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Hypoxia is a common challenge faced by mollusks, and the role of hypoxia-inducible factor-1α (HIF-1α) in regulating related target genes under hypoxia in Tegillarca granosa(Tg) remains unclear. In this study, we identified gene HIF-1α and further explored its function. qRT-PCR was performed to determine the mRNA expression of HIF-1α, prolyl 4-hydroxylases (PHD), hemoglobin (Hb), and myoglobin (Mb) in response to hypoxia. Dual-luciferase reporter analysis was used to assess the transcriptional activity of HIF-1α on the PHD and Hb genes during hypoxia. Results showed that the expression levels of HIF-1α and PHD mRNAs were highest in the gill and lowest in the adductor muscle. Under hypoxic conditions, HIF-1α and PHD mRNAs were both induced, and their expression levels increased significantly, peaking at 8 h and gradually decreasing thereafter. The transcription of Tg-PHD was induced by hypoxia and was Tg-HIF-1α dependent. Notably, the expression of Hb decreased in hemocytes (p < 0.01) after 8 h at 0.5 mg/L, while the expression of MbI and MbII increased (p < 0.01) in the hepatopancreas after 24 h and 120 h, respectively. Moreover, Tg-HIF-1α could transactivate the PHD promoter but not that of Hb. These findings provide valuable insights into the regulatory role of Tg-HIF-1α on Tg-PHD, safeguarding it from degradation by PHD and offering significant contributions to the understanding of HIF-1α’s function. Nonetheless, the expression of Tg-HIF-1α protein was not detected under hypoxic conditions, and additional investigations are required to quantify Tg-HIF-1α protein dynamics and establish the correlation between mRNA expression and protein levels.

Blood clams differ from their molluscan kins by exhibiting a unique red-blood (RB) phenotype; however, the genetic basis and biochemical machinery subserving this evolutionary innovation remain unclear. As a fundamental step toward resolving this mystery, we presented the first chromosome-level genome and comprehensive transcriptomes of the blood clam Tegillarca granosa for an integrated genomic, evolutionary, and functional analyses of clam RB phenotype. We identified blood clam-specific and expanded gene families, as well as gene pathways that are of RB relevant. Clam-specific RB-related hemoglobins (Hbs) showed close phylogenetic relationships with myoglobins (Mbs) of blood clam and other molluscs without the RB phenotype, indicating that clam-specific Hbs were likely evolutionarily derived from the Mb lineage. Strikingly, similar to vertebrate Hbs, blood clam Hbs were present in a form of gene cluster. Despite the convergent evolution of Hb clusters in blood clam and vertebrates, their Hb clusters may have originated from a single ancestral Mb-like gene as evidenced by gene phylogeny and synteny analysis. A full suite of enzyme-encoding genes for heme synthesis was identified in blood clam, with prominent expression in hemolymph and resembling those in vertebrates, suggesting a convergence of both RB-related Hb and heme functions in vertebrates and blood clam. RNA interference experiments confirmed the functional roles of Hbs and key enzyme of heme synthesis in the maintenance of clam RB phenotype. The high-quality genome assembly and comprehensive transcriptomes presented herein serve new genomic resources for the super-diverse phylum Mollusca, and provide deep insights into the origin and evolution of invertebrate RB.

Peptides from oyster hydrolysate (OPs) have a variety of biological activities. However, its protective effect and exact mechanism on testicular injury remain poorly understood. This study aimed to evaluate the protective effect of OPs on triptolide (TP)-induced testis damage and spermatogenesis dysfunction and investigate its underlying mechanism. In this work, the TP-induced testis injury model was created while OPs were gavaged in mice for 4 weeks. The results showed that OPs significantly improved the sperm count and motility of mice, and alleviated the seminiferous tubule injury. Further study showed that OPs decreased malonaldehyde (MDA) level and increased antioxidant enzyme (SOD and GPH-Px) activities, attenuating oxidative stress and thereby reducing the number of apoptotic cells in the testis. In addition, OPs improved the activities of enzymes (LDH, ALP and ACP) related to energy metabolism in the testis and restored the serum hormone level of mice to normal. Furthermore, OPs promoted the expression of Nrf2 protein, and then increased the expression of antioxidant enzyme regulatory protein (HO-1 and NQO1) in the testis. OPs inhibited JNK phosphorylation and Bcl-2/Bax-mediated apoptosis. In conclusion, OPs have a protective effect on testicular injury and spermatogenesis disorders caused by TP, suggesting the potential protection of OPs on male reproduction.

