Effectiveness and safety of REBACIN as a non-invasive intervention for persistent high-risk human papillomavirus infection: A real-world prospective multicenter cohort study.

BACKGROUND: We previously reported that REBACIN effectively eliminates persistent high-risk human papillomavirus (hrHPV) infection. Here, we conducted a prospective multicenter cohort study to evaluate the safety and effectiveness of REBACIN, taking into account factors such as specific hrHPV subtype and patient's age. METHODS: According to inclusion/exclusion criteria and participant willingness, 3252 patients were divided into REBACIN group while 249 patients into control group. Patients in REBACIN group received one course treatment of intravaginal administration of REBACIN while no treatment in control group. After drug withdrawal, participants in both groups were followed up. RESULTS: The clearance rate of persistent hrHPV infection in REBACIN group was 60.64%, compared to 20.08% in control group. Specifically, the clearance rates for single-type infection of HPV16 or HPV18 were 70.62% and 69.23%, respectively, which was higher than that of HPV52 (59.04%) or HPV58 (62.64%). In addition, the single, double, and triple/triple+ infections had a clearance rate of 65.70%, 53.31%, and 38.30%, respectively. Moreover, 1635 patients under 40 years old had a clearance rate of 65.14%, while it was 55.08% for 1447 patients over 40 years old. No serious adverse effects were found. CONCLUSION: This study confirmed that REBACIN can effectively and safely eliminate persistent hrHPV infection, which the clearance rate of HPV16/18 is higher than that of HPV52/58, the clearance rate of single-type infection is higher than that of multiple-type infections, and the clearance rate in young patients is higher than that in elder patients, providing a guidance for REBACIN application in clearing hrHPV persistent infection in real-world settings. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry Registration Number: ChiCTR1800015617 http://www.chictr.org.cn/showproj.aspx?proj=26529 Date of Registration: 2018-04-11.

Correlations between alterations of T-helper 17 cells and treatment efficacy after concurrent radiochemotherapy in locally advanced cervical cancer (stage IIB-IIIB): a 3-year prospective study.

BACKGROUND: Recently, T-helper 17 (Th17) cells have been proved to play an important role in promoting cervical cancer. But, till now, few study has been carried out to understand the involvement of these cells in efficacy of anti-tumor treatments. This study aimed to investigate the alterations in the percentage of circulating Th17 cells and related cytokines in locally advanced cervical cancer (LACC) patients before and after concurrent chemoradiotherapy (cCRT) and to analyze the correlations between the alterations in Th17 cells and treatment efficacy. METHODS: A prospective study with 49 LACC (International federation of gynecology and obstetrics [FIGO] stage IIB-IIIB) patients and 23 controls was conducted. Patients received the same cCRT schedule and were followed up for 3 years. Circulating Th17 cells (CD3+CD8- interleukin [IL]-17+ T cells) and related cytokines IL-17, transforming growth factor-ß (TGF-ß), IL-10, IL-23, IL-6, and IL-22 were detected before and after cCRT. Correlations between alterations of circulating Th17 cells and treatment efficacy were analyzed. Kaplan-Meier analysis was used for overall survival (OS) and progression-free survival (PFS). RESULTS: We found that 40 patients finished the entire cCRT schedule and met the endpoint of this study. The percentage of circulating Th17 cells in the LACC patients was higher than that in the controls, and it significantly decreased after cCRT (P < 0.05). After cCRT, patients were divided into two groups based on the average of the Th17 cells declined. The subgroup of patients with a prominent decrease in circulating Th17 cells after cCRT had a higher treatment efficacy and longer PFS and OS times. Compared with the control patients, LACC patients had higher IL-6, IL-10, IL-22, TGF-ß levels and a lower IL-23 level (P < 0.05). After cCRT, IL-6, IL-10, IL-17, IL-23 level significantly increased and TGF-ß level significantly decreased compared with the levels before cCRT (P < 0.05). CONCLUSION: Circulating Th17 cells in the LACC patients (FIGO stage IIB-IIIB) were higher than those in the controls, but they generally decreased after cCRT. A more pronounced decrease in circulating Th17 cells after cCRT was correlated with better therapeutic effect and longer PFS and OS times.

Peritoneal cancer after bilateral mastectomy, hysterectomy, and bilateral salpingo-oophorectomy with a poor prognosis: A case report and review of the literature.

