia. Unemployment is associated with a higher risk of death. Schizophrenia patients who participate in vocational rehabilitation programs may have better health outcomes with participation in employment. AIM: To evaluate the relationship between mortality among schizophrenia patients and vocational rehabilitation program services under Taiwan's psychiatric care reform. METHODS: A total of 2457 long-stay schizophrenia patients were followed-up retrospectively from 1998 to 2008 at Taipei Veterans General Hospital Yuli Branch in Taiwan. We collected data on annual measurements of effectiveness and the human resources utilized in the vocational rehabilitation program. Pearson's correlations between the above-collected data and the crude death rates for all patients were examined. We also assessed the association between participation in supported or sheltered employment and death. RESULTS: Most of the patients were male (81.3 %). The mean ± SD age of the patients was 57.8 ± 17.0 years. The annual crude death rate averaged 5.3 %. Both the number of community workplaces and the total wages earned from sheltered and supported employment had significantly negative linear correlations with the crude death rate among all patients (both γ ≤ -0.64, p < 0.05). After controlling the confounding factors, participation in supported or sheltered employment was significantly associated with a lower risk of death (n = 2174, HR = 0.22, 95 % CI 0.16-0.29). CONCLUSIONS: Under psychiatric care reform, the vocational rehabilitation program was more effective and there was less patient mortality. Patients who had experienced sheltered or supported employment had a lower risk of death than those who had not.

Mortality among long-stay patients with schizophrenia during the setting-up of community facilities under the Yuli model.

Objective: Over the past 15 years, Yuli Veterans Hospital (YVH) in Taiwan has developed the Yuli model to reform long-stay care for psychiatric patients. The development of the Yuli model could be divided into pre-early (1998-1999), early (2000-2006) and late (2007-2008) periods according to the setting-up of the community facilities. In the pre-early period, a vocational rehabilitation program was established for psychiatric patients in YVH. In the later periods, the independent living skills training and the program for social reintegration were instituted in the community facilities. This study aimed to evaluate mortality among the long-stay patients with schizophrenia during the three periods. Methods: In all, 2457 patients with schizophrenia who had been hospitalized for at least one year initially were retrospectively followed from 1 January 1998 to 31 December 2008. Compared with the general population in Taiwan, we calculated the age- and sex-specific standardized mortality ratios (SMRs) of those patients by cause of death during the three periods. Results: Most of the patients were male (81.3%). The mean ± SD age of the patients was 57.83 ± 16.95 years. The all-, natural- and unnatural-cause mortalities of the patients were nearly two times greater than those of the general population during the whole study period. Compared with those in the pre-early and early periods, all patients in the late period had the lowest mortality gaps. In the pre-early, early and late periods, the all-cause SMR were 5.40 (95% confidence interval (CI) = 4.27-6.81), 2.90 (95% CI = 2.20-3.79) and 1.17 (95% CI = 0.54-2.22), respectively, for the 50-69-year-old male patients. Nearly half of all the patients who participated the whole comprehensive rehabilitation program belonged to this sex and age group (N = 156, 46.6%). Conclusions: With the setting-up of community facilities for the comprehensive rehabilitation program, the mortality gaps among the 50-69-year-old male patients apparently decreased using the Yuli model.

Effect of FK506 nanospheres on regeneration of allogeneic nerve after transplant

OBJECTIVE@#To discuss effect of FK506 nanospheres used at different time on the regeneration of allogeneic nerve after transplant.@*METHODS@#Single emulsion-solvent evaporation method (O/W) was adopted to prepare the FK506 nanospheres and the tibial nerve of rats after allogeneic transplantation. FK506 nanospheres were used in group A after operation immediately, in group B in 24 h after operation, and in group C in 3 d after operation while FK506 nanospheres were not used in group D; in the 4th, 8th and 12th week after operation respectively, general observation of transplanted nerves, histological examination, image analysis of myelinated fibers, wet-weight determination of musculi triceps surae, retrogradely labeling of neurons by the fluorescein and electrophysiological comparison of bilateral tibial nerve were carried out.@*RESULTS@#FK506 nanospheres can be degraded and absorbed quickly. The neural regenerations in group A and B were similar, which were both much better than those in group C and D. The difference was statistically significant and so was the difference between group C and D.@*CONCLUSIONS@#Drug release rate of FK506 nanospheres is accordant with the regeneration law of damaged nerves and the local application can promote the regenerations of nerves. The effect would be better if the drug is used in earlier period (within 24 h).

Over-expression of testis-specific expressed gene 1 attenuates the proliferation and induces apoptosis of GC-1spg cells / 华中科技大学学报（医学）（英德文版）

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

Over-expression of testis-specific expressed gene 1 attenuates the proliferation and induces apoptosis of GC-1spg cells / 华中科技大学学报（医学）（英德文版）

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

Correlation between cell attachment areas after 2 h of culture and osteogenic differentiation activity of rat mesenchymal stem cells on hydroxyapatite substrates with various surface properties.

The initial attachment of mesenchymal stem cells (MSCs) to substrates and osteogenic differentiation are supported by culture on a hydroxyapatite substrate. Cell attachment areas of rat MSCs after 2 h of culture on hydroxyapatite substrates with various microstructures and the osteogenic differentiation activity thereafter were measured. The perceived outcome was that, after 2 h of culture, rat MSCs with a small attachment area would have a high osteogenic differentiation activity, whereas those with a large attachment area would have a low osteogenic differentiation activity. Furthermore, rat MSCs with a small attachment area had many cytoplasmic processes, while those with a large attachment area revealed clear stress fibers and focal contacts. These results suggest that cell attachment area of rat MSCs after 2 h of culture has a strong effect on the osteogenic differentiation of rat MSCs. Thus, the measurement of cell attachment area after 2 h of culture could become valuable for estimating the osteogenic differentiation activity of rat MSCs thereafter.