While MPP+ may not directly activate microglia, the initial neuronal damage inflicted by the toxin may trigger microglia, possibly leading to synergistic pro-apoptotic interaction between neuro-inflammation and toxin-induced neurotoxicity, which may further aggravate neurodegeneration. However, what molecular targets are synergistically up or downregulated during this interaction is not well understood. Here, we addressed this by co-culturing fully differentiated human SH-SY5Y cells treated with parkinsonian toxin 1-Methyl-4-phenylpyridinium (MPP+), with endotoxin-activated microglial cell line EOC 20 to determine how this interaction affects pro-apoptotic (p38, JNK, and bax:bcl2 ratios) and pro-survival (NF-κB, MEK1) signaling at both mRNA and protein levels. Concurrent MPP+ and endotoxin-treatment aggravated a decrease in SH-SY5Y cell viability and caused strong synergistic increases in the bax:bcl2 ratio, but also NF-κB and JNK signaling. These effects were attenuated by microglia inhibitor minocycline. Altogether, these data provide further molecular insights into the important role or even conditional requirement of microglia activation in the progressive neurodegenerative nature of PD.

Background: Woman with advanced-stage high-grade serous ovarian cancer (HGSOC) is likely to have a bad prognosis. Relapses are common in patients even with no evidence of disease after primary treatment. We aimed to identify the prognostic factors for disease recurrence in these patients. Methods: A nested case-control study was conducted in a large medical center in Southwest China. The primary outcome was recurrence of disease within 3 years after CR. Cox regression was used to calculate the time to event analysis in different groups.Results: Ninety- seven patients were finally included. Fifty-seven patients (58.8%) relapsed within 3 years after CR. Among all the variables, the difference in posttreatment CA-125 level was statistically significant (P ＜0.05) between the recurrent group and the progression-free group in both univariate and multivariable analysis . A cutoff value was set at the median level in the recurrent group (10 U/ml) to categorize patients into two arms. In Cox regression, the posttreatment CA-125 level was identified as a prognostic factor for recurrence with an OR of 1.05 (95% CI: 1.02-1.10, P=0.033). The median time (from initiation of treatment) until relapse was 25 months for patients whose posttreatment CA-125 levels were higher than 10 U/ml, while it was undefined for patients whose posttreatment CA-125 level were ≤ 10 U/ml. Patients with a higer posttreatment CA-125 level showed a increased risk for OC relapse compared to those with a lower posttreatment CA-125 level. Furthermore, as shown in line graphs recording serum CA-125 levels during follow-up in each recurrent case, the increments of serum CA-125 levels were delayed in recurrent OC patients who had a posttreatment CA125 level ≤ 10 U/ml compared with those with a higher CA-125 level. Conclusion: A low serum CA-125 level after primary treatment was a potential prognostic factor in women with advanced HGSOC.

In marine ecosystems, dissolved oxygen (DO) is essential for maintaining intracellular energy balance during aerobic metabolism. Bivalve mollusks are frequently exposed to hypoxia environments due to tides, temperature changes, and anthropogenic activities. The blood clam, Tegillarca granosa, mainly inhabits intertidal mudflats and is more susceptible to low oxygen events. In this study, we investigated the effect of hypoxia on immune responses in clams, and showed that hypoxia exposure reduced total hemocyte counts (THC), hemoglobin concentrations, and intracellular reactive oxygen species (ROS) levels. Also, phagocytic and cell activities of hemocyte were significantly inhibited. Furthermore, immune-related gene expression was also down-regulated. In conclusion, hypoxia greatly affected immune functions in blood clams, and our research provided the foundation for further mechanistic studies on hypoxia tolerance in clams.

Increased proinflammatory cytokines, such as interleukin (IL)-1β, may play an important role in the etiology of depression because they cause the hypothalamic–pituitary–adrenal axis to release glucocorticoids (GC) and induce dysfunction of serotonin and norepinephrine neurotransmission. Sustained increase in GC may activate microglia to induce neuroinflammation, and suppress astrocytes to produce neurotrophins, which lead to neuronal apoptosis. Here, we tested the hypothesis that glucocorticoid receptor (GR) antagonist mifepristone (RU486) may attenuate IL-1β-induced depression-like behavior by regulating the neuroinflammation and neurotrophin functions of microglia and astrocytes. Rats received intracerebroventricular injections of IL-1β (10 ng) and/or subcutaneous injections of RU486 for 14 days. Then animal depression-like behaviors, serum corticosterone concentration, the levels of pro-inflammatory cytokines (TNF-α, IL-6), mRNA and protein expressions of CD11b, GFAP and neurotrophins (pro-BDNF, BDNF, GDNF and their receptors TrkB, p75, GFRα-1 and GFRα-2) in the amygdala were studied. Compared to controls, significantly decreased rearing score and increased defecation in the open field test, decreases in ratio of open/closed time in the elevated plus maze and in sucrose preference, while increased level of corticosterone in the serum were found in the rats administrated with IL-1β. IL-1β administration also reduced the expressions of GFAP, BDNF, GDNF and its receptor GFR-α1, but increased the expressions of CD11b, pro-BDNF, p75 and pro-inflammatory cytokines (TNF-α, IL-6) concentrations. RU486 treatment markedly attenuated these changes induced by IL-1β, except for the expressions of GFR-α1. In conclusion, RU486 may improve depression-like changes by suppressing microglia and inflammation and promoting astrocytes to restore neurotrophin function.