BACKGROUND: Primary peritoneal cancer (PPC) patients with BRCA mutations have a good prognosis; however, for patients with BRCA mutations who are diagnosed with PPC after prophylactic salpingo-oophorectomy (PSO), the prognosis is poor, and survival information is scarce. CASE SUMMARY: We treated a 56-year-old woman with PPC after bilateral mastectomy, hysterectomy, and bilateral salpingo-oophorectomy. This patient had primary drug resistance and died 12 mo after the diagnosis of PPC. The genetic test performed on this patient indicated the presence of a germline BRCA1 mutation. We searched the PubMed, Scopus, and Cochrane databases and extracted studies of patients with BRCA mutations who developed PPC after PSO. After a detailed literature search, we found 30 cases, 7 of which had a history of breast cancer, 14 of which had no history of breast cancer, and 9 of which had an unknown history. The average age of PSO patients was 48.86 years old (range, 31-64 years). The average time interval between the diagnosis of PPC and preventive surgery was 61.03 mo (range, 12-292 mo). The 2-year survival rate for this patient population was 78.26% (18/23), the 3-year survival rate was 50.00% (9/18), and the 5-year survival rate was 6.25% (1/16). CONCLUSION: Patients with BRCA mutations who are diagnosed with PPC after preventative surgery have a poor prognosis. Prevention measures and treatments for these patients need more attention.

Acquired uterine arteriovenous fistula due to a previous cornual pregnancy with placenta accreta: A case report.

The present report describes a case of an acquired uterine arteriovenous fistula, which, following surgery and postoperative pathological analysis, was confirmed as a previous cornual pregnancy with placenta accreta. The patient was a 37-year-old woman (gravida 3; para 2) who had previously delivered two children via cesarean section (in 2004 and 2010, respectively) and also had a spontaneous abortion (2008). She had experienced continuous menstrual bleeding for ~20 days and had a history of prolonged (~30 days) vaginal bleeding 13 months earlier. Other pregnancy-related diseases were excluded following a negative serum human chorionic gonadotropin test, and the diagnosis was confirmed by Doppler ultrasonography. An emergency hysterectomy was subsequently performed due to the large amount of vaginal bleeding. Postoperative pathology confirmed the uterine arteriovenous malformation and demonstrated that the cause was a previous cornual pregnancy with placental implantation. The patient successfully recovered following the surgery.

A modified method for the culture of naturally HPV-infected high-grade cervical intraepithelial neoplasia keratinocytes from human neoplastic cervical biopsies.

Few studies on cervical intraepithelial neoplasia (CIN) keratinocyte cultures are available due to the numerous technical and methodological problems associated with the in vitro cultivation of these cells. The present study investigated an applicable and effective method for the in vitro cultivation of high-grade CIN keratinocytes from human neoplastic cervical biopsies. Human neoplastic cervical tissue sections were obtained and digested using type I collagen in order to dissociate the cells. The cells were seeded in tissue culture plastic plates that were coated with rat tail collagen type I and contained modified keratinocyte serum-free medium (K-SFM) supplemented with 5% fetal bovine serum. The medium was replaced with K-SFM on days 3, 5 and 7, respectively. The unattached cells were recovered and the cell viability was determined accurately using the Trypan Blue exclusion method. The expression of keratin 14 (K14), keratin 19 (K19), keratin 17 (K17) and P63 was assayed using immunofluorescence in order to identify the presence of CIN keratinocytes. The present results indicated that the attachment rate of CIN keratinocytes significantly increased between 56.75±1.76% on day 3 and 77.09±3.55% on day 5, and became relatively stable between days 5 and 7. The cell viability significantly decreased between 83.00±0.50% on day 5 and 68.17±1.04% on day 7. The passaged CIN keratinocytes maintained the original unequally sized, abnormally shaped morphology and did not undergo differentiation. In addition, the passaged CIN keratinocytes exhibited the same human papilloma virus (HPV) genotype that was detected in the original primary cells. K14 and K19 were expressed in the majority of the normal and CIN keratinocytes, whereas K17 and P63 were expressed only in high-grade CIN keratinocytes. The present study proposes a simple and practical method for rapidly obtaining highly purified naturally HPV-infected high-grade CIN keratinocytes from small neoplastic cervical tissues, and provides an appropriate first medium change time for the primary culture of CIN keratinocytes.

Evaluation of several screening approaches for detection of cervical lesions in rural Shandong, China.

PURPOSE: The study was designed to: (1) investigate the prevalence of high-risk human papillomavirus (HR- HPV) infection and cervical neoplasia; and (2) evaluate clinical performance of visual inspection with acetic acid/ Lugol's iodine (VIA /VILI), Pap smear, high-risk human papillomavirus (HR-HPV) DNA test for detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and (3) explore appropriate screening approach in rural areas of Shandong Province. MATERIALS AND METHODS: A total of 3,763 eligible women from Yiyuan County in Yimeng mountainous areas of rural Shandong, China, were enrolled and underwent Pap smear, HR-HPV DNA testing by Hybrid Capture 2 (HC2), and VIA /VILI tests. Women positive in any test were referred to colposcopy and biopsy as indicated. RESULTS: The prevalence of HR-HPV infection among all enrolled women was 11.1% and that in healthy women was 9.9%. In total 33 cases of CIN1, 16 cases of CIN2, 6 cases of CIN3 but none of cervical cancer were detected and the crude prevalence of CIN2+ was 0.58%. For detecting CIN2+, the sensitivity of HR-HPV DNA testing, VIA/VILI, Pap smear was 90.9%, 77.3%, 81.8%, respectively. Pap smear had the best specificity of 98.2%, followed by HR-HPV DNA testing with specificity of 89.4%, VIA/VILI had the lowest specificity of 81.2%. Colposcopy referral rate of HR-HPV DNA testing, VIA/VILI, Pap smear was 11.1%, 18.5%, 2.3%, respectively. CONCLUSIONS: Our results suggest that HR-HPV DNA testing alone might be appropriate for primary cervical cancer screening in rural low-resource areas of Shandong Province, China.

Efficacy of multiple low-dose photodynamic TMPYP4 therapy on cervical cancer tumour growth in nude mice.

OBJECTIVE: Photodynamic therapy (PDT) is an emerging therapeutic procedure suitable for the treatment of cervical cancer. However, the side effects of PDT are severe, including skin ulceration, so we designed an experiment to examine the effects of multiple low- dose photodynamic therapy of 5, 10, 15, 20-tetrakis(1- methylpyridinium-4-yl) porphyrin (Tmpyp4) on tumour growth by utilizing a model in nude mice implanted with Hela cervical cancer cells. MATERIALS AND METHODS: Female BALB/c nude mice (aged 5-6 weeks, weighing 18-20 g) were used. Hela cervical cancer cells were injected subcutaneously (1 x 10(7) cells/200 µL). Ten days after injection, the mice were divided into three groups (n=6), the A group of controls without any treatment, the B group receiving a single-treatment with Tmpyp4 (10 mg/kg, intratumor injection) and irradiation (blue laser, 108 J/cm(2)), and the C group given three-treatments with Tmpyp4 (10 mg/ kg, intratumor injection) and irradiation at intervals of two days. After starting treatment, tumours were measured every two days, to assess growth. At 2 weeks after the last treatment of C group, tumour tissue and organs were collected from each mouse to evaluate tumor histology and organ damage. RESULTS: Tumour growth in C group was significantly inhibited compared with A and B groups (P <0.05), without any injury to the skin and internal organs. CONCLUSION: Our novel findings demonstrated that multiple low-dose photodynamic therapy of Tmpyp4 could inhibit cervical cancer growth significantly with no apparent side effects.

[Needle-knife closed solution combined with minor adjusting of spine for the treatment of neck-shoulder syndrome].