// Yan Yu 1, * , Bingjian Lu 2, * , Weiguo Lu 3 , Shuang Li 4 , Xiuqin Li 5 , Xinyu Wang 3 , Xiaoyun Wan 3 , Yaxia Chen 3 , Suwen Feng 3 , Yao Jia 4 , Ru Yang 4 , Fangxu Tang 4 , Xiong Li 4 , Shulan Zhang 5 , Xinyan Wang 5 , Heng Wei 5 , Zhilan Peng 6 , Lin Lu 6 , Huizhen Zhong 7 , Linjun Zhao 7 , Zhangqian Huang 8 , Lin Lin 9 , Weihong Shen 9 , Yan Lu 3, 10 , Zhu Cao 1 , Jian Zou 1 , Yuejiang Ma 1 , Xiaojing Chen 1 , Qifang Tian 3 , Shiming Lu 11 , Pengyuan Liu 10, 12 , Ding Ma 4 , Xing Xie 3 and Xiaodong Cheng 3 1 Key Laboratory of Women’s Reproductive Health of Zhejiang Province, Women’s Hospital School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 2 Department of Surgical Pathology, Women’s Hospital School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 3 Department of Gynecologic Oncology, Women’s Hospital School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 4 Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China 5 Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, Liaoning, China 6 Department of Obstetrics and Gynecology, West China Second Hospital of Sichuan University, Chengdu, Sichuan, China 7 Department of Obstetrics and Gynecology, Ningbo Women and Children’s Hospital, Ningbo, Zhejiang, China 8 Department of Obstetrics and Gynecology, Shaoxing Women and Children Hospital, Shaoxing, Zhejiang, China 9 Zhejiang University Hospital, Zhejiang University, Hangzhou, Zhejiang, China 10 Institute for Translational Medicine School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 11 Department of Clinical Laboratory, Women’s Hospital School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 12 Sir Run Run Shaw Hospital School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China * These authors have contributed equally to this work Correspondence to: Xiaodong Cheng, email: chengxd@zju.edu.cn Xing Xie, email: xiex@zju.edu.cn Ding Ma, email: dma@tjh.tjmu.edu.cn Pengyuan Liu, email: pyliu@zju.edu.cn Keywords: genomics, whole-exome sequencing, complete hydatidiform mole, pathogenesis Received: March 31, 2017 Accepted: July 29, 2017 Published: September 08, 2017 ABSTRACT Complete hydatidiform mole (CHM) is a rare pregnancy-related disease with invasive potential. The genetics underlying the sporadic form of CHM have not been addressed previously, but maternal genetic variants may be involved in biparental CHM. We performed whole-exome sequencing of 51 patients with CHM and 47 healthy women to identify genetic variants associated with CHM. In addition, candidate variants were analyzed using single base extension and Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in 199 CHM patients and 400 healthy controls. We validated candidate variants using Sanger sequencing in 250 cases and 652 controls, including 205 new controls. Two single nucleotide polymorphisms, c.G48C(p.Q16H) in ERC1 and c.G1114A(p.G372S) in KCNG4 , were associated with an increased risk of CHM ( p

Clinical evidence indicated that eicosapentaenoic acid (EPA) was more effective than docosahexaenoic acid (DHA) in depression treatment. However, possible mechanisms remain unclear. Here, a chronic unpredictable mild stress (CUMS)-induced model of depression was used to compare EPA and DHA anti-depressant effects. After EPA or DHA feeding, depression-like behavior, brain n-3/n-6 PUFAs profile, serum corticosterone and cholesterol concentration, hippocampal neurotransmitters, microglial and astrocyte related function, as well as neuronal apoptosis and survival signaling pathways were studied. EPA was more effective than DHA to ameliorate CUMS-induced body weight loss, and depression-like behaviors, such as increasing sucrose preference, shortening immobility time and increasing locomotor activity. CUMS-induced corticosterone elevation was reversed by bother fatty acids, while increased cholesterol was only reduced by EPA supplement. Lower hippocampal noradrenaline and 5-hydroxytryptamine concentrations in CUMS rats were also reversed by both EPA and DHA supplement. However, even though CUMS-induced microglial activation and associated increased IL-1&beta, were inhibited by both EPA and DHA supplement, increased IL-6 and TNF-&alpha, levels were only reduced by EPA. Compared to DHA, EPA could improve CUMS-induced suppressive astrocyte biomarkers and associated BDNF-TrkB signaling. Moreover, EPA was more effective than DHA to attenuate CUMS-induced higher hippocampal NGF, GDNF, NF-&kappa, B, p38, p75, and bax expressions, but reversed bcl-2 reduction. This study for the first time revealed the mechanisms by which EPA was more powerful than DHA in anti-inflammation, normalizing astrocyte and neurotrophin function and regulating NF-&kappa, B, p38 and apoptosis signaling. These findings reveal the different mechanisms of EPA and DHA in clinical depression treatment.