OBJECTIVE: To evaluate therapeutic effect of the needle-knife closed solution combined with minor adjusting of spine for treatment of neck-shoulder syndrome. METHODS: From April 2010 to August 2011,120 patients with neck-shoulder syndrome were treated with the needle-knife closed solution combined with minor adjusting of spine, and included 45 males and 75 females and aged from 40 to 68 years old. The disease course was from 3 days to 10 years. After the operation, all patients taken the medicine of activating blood circulation herbs. At the 3rd, 7th, 10th day after operation, spinal rotation massage was performed on these patients. After the healing of the needle points, traditional Chinese medicine herb fumigation was applied on the needle points, and the patients were directed to do the cervical spine exercise. Therapeutic effect of the patients was evaluated by the neck disability index (NDI). RESULTS: All patients were followed up after 3 weeks' treatment. The pain of neck-shoulder was relieved, and the range of motion was improved,the NDI score lowered from 49.30 +/- 1.35 before treatment to 10.15 +/- 1.18 at 3 weeks after treatment (t = 2.116, P < 0.05). CONCLUSION: The needle-knife closed solution combined with minor adjusting of spine for the treatment of neck-shoulder syndrome can relieve the pain in the neck-shoulder and improved the motion of the neck. The key for the effect is accurate location before operation, sufficient adhesion solution during the operation and spinal minor adjusting after operation.

Establishment of a novel method for primary culture of normal human cervical keratinocytes.

BACKGROUND: Cervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro. METHODS: Normal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity. RESULTS: Our results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%. CONCLUSION: A novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Synergistic anticancer activity of 5-aminolevulinic acid photodynamic therapy in combination with low-dose cisplatin on Hela cells.

OBJECTIVE: Photodynamic therapy (PDT ) is a promising modality for the treatment of various tumors. In order to assist in optimizing treatment, we applied 5-ALA/PDT in combination with low-dose cisplatin to evaluate cytotoxicity in Hela cells. METHODS: Antiproliferative effects of 5-ALA/PDT and cisplatin, alone and in combination, were assessed using MTT assay. To examine levels of apoptosis, Hela cells treated with 5-ALA/PDT, and combination treatment were assessed with Annexin-V/PI by flow cytometry. To investigate the molecular mechanisms underlying alterations in cell proliferation and apoptosis, Western blot analysis was conducted to determine the expression of p53, p21, Bax and Bcl-2 proteins. RESULTS: MTT assays indicated that combination treatment obviously decreased the viability of Hela cells compared to individual drug treatment. In addition, it was confirmed that exposure of Hela cells to 5-ALA/PDT in combination with low-dose cisplatin resulted in more apoptosis in vitro. Synergistic anticancer activity was related to upregulation p53 expression and alteration in expression of p21, Bcl-2 and Bax. CONCLUSION: Our findings suggest that administration of 5-ALA/PDT in combination with the low-dose cisplatin may be an effective and feasible therapy for cervical cancer.

Significance of human telomerase RNA gene amplification detection for cervical cancer screening.

AIM: Liquid-based cytology is the most often used method for cervical cancer screening, but it is relatively insensitive and frequently gives equivocal results. Used as a complementary procedure, the high-risk human papillomavirus (HPV) DNA test is highly sensitive but not very specific. The human telomerase RNA gene (TERC) is the most often amplified oncogene that is observed in cervical precancerous lesions. We assessed genomic amplification of TERC in liquid-based cytological specimens to explore the optimal strategy of using this for cervical cancer screening. METHODS: Six hundred and seventy-one residual cytological specimens were obtained from outpatients aged 25 to 64 years. The specimens were evaluated by the Digene Hybrid Capture 2 (HC2) HPV DNA test and fluorescence in situ hybridization (FISH) with a chromosome probe to TERC (3q26). Colposcopic examination and histological evaluation were performed where indicated. RESULTS: The TERC positive rate was higher in the CIN2+ (CIN2, CIN3 and SCC) group than in the normal and CIN 1 groups (90.0% vs. 10.4%, p<0.01). In comparison with the HC2 HPV DNA test, the TERC amplification test had lower sensitivity but higher specificity (90.0% vs. 100.0%, 89.6% vs. 44.0%, respectively). TERC amplification test used in conjunction with the HC2 HPV DNA test showed a combination of 90.0% sensitivity and 92.2% specificity. CONCLUSION: The TERC amplification test can be used to diagnose cervical precancerous lesions. TERC and HPV DNA co-testing shows an optimal combination of sensitivity and specificity for cervical cancer screening.

Clinical and pathological factors related to the prognosis of chinese patients with stage Ib to IIb cervical cancer.

OBJECTIVE: The aim of this retrospective study is to analyze the clinical and pathological factors related to the prognosis of Chinese patients with stage Ib to IIb cervical cancer. METHODS AND RESULTS: 13 clinical pathological factors in 255 patients with stage Ib to IIb cervical cancer undergoing radical hysterectomy and systematic lymphadenectomy were analyzed to screen for factors related to prognosis. The cumulative 5-year survival of the 255 patients was 75.7%. The result of the univariate analysis suggested that clinical stage, cell differentiation, depth of cervical stromal invasion, parametrial tissue involvement, and lymph node metastasis were prognostic factors for patients with stage Ib to IIb cervical cancer (P<0.05). Compared with cases with involvement of iliac nodes, obturator nodes, or inguinal lymph nodes, cases with metastasis to the common iliac lymph nodes had a poorer prognosis (P<0.05). Cases with involvement of four or more lymph nodes had a poorer prognosis than those with involvement of three or fewer lymph nodes (P<0.05). Using multivariate Cox proportional hazards model regression analysis, non-squamous histological type, poor differentiation, parametrial tissue involvement, and outer 1/3 stromal invasion were found to be independently related to patients poor prognosis (P<0.05). CONCLUSION: Non-squamous histological type, poor cell differentiation, parametrial tissue involvement, and outer 1/3 stromal invasion are the independent poor prognostic factors for patients with stage Ib to IIb cervical cancer.

[Application of pathological diagnosis by rapid paraffin sections for biopsy in the diagnosis and treatment of cervical diseases].

OBJECTIVE: To evaluate the application of pathological diagnosis by rapid paraffin sections in the diagnosis and treatment of cervical diseases. METHODS: A total of 176 cases from our hospital between September 2009 and January 2010 with abnormal cervical cancer screening (including abnormal cytology result and high-risk HPV continuous positive) were randomly divided into 2 groups. Eighty-seven cases of them whose biopsy were got by Belinson forceps under the direction of colposcopy with rapid paraffin sections by ultrasonic histopathological rapid processor and BT transparent agents were selected as group A, while 89 cases with conventional paraffin sections were selected as group B. The production time and quality for paraffin sections were analyzed in the two groups. Those diagnosed as cervical intraepithelial neoplasia (CIN) II or even worse and some special patients with CINI in the two groups received surgery, including loop electrosurgical procedure (LEEP), cold knife conization (CKC), hysterectomy or radical hysterectomy. Tissue obtained after surgery was sent for routine pathological examination. If the results of postoperative routine pathological examination were inconsistent with the rapid or routine biopsy pathological examination, the heavier results were regard as the final diagnoses. The pathological results and diagnose accordance rates were recorded and compared between group A and group B. RESULTS: The quality of sections in two groups were all satisfied or basically satisfied to meet the diagnostic requirements. There were statistically significant difference in average production time between group A and B (40 minutes vs 24 hours, P<0.05). Thirty patients in group A and 32 patients in group B received surgery. The coincidence rate of biopsy pathological results and final diagnoses were 93% (28/30) for group A and 91% (29/32) for group B, in which there were not statistically significant difference (P>0.05). CONCLUSION: Rapid paraffin sections technology is safe, accurate and economical for rapid pathological diagnosis of cervical diseases, which is worthy for being widely used in hospitals.

Expressions of IGFBP-5, cFLIP in cervical intraepithelial neoplasia, cervical carcinoma and their clinical significances: a molecular pathology.

BACKGROUND: Insulin-like growth factor binding protein (IGFBPs) have been as potential tumor suppressors in the occurrence and development of tumors. Cellular Fas-associated death domain-like interleukin-1beta-converting enzyme (FLICE)-like inhibitory protein (cFLIP) contains a death effect domain (DED), which blocks death receptor pathway and inhibits apoptosis. METHODS: We collected normal cervical tissues from 28 subjects, CIN samples from 37 patients, and cervical cancer tissues from 40 patients. In these samples, we then measured the expression levels of IGFBP-5 and cFLIP via RT-PCR and immunohistochemistry, and we detected the presence of high-risk HPV by Hybrid capture II assays in cervical secretions provided by the subjects. RESULTS: significant differences in the expression of IGFBP-5 protein among the normal, CIN, and CC tissues (P < 0.05). The highest expression of IGFBP-5 protein was found in CIN stage II and III tissues, whereas the expression of IGFBP-5 in CC samples was decreased relative to controls. The expression level was affected by factors such as clinical stage, pathological differentiation, and lymph node metastasis. Relative to the controls, IGFBP-5 mRNA content was higher in the CC group and lower in the CIN group (P < 0.05). No expression of cFLIP protein or mRNA was detected in normal cervical tissues. However, the degree of pathological changes correlated with increasing expression of cFLIP protein and mRNA, and significant differences were therefore detected between groups (P < 0.05). The HPV infection rates in the CIN and CC groups were much higher than in the normal group (P < 0.05). CONCLUSION: IGFBP-5 expression is up-regulated in response to progression of CIN and down-regulated in invasive cervical carcinoma. Detection of IGFBP-5 and cFLIP expression levels, may prove particularly useful for diagnosing and differentiating CIN and CC.