While MPP

Objective: To investigate the regulatory role of fibronectin (FN) in the formation of multicellular aggregate (MCA) in ovarian cancer SKOV3 and OVCAR-3 cells and integrin expression. Methods: The dynamic formation of MCA in SKOV3 and OVCAR-3 was determined using the liquid overlay technique in the presence or absence of FN, anti-FN, RGD peptide, control RGE. The expression of α3β1, α4β1 and α5β1 integrin in monolayer cells, MCA and FN-treated MCA were determined by flow cytometry and quantitative RT-PCR. Results: OVCAR-3 and SKOV3 MCA were formed on the 4th and 8th day and peaked on the 6th and 9th day, respectively. Treatment with different concentrations of FN, LN, type IV collagen and control RGE peptide promoted MCA growth, which was mitigated by anti-FN and RGD peptide. In comparison with monolayer cells, up-regulated α3β1, α4β1 and α5β1 expression were detected in MCA while treatment with FN in both cells. Conclusions: OVCAR-3 and SKOV3 cells had varying dynamic formation of MCA in our experimental system. FN enhanced MCA formation in both cells, which was associated with increased expression of 3β1, α4β1 and α5β1 in the MCA. Therefore, FN and these integrins may be new therapeutic targets for intervention of ovarian cancer metastasis.

The transcription factor forkhead box C1 (FOXC1) has recently been proposed as a crucial regulator of triple-negative breast cancer (TNBC) and associated with TNBC metastasis. However, the mechanism of FOXC1 in TNBC development and metastasis is elusive. In this study, overexpression of FOXC1 in MDA-MB-231 cells significantly enhanced, whereas knockdown of FOXC1 in BT549 cells significantly reduced, the capabilities of TNBC cell invasion and motility in vitro and metastasis to the lung in vivo, when compared to their respective control cells. Mechanistic studies revealed that FOXC1 increased the expression of CXC chemokine receptor-4 (CXCR4), probably through transcriptional activation. AMD3100, an inhibitor of CXCR4, could block cell migration. In a zebrafish tumor model, AMD3100 could suppress cell invasion and metastasis. In addition, overexpressing CXCR4 in FOXC1-knockdown BT549 cells increased the capabilities of TNBC cell invasion and motility. In contrast, inhibition of CXCR4 with either AMD3100 or siRNA in MDA-MB-231 cells overexpressing FOXC1 reduced the capabilities of invasion and motility. Taken together, our results reveal a potential mechanism for FOXC1-induced TNBC metastasis.

Chronic stress is the major pathogenetic factor of human anxiety and depression. Zebrafish (Danio rerio) have become a novel popular model species for neuroscience research and CNS drug discovery. The utility of zebrafish for mimicking human affective disorders is also rapidly growing. Here, we present a new zebrafish model of clinically relevant, prolonged unpredictable strong chronic stress (PUCS). The 5-week PUCS induced overt anxiety-like and motor retardation-like behaviors in adult zebrafish, also elevating whole-body cortisol and proinflammatory cytokines - interleukins IL-1β and IL-6. PUCS also elevated whole-body levels of the anti-inflammatory cytokine IL-10 and increased the density of dendritic spines in zebrafish telencephalic neurons. Chronic treatment of fish with an antidepressant fluoxetine (0.1mg/L for 8days) normalized their behavioral and endocrine phenotypes, as well as corrected stress-elevated IL-1β and IL-6 levels, similar to clinical and rodent data. The CNS expression of the bdnf gene, the two genes of its receptors (trkB, p75), and the gfap gene of glia biomarker, the glial fibrillary acidic protein, was unaltered in all three groups. However, PUCS elevated whole-body BDNF levels and the telencephalic dendritic spine density (which were corrected by fluoxetine), thereby somewhat differing from the effects of chronic stress in rodents. Together, these findings support zebrafish as a useful in-vivo model of chronic stress, also calling for further cross-species studies of both shared/overlapping and distinct neurobiological responses to chronic stress.