[Surgical treatment of Lisfranc fracture-dislocations by reconstruction of three-column with Kirschner wire fixation].

OBJECTIVE: To explore the effect of reduction and fixation with multi-Kirschner wires for treatment of Lisfranc fracture-dislocations. METHODS: There were 49 patients (37 male and 12 female aged from 20 to 28 years old) involved in the study. According to the Myserson damage typing, type A in 12 cases, B1 in 3, B2 in 28, C1 in 4 and C2 in 2. Kirschner wires were applied to fix and reconstruct the three-column in three directions according to the structural characteristic of midfoot. Reconstruction of three-column needed not only to reduce and to fix every single column, but also to establish union of the columns. RESULTS: According to the evaluation of AOFAS for midfoot, 14 cases were excellent (90 to100), 22 cases good (80 to 89), 8 cases fair (70 to 79) and 5 cases poor (60 to 69), with an average score of 84.200+/-9.663. CONCLUSION: The diagnosis and treatment of Lisfranc fracture-dislocations should comply with the theory of three-column reconstruction of foot arch, which can achieve the static balance of biomechanics and provide a stable environment for healing of fracture and soft tissue. Reduction and fixation with multi-Kirschner wires is an effective treatment method for Lisfranc fracture-dislocations.

[Effect of extracellular signal regulated kinase signal pathway on apoptosis induced by MG262 in ovarian cancer cells].

OBJECTIVE: To investigate whether the proteasomes inhibitor MG262 exerts its anti-cancer function by inducing apoptosis in human ovarian cancer cells, and whether the extracellular signal regulated kinase (ERK) signaling pathway is involved in the regulation of apoptosis induction. METHOD: Human ovarian cancer cell line SKOV3 was incubated with different concentrations of MG262 for 24 and 48 hours. Cell viability was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at different time points of culturing. Flow cytometry was used to detect cell apoptosis rate. The expression of vascular endothelial growth factor (VEGF) was evaluated with western blot and enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression of phosphorylated ERK (p-ERK). RESULTS: The viability of SKOV3 cells was decreased by MG262 in a concentration-dependent fashion (P < 0.05). After 24 h incubation with MG262 at 1, 10, 20, 40, 60 and 80 nmol/L, the viability rates of SKOV3 were (94.6 +/- 3.1)%, (92.7 +/- 3.7)%, (89.5 +/- 7.7)%, (84.2 +/- 5.1)%, (82.0 +/- 7.4)% and (76.8 +/- 11.0)% respectively, and after 48 h incubation, those figures were further decreased to (91.3 +/- 10.1)%, (86.8 +/- 4.5)%, (74.6 +/- 4.2)%, (56.8 +/- 2.1)%, (49.3 +/- 4.5)% and (37.4 +/- 5.4)%, respectively (P < 0.05). Apoptosis rate of SKOV3 cells induced by MG262, PD98059 or their combination was (30.7 +/- 4.3)%, (26.8 +/- 8.6)% and (50.3 +/- 10.6)%, respectively, which were significantly different compared with controls (P < 0.05). In contrast to SKOV3 cells, apoptosis rate of 293T cells induced by MG262, PD98059 or their combination was (14.5 +/- 5.3)%, (16.2 +/- 7.5)% and (10.8 +/- 7.3)%, respectively, which were not significantly different compared with controls (P > 0.05). p-ERK expression decreased gradually in a time-dependent manner. And wild-type p53 expression was not significantly different. There was no significant difference between experimental and control 293T cells (P < 0.05). In addition, MG262 down-regulated VEGF secretion and expression in SKOV3 cells (P < 0.05). CONCLUSIONS: Proteasome inhibitors can induce apoptosis and inhibit cell proliferation and angiogenesis through ERK signal pathway in SKOV3 cells.