The aim of this study was to construct THY1 eukaryotic expression plasmid and study its effects on ovarian cancer SKOV3 cells. The gene fragment coding for THY1 was obtained from human normal ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1 (+) to construct the recombinant plasmid pcDNA3.1(+)-THY1, which was transfected into SKOV3 cells. The experimental cells were classified into three groups: SKOV3-THY1, SKOV3-Null and SKOV3. The expression of gene was measured using RT-PCR and Western blotting. The percentage of apoptotic cells and cell cycle analysis and cell proliferation were assessed by flow cytometry and MTT assay. Both SKOV3-THY1 and SKOV3-null cells were inoculated subcutaneously into nude mice to determine in vivo tumorigenicity. The gene fragment of THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1 (+) and verified by PCR, restriction endonucleases digestion and DNA sequencing and the plasmid of pcDNA3.1(+)-THY1 (THY1 gene overexpression) has been stably transfected into SKOV3 cells. The analysis of flow cytometry indicated that the pcDNA3.1(+)-THY1 transfected cells in G1 phase were significantly elevated, but in S phase were decreased. The growth of transfected cells was suppressed, and more apoptosis cells were identified in pcDNA3.1(+)-THY1 transfectants compared with vector vehicle transfectants. The tumor suppressing activity of THY1 in SKOV3 cells was associated with inhibition of SKOV3 cellular proliferation, in vivo tumorigenesis in nude mice. THY1 transfection can inhibit the growth of SKOV-3 cells in vitro and in vivo. THY1 gene may play an important role in generation and development of ovarian cancers.

Background: Adenosarcoma of the uterus is one of the rare types of gynaecological malignant tumours. Poor awareness of it among clinicians makes it difficult to diagnose correctly and timely, so it is helpful to enrich and update our knowledge about the tumour with new information of patients. Aim: To improve the level of diagnosis and treatment of Mullerian adenosarcoma of the uterus. Methods: The medical data of nine patients with Mullerian adenosarcoma of the uterus who were treated from May 1995 to March 2006 in our hospital were analysed retrospectively. The analysis focused on clinicopathological features, treatment and prognosis. Results: Patients typically presented with abnormal uterine bleeding, pain in the lower abdomen, enlargement of the uterus, a mass in the uterine cavity and/or a cervical neoplasm. Microscopically, the glands were lined by benign or atypical glandular epithelium, together with sarcomatous stromal cells which showed characteristic structures of ‘periglandular cuff’ of increased cellularity and ‘intraglandular polypoid projections’. The primary diagnostic rate was 33.3% and the average interval from symptom onset to final diagnosis was 13 months and eight weeks for pre- and postmenopausal patients, respectively. The prognosis was better in early stage disease in young patients. Conclusions: The most common symptom of adenosarcoma of the uterus is abnormal uterine bleeding. Younger patients are more often misdiagnosed. Clinical stage and age of the patient are closely related to prognosis. Surgery is the primary treatment, and chemotherapy may be somewhat beneficial.

Objective To evaluate the relationship between the pelvic and para-aortic lymphadenectomy and the prognosis of epithelial ovarian cancer.

To explore an ideal approach for detecting the physical status of HPV-16 in clinic use and to investigate the integrated HPV-16 in CINs and cervical cancer. Multiplex real-time PCR method was established to quantify the copy numbers of E2 and E6 genes (E2/E6) for analysis of the physical status of HPV-16 DNA and this assay was compared to Southern blot analysis. HPV-16-containing paraffin-embedded tissues including 49 CINs and 51 cervical squamous cancers were detected using the method. (1) The cutoff ratio of E2/E6 to distinguish pure episomal from mixed HPV-16, was 0.81 in the multiplex real-time PCR; (2) The agreement rate between multiplex real-time PCR and Southern blot was 81.5% (the Kappa statistic was 0.844, P 70%) constituted the majority in CIN II and CIN III; HPV-16 DNA mostly integrated into the host chromosome (s) in squamous cervical cancers (68.6%); (4) The incidence of HPV-16 integration was increased with the degree of cervical lesions; (5) The frequency of pure integrated HPV-16 in stage II+III (88%) was significantly higher than that in stage I (33.3%). (1) Mutiplex real-time PCR provides a rapid, sensitive and reliable method for clinic detection of the physical state of HPV-16 DNA; (2) The integration of the HPV-16 DNA is a very early and important event in the progression from preinvasive to invasive cervical cancer; (3) The pure integrated status of HPV-16 in cervical cancer may be associated with poor prognosis of cervical cancer, but further study will be needed to prove its prognostic significance.

Objection To evaluate the impact of fertility-preserving surgery and adjuvant chemotherapy on survival and fertility of young patients with ovarian malignant tumors.

The balance between cell proliferation and apoptosis is critical for normal development and for the maintenance of homeostasis in adult organisms. Disruption of this balance has been implicated in a large number of disease processes, ranging from autoimmunity and neurodegenerative disorders to cancer. The ubiquitin-proteasome pathway, responsible for mediating the majority of intracellular proteolysis, plays a crucial role in the regulation of many normal cellular processes, including the cell cycle, differentiation and apoptosis. Apoptosis in cancer cells is closely connected with the activity of ubiquitin-proteasome pathway. The peptide-aldehyde proteasome inhibitor MG132 (carbobenzoxyl-L-leucyl-L-leucyl-L-leucine) induces the apoptosis of cells by a different intermediary pathway. Although the pathway of induction of apoptosis is different, it plays a crucial role in anti-tumor treatment. There are many cancer-related molecules in which the protein levels present in cells are regulated by a proteasomal pathway; for example, tumor inhibitors (P53, E2A, c-Myc, c-Jun, c-Fos), transcription factors (transcription factor nuclear factor-kappa B, IκBα, HIFI, YYI, ICER), cell cycle proteins (cyclin A and B, P27, P21, IAP1/3), MG132 induces cell apoptosis through formation of reactive oxygen species or the upregulation and downregulation of these factors, which is ultimately dependent upon the activation of the caspase family of cysteine proteases. In this article we review the mechanism of the induction of apoptosis in order to provide information required for research.

Background: Reproductive tract sarcomas metastasizing to the breast are uncommon. To our knowledge, metastasis of vaginal leiomyosarcoma to the breast has not been previously reported in the literature. Case Report: We present the first report of a FIGO stage IV primary vaginal leiomyosarcoma with metastases to the lung and left breast. Treatment included neoadjuvant chemotherapy followed by surgery and postoperative chemotherapy. Lung metastasis disappeared but recurred 14 months later in conjunction with left breast metastasis which was resected. Conclusion: Primary vaginal sarcoma with lung and breast metastases is very rare in female genital malignancies. We present this case to alert gynecologists to the need for early diagnosis and aggressive management.

Malignant granular cell tumor is relatively uncommon, constituting only 1-2% of all granular cell tumors. It is a rare and unusual tumor, especially in non-typical sites, such as the uterine cervix, and grows more rapidly than benign granular cell tumor. It can be treated with surgical excision, but recurrence is possible and prognosis can be poor. A malignant granular cell tumor in the uterine cervix of a 37-year-old woman was incidentally diagnosed. The patient has a history of irregular vaginal bleeding. Uterine cervical biopsy under colposcope revealed a malignant granular cell tumor. After isophosphamide, etoposide, and cisplatin neoadjuvant chemotherapy, surgery was performed on the lesion, which approximately involved half the depth of cervical stroma. Computed tomography examination showed no local recurrence or distant metastasis during the 26-month follow-up period.

Primary vaginal carcinoma is a rare gynecologic cancer. Radiotherapy is the standard treatment for patients affected by International Federation of Gynecology and Obstetrics stage II vaginal cancer. Neoadjuvant chemotherapy followed by radical surgery has been shown to be a valid therapeutic strategy in patients with cervical cancer; however, there is little information concerning the feasibility of neoadjuvant chemotherapy followed by radical surgery in patients with invasive vaginal carcinoma. We report the first case of stage II vaginal carcinoma with neoadjuvant chemotherapy followed by radical surgery combined with vaginal reconstruction using bilateral pudendal thigh fasciocutaneous flaps. The patient was free of disease with a satisfactory sexual life after 30 months.

An outbreak of disease on the cultured Miichthys miiuy occurred in Zhoushan of Zhejiang province. The symptom displayed as skin ulceration and the inside apparatus turned white.We isolated a dominant bacterial strain from the diseased Miichthys and assigned it as strain 090212. The artificial infection test proved that the isolate 090212 was the pathogenic bacterium that caused the disease. We applied physiological and biochemical characterization and API system in the bacterial classification. In order to confirm the result, we amplified a 1458bp sequence of 090212' s 16S rDNA and compared with other Vibrio in GenBank.The results turned out that 090212 was Gram negative and short rod with single polar flagellum. Homology analysis and phylogenetic study showed that strain 090212 had the highest similarity to Vibrio harveyi, with 99% identity. The sensitivity test of strain 090212 to 28 kinds of antibiotics revealed that the pathogen was sensitive to drugs such as Florfenicol and Tetracycline.This paper revealed for the first time that the causative pathogen, Vibrio harveyi, lead to the mass mortality of Miichthys, which will be helpful in the disease control and health management during Miichthys cultivation.

This study was designed to construct THY1 eukaryotic expression plasmid and assess its effects on epithelial ovarian cancer cell line SKOV3. The gene fragment coding for THY1 was inserted into pcDNA3.1(+) for constructing the recombinant plasmid pcDNA3.1(+)-THY1. The eukaryotic expression plasmid was analyzed by PCR, restriction endonucleases digestion and DNA sequencing. The recombinant plasmid pcDNA3.1(+)-THY1 was transfected into SKOV3 cells by liposome protocol. The experimental cells were classified into three groups: SKOV3-THY1, SKOV3-Null and SKOV3. The pcDNA3.1(+)-THY1 has been transfected into SKOV3 cells by RT-PCR and immunohistochemistry. The cell inhibitory rate of SKOV3-THY1 (56.6% at the fifth day) was higher than that of SKOV3-Null (12.5%), there was significant difference between them (P0.05). The ratios of G1 phase of SKOV3 cells after transfection were increased and the ratios of S phase were decreased significantly. There was significant difference between SKOV3-THY1 and SKOV3-Null or SKOV3 (P0.05), but there was no significant difference between SKOV3-Null and SKOV3 (P0.05). We have constructed the recombinant plasmid pcDNA3.1(+)-THY1 sucessfully. THY1 transfection can inhibit the growth of SKOV3 cells in vitro.