[Expression and significance of chemokine CXC receptor 3, 4 and their ligands at the early pregnancy decidua and villi].

OBJECTIVE: To explore the expression and significance of chemokine CXC receptor (CXCR) 3 and CXCR4 and their ligands (CXCL) at the early pregnancy decidua and villi. METHODS: Decidual mononuclear cells were isolated from the normal decidua of 5 - 8 weeks pregnant women by lymphocyte separation medium in vitro. CD(56)(+) natural killer (NK) cells were purified by dynabeads cell sorter kit. Purity and phenotype of CD(56)(+) decidua NK cells were analyzed by fluorescence-activated cell sorter (FACS). Gene expression of CXCR3 and CXCR4 in decidua NK cells and CXCL9, CXCL10 and CXCL12 in early pregnancy decidua and villi was assessed by RT-PCR. Protein expression of CXCL9, CXCL10 in normal endometrium and early pregnancy decidua was characterized and quantified by streptavidin-biotin peroxidase chain reaction (SP) immunohistochemistry and computered image analysis system. Correlations between the gray degree of CXCL9 and CXCL10 and the number of CD(56)(+)NK cells in upper tissue were analyzed by Spearman's correlation coefficient rank test. RESULTS: The phenotype of 98.7% decidua NK cells was CD(56)(bright). The genes of CXCR3 and CXCR4 were expressed in decidua NK cells and that of CXCL9 and CXCL10 were expressed in early pregnancy decidua and CXCL12 in early pregnancy villi. CXCL9 and CXCL10 were expressed in the cytoplasm of surface epithelia, glandular epithelia and stromal cells of early pregnancy decidua and were not expressed in villi by immunohistochemistry. The gray degree of CXCL9 and CXCL10 in the secretory phase endometrium (56 +/- 43, 59 +/- 47) was stronger than that in the proliferative phase (16 +/- 18, 8 +/- 14, P < 0.05) and reached the highest (143 +/- 35, 158 +/- 29, P < 0.05) in the early pregnancy decidua. The number of CD(56)(+) NK cell in the secretory phase endometrium (60 +/- 20) was more than that in the proliferative phase endometrium (23 +/- 4, P < 0.05) and was the most in the early pregnancy decidua (114 +/- 15, P < 0.05). The gray degree of CXCL9 in upper tissue had a positive correlation with the number of CD(56)(+) cells (r = 0.88, P < 0.05) and that of CXCL10 had a similar pattern to CXCL9 (r = 0.86, P < 0.05). CONCLUSION: The interactions between CXCL9, CXCL10 and CXCL12 expressed in decidua and villi and CXCR3, CXCR4 expressed in CD(56)(+) decidua NK cells may influence the CD(56)(+)NK cell recruitment at the maternal-fetal interface.

[Combination chemotherapy with etoposide and cisplatin for high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia].

OBJECTIVE: To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin (EP) regimen on the patients with high-risk, chemorefractory and recurrent gestational trophoblastic neoplasia (GTN). METHODS: Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively, 25 of 39 patients were of high-risk, 9 patients were chemorefractory and 5 patients were recurrent. All 39 patients were administrated with EP regimen, and 10 patients were assisted with surgery. All the patients were followed up. Clinical response, toxicity, the occurrence of secondary tumors of all patients, and the fertility of 30 patients whose fertility function was preserved were investigated. RESULTS: Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen. The average number of courses for each patient was 5.7. The total complete remission rate of the regimen was 74% (29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6. Nineteen patients achieved complete remission and 6 patients showed drug-resistant. The complete remission rate of the high-risk group was 76% (19/25). Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1. Six patients achieved complete remission and 3 patients showed drug-resistant again. The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4. Four patients achieved complete remission, 1 patient showed drug-resistance and died. Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen, but the bone marrow toxicity was slight and no grade IV side effect occurred. No fatal effect was found. Eight of 30 patients whose fertility fuction was preserved had become pregnant after recovery, with a total of 8 pregnancies. Among them, 2 were terminated by induced abortion, and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation. All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors. CONCLUSION: The EP regimen is effective and safe for the treatment of high-risk, chemorefractory and recurrent GTN.