Bone metastasis of endometrial carcinoma is uncommon, and bilateral femur metastasis is extremely rare. A 48-year-old woman with Federation International of Gynecology and Obstetrics stage IIB grade 2-3 endometrial adenocarcinoma underwent curative radical hysterectomy, bilateral salpingo-oophorectomy, and pelvic and para-aortic lymphadenectomy followed by radiotherapy and chemotherapy. Twenty-two months after surgery, she suffered from progressive pain and then presented bilateral femur metastasis. She was administered the surgical excision of bony metastasis and adjuvant therapies including chemotherapy, palliative radiation, and hormone therapy. Three and a half years after treatment of bony metastasis, she remains clinically well. Bone metastasis of endometrial carcinoma may occur at some unusual sites, and bilateral femur metastasis should be considered in patients. Multimodal therapies are usually advocated for bone metastasis of endometrial carcinoma.

This study was aimed to explore a simple and applicable method of separating mature sperms from human testicular tissue for intracytoplasmic sperm injection and embryo transfer. The suspension of human testicular tissue was cultured in 10% human serum albumin and human tubule fluid with different concentrations (0 u/ml; 50 u/ml; 100 u/ml; 150 u/ml; 200 u/ml) of hyaluronidase for 24 h, and then the Percoll gradient centrifugation was processed to separate the sperms; meanwhile the sperms were counted and graded according to their motility. The difference in quality and quantity among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treated groups contained large quantity and high quality of sperms as compared with the two contrast groups (P0.01). The groups in the solution of 50 u/ml, 100 u/ml and 150 u/ml concentrations of hyaluronidase had almost the same amount of sperms that displayed higher motility as compared against the sperms in the group treated with 200 u/ml concentration of hyaluronidase (P0.01). There was no difference between the two contrast groups (P0.05), or among the groups treated with 50 u/ml, 100 u/ml, and 150 u/ml of hyaluronidase concentration (P0.05). This method of adopting hyaluronidase with Percoll gradient centrifugation in the process for separating mature sperms from human testicular tissue is applicable. It can increase the quantity and quality of sperms separated from testicular tissue suspensions when adequate concentrations of hyaluronidase is used.

To study the value of adenovirus mediated expression of the Escherichia coli cytosine deaminase (CD) gene with 5-fluorocytosine (5-FC) in the gene therapy of ovarian cancer cells.Ovarian cancer cells were transduced CD gene by adenovirus vector and exposed to varying concentration of 5-FC. After 5 days of incubation, MTT assays were performed to measure the cell viability.It was shown that tumor cells transduced with CD gene had a high sensitivity to 5-FC. Mixed cellular assay revealed that CD-postive cells had a neighbor cell killing effect on CD-negative cells when exposed to 5-FC. 20% CD positive cells killed more than 80% mixed cells.CD/5-FC gene therapy approach is an effective anti-tumor strategy in the treatment of ovarian cancer.

To investigate the expression and correlation of C-met and Estrogen receptor (ER) in endometrial carcinomas.We examined 55 samples of endometrial carcinomas using specific polyclonal endometrial carcinomas.antibody and monoclonal antibody correspondent to the human C-met and ER respectively. The histochemical score was estimated according to the formula H = (i + 1) x pi, with a criterion of equal or more than 70 as considered to be positive in expression.The expression of C-met was correlated with both histological grade and clinical stage of endometrial carcinomas. The H scores of carcinomas in grade 3 was higher than in grade 1 and grade 2, while higher H scores were found in carcinomas in stage III and II compared to stage I (P0.05). The higher of the grade and stage were, the weaker the ER were expressed in endometrial carcinomas (P0.05). There was an inverse correlation between the expression of ER and C-met (r = -0.5842 P0.05).The expression of C-met and ER might be a useful predictor for the prognosis of endometrial carcinomas and an indicator to evaluate the effectiveness of the hormone intervention.

To investigate whether the monoclonal antibody of basic fibroblast growth factor (bFGF-MAb) inhibits the proliferation, invasion and angiogenesis of ovarian cancer.1. Human ovarian cancer cells SKOV3 were planted in 24-well dishes, to which was added bFGF-MAb at different concentrations, and crystal violet staining was performed daily for 8 days, then cell numbers were counted by OD490 determination. 2. SKOV3 were transplanted intraperitoneally to BALB/c nude mice. One week later, bFGF-MAb was injected intraperitoneally twice a week, and the survival time of nude mice, the number and weight of the metastatic tumors on the mensentery were observed or measured. 3. Intratumoral microvessel density(MVD) was measured by immunohistochemical staining for CD31.1. bFGF-MAb inhibited the proliferation of SKOV3 in the concentration-dependent manner (P0.05). 2. The average survival time of nude mice in bFGF-MAb group increased by 10 days over that of control. 3. The average number and weight of metastatic tumors on mensentery in bFGF-MAb group constituted 70.6% and 69.2% of those in control group, respectively. 4. The MVD in bFGF-MAb group accounted for 62.8% of the MVD in control group.bFGF-MAb can inhibit the proliferation and angiogenesis of ovarian cancer markedly, so it promises to have application in the biological treatment of ovarian cancer.

To assess the efficacy of microdetermination in the ATP-bioluminescence assay (ATP-CVA) of ovarian cancer chemosensitivity testing in vitro.A micromethod ATP-CVA was employed in determining the chemosensitivity of 35 specimens of fresh ovarian cancer cells using Victor 2 Multilabel Counter.1. By use of Victor 2 Multilabel Counter to determine ATP, in a range of 10(-9)-10(-5) mol/ml, a linear relationship was remarkable between the log ATP and the log luminescence. The equation was Y = 0.892X + 10.257, the correlation coefficient r was 0.998, P0.01. 2. The coefficient of variation obtained from ATP-CVA was 7.3%. 3. The sensitivity of the test was 92.0%, the specificity 70.0%, and the total predictive accuracy of ATP-CVA 87.1%.With the use of Victor 2 Multilabel Counter, ATP-CVA is fast, cost-effective, and easy to perform with only small error.

To evaluate the significance of applying short tandem repeat DNA in the model of human-mouse cord blood transplantation.The techniques of STR-PCR and colony forming cell culture were used to examine the engrafting and influence of hematopoietic growth factor on process in the mice after transplantation of human cord blood (CB).1. Human DNA expression was detected in the mice that had received tail-vein injection of human CB (1-5 x 10(7) MNC per mouse) but it was not detected in the mice that have received intraperitoneal injection of human CB. 2. Human CB transplanted in SCID mice could reconstitute the hematopoietic function of the mice; the expression level of human cell was not related with the use of hematopoietic growth factor.STR-PCR analysis provide a simple and accurate indication of engraftment in the model of human-mouse cord blood transplantation.

To investigate the effects of human wild-type p53 gene transfection on the growth and chemosensitivity of human cervical carcinoma.Recombinant eukaryotic expression vector pCB6.p53 containing human wild-type p53 cDNA was introduced by lipofectamine transfection regent into HeLa cell line. The expression of p53 was detected by immunohistochemistry. Cell proliferation was measured by methyl thiazolyl tetrazolium after the treatment of cisplatin.Expression of p53 was detected by immunohistochemistry. The growth rate of p53-transfected HeLa decreased. Wild-type p53-positive HeLa was more sensitive to cisplatin, compared with the control cell lines.The exogenous wild-type p53 expression not only induced great suppression of cell growth but also increased chemosensitivity of human cervical carcinoma.

To assess the effect of progesterone on the gene expression of insulin-like growth factor-I receptor (IGF-I R) in human decidual stromal cells of early pregnancy in vitro.Semiquantitative reverse transcriptase polymerase chain reaction, using beta-ACTIN as internal standard, was applied to determine the levels of IGF-I R mRNA in human decidual stromal cells of early pregnancy in vitro after cultured with different concentrations of progesterone for 72 hours or cultured with 0.1 mumol/L of progesterone for different periods of time.The expression of IGF-I R mRNA was significantly positive in human decidual stromal cells of early pregnancy in vitro. The levels of IGF-I R mRNA were down-regulated by progesterone, and showed significant negative-correlation with the concentration of progesterone (r = -0.680, P0.001). The levels of IGF-I R mRNA showed no significant correlation with time when the final concentration of progesterone was 0.1 mumol/L (r = 0.005, P0.05).Progesterone may play an important role in the regulation of proliferation and decidualization of stromal cells by down-regulating the expression of IGF-I R mRNA in human decidual stromal cells of early pregnancy, which is important for the maintenance of early pregnancy.

To investigate the gene expression of insulin-like growth factors(IGFs) in human villous trophoblast cells of early pregnancy in vitro and their action.Semiquantitative reverse transcriptase polymerase chain reaction(RT-PCR), with beta-ACTIN as internal standard, was applied to determine the expression of IGFs messenger RNA in human villous trophoblast cells of early pregnancy in vitro.The expression of IGF-II mRNA, IGF-I R mRNA and IGFBP-3 mRNA was detectable in human villous trophoblast cells of early pregnancy in vitro.IGFs appear to play important regulation roles in early invasion, proliferation and differentiation of cytotrophoblast, and in the formation of placenta and the development of embryo via autocrine and/or paracrine way.