[Clinical study of topotecan and cisplatin as first line chemotherapy in epithelial ovarian cancer].

OBJECTIVE: To evaluate efficacy and toxicity of topotecan and cisplatin (TP) as first line chemotherapy in epithelial ovarian cancer, and its effect on prognosis of the patients. METHODS: Totally 94 eligible patients with pathologically verified stage II - IV epithelial ovarian cancer were enrolled into 3 groups of this clinical trial. (1) TP group: 30 patients were treated with topotecan, 0.75 mg.m(-2).d(-1), for 5 days, and cisplatin, 75 mg/m(2), on day 1. (2) Paclitaxel and carboplatin (TC) group: 31 patients were treated with paclitaxel, 135 mg/m(2), on day 1, and carboplatin, given to an area under the curve (AUC) of 5, on day 1. (3) Cyclophosphamide and cisplatin (PC) group: 33 patients were treated with cyclophosphamide, 500 mg/m(2), on day 1, cisplatin 75 mg/m(2), on day 1. Cycles were repeated every 21 - 28 days. EFFICACY of the three combination regimens were evaluated after 6 - 8 courses. RESULTS: (1) EFFICACY: the overall response rate (ORR) in the TP group was 70%. Of the 30 patients, 8 achieved a complete response (CR) and 13 a partial response (PR). The ORR in the TC group was 77%. Of the 31 patients, 10 achieved a CR and 14 a PR. While the ORR in the PC group was 42%. Of the 33 patients, 5 achieved a CR and 9 a PR. There was no significant difference in clinical efficacy between TP group and TC group (P > 0.05). But there was a significant difference between TP group and PC group (P < 0.05). (2) Disease free survival (DFS): after median follow-up of 25 months, one-year disease free survival rate was 67% in TP group, 71% in TC group and 42% in PC group (P > 0.05). Two-year disease free survival rate was 57% in TP group, 64% in TC group and 39% in PC group (P > 0.05). (3) Overall survival (OS): One-year survival rate was 93% in TP group, 97% in TC group and 91% in PC group (P > 0.05). Two-year survival rate was 77% in TP group, 84% in TC group and 67% in PC group (P > 0.05). (4) TOXICITY: Grade III - IV myelosuppression was 60% (18/30) in TP group, 26% (8/31) in TC group and 30% (10/33) in PC group. The TP regimen had the greatest hematologic toxicity (P < 0.05). Nonhematologic toxicities were not significantly different among the three regimens (P > 0.05). CONCLUSIONS: As first line chemotherapy in epithelial ovarian cancer, TP regimen comparable to the standard chemotherapy regimen.

[Photodynamic effect of hematoporphyrin monomethyl ether on ovarian cancer cell line SKOV3].

BACKGROUND & OBJECTIVE: Photodynamic therapy (PDT) is a new treatment choice for ovarian carcinoma. Hematoporphyrin monomethyl ether (HMME) is a novel photosensitive reagent developed in China. This study was to investigate the photodynamic effect of HMME-based PDT on human epithelial ovarian cancer cell line SKOV3. METHODS: After an incubation with 30 microg/ml HMME for different time, the fluorescent image and intracellular location of HMME in SKOV3 cells were observed under a fluorescent microscope and laser scanning confocal microscope (LSCM). After being treated with different doses (5-50 microg/ml) of HMME and irradiated with different optical doses (1.5-12 J/cm(2)) of laser, the survival rate of SKOV3 cells was measured by MTT assay. Mechanisms of cell death during PDT was determined by Annexin V/PI double staining technique and analyzed by flow cytometry. RESULTS: Red fluorescence appeared shortly after administration of HMME and localized in cytoplasm; intracellular fluorescence intensity reached the peak after 3 h. High concentrations of HMME alone had cytotoxicity to SKOV3 cells, while laser irradiation alone had no effect on cell survival. Survival rate of SKOV3 cells was gradually decreased along with the increase of HMME concentration and laser dose, but such a trend diminished when HMME concentration reached 40 microg/ml. After treatment of HMME, the dead cells were predominantly necrosis cells. CONCLUSION: HMME has a photodynamic effect on SKOV3 cells.