Cancer genome scanning in plasma: detection of tumor-associated copy number aberrations, single-nucleotide variants, and tumoral heterogeneity by massively parallel sequencing.

BACKGROUND: Tumor-derived DNA can be found in the plasma of cancer patients. In this study, we explored the use of shotgun massively parallel sequencing (MPS) of plasma DNA from cancer patients to scan a cancer genome noninvasively. METHODS: Four hepatocellular carcinoma patients and a patient with synchronous breast and ovarian cancers were recruited. DNA was extracted from the tumor tissues, and the preoperative and postoperative plasma samples of these patients were analyzed with shotgun MPS. RESULTS: We achieved the genomewide profiling of copy number aberrations and point mutations in the plasma of the cancer patients. By detecting and quantifying the genomewide aggregated allelic loss and point mutations, we determined the fractional concentrations of tumor-derived DNA in plasma and correlated these values with tumor size and surgical treatment. We also demonstrated the potential utility of this approach for the analysis of complex oncologic scenarios by studying the patient with 2 synchronous cancers. Through the use of multiregional sequencing of tumoral tissues and shotgun sequencing of plasma DNA, we have shown that plasma DNA sequencing is a valuable approach for studying tumoral heterogeneity. CONCLUSIONS: Shotgun DNA sequencing of plasma is a potentially powerful tool for cancer detection, monitoring, and research.

Radiotherapy and chemoradiation after surgery for early cervical cancer.

BACKGROUND: This is an updated version of the original Cochrane review first published in Issue 4, 2009. There is an ongoing debate about the indications for, and value of, adjuvant pelvic radiotherapy after radical surgery in women with early cervical cancer. Certain combinations of pathological risk factors are thought to represent sufficient risk for recurrence, that they justify the use of postoperative pelvic radiotherapy, though this has never been shown to improve overall survival, and use of more than one type of treatment (surgery and radiotherapy) increases the risks of side effects and complications. OBJECTIVES: To evaluate the effectiveness and safety of adjuvant therapies (radiotherapy, chemotherapy followed by radiotherapy, chemoradiation) after radical hysterectomy for early-stage cervical cancer (FIGO stages IB1, IB2 or IIA). SEARCH METHODS: For the original review, we searched the Cochrane Central Register of Controlled Trials (CENTRAL), Issue 4, 2008. The Cochrane Gynaecological Cancer Group Trials Register, MEDLINE (January 1950 to November 2008), EMBASE (1950 to November 2008). We also searched registers of clinical trials, abstracts of scientific meetings, reference lists of included studies and contacted experts in the field. For this update, we extended the database searches to September 2011 and searched the MetaRegister for ongoing trials. SELECTION CRITERIA: Randomised controlled trials (RCTs) that compared adjuvant therapies (radiotherapy, chemotherapy followed by radiotherapy, or chemoradiation) with no radiotherapy or chemoradiation, in women with a confirmed histological diagnosis of early cervical cancer who had undergone radical hysterectomy and dissection of the pelvic lymph nodes. DATA COLLECTION AND ANALYSIS: Two review authors independently abstracted data and assessed risk of bias. Information on grade 3 and 4 adverse events was collected from the trials. Results were pooled using random-effects meta-analyses. MAIN RESULTS: Two RCTs, which compared adjuvant radiotherapy with no adjuvant radiotherapy, met the inclusion criteria; they randomised and assessed 397 women with stage IB cervical cancer. Meta-analysis of these two RCTs indicated no significant difference in survival at 5 years between women who received radiation and those who received no further treatment (risk ratio (RR) = 0.8; 95% confidence interval (CI) 0.3 to 2.4). However, women who received radiation had a significantly lower risk of disease progression at 5 years (RR 0.6; 95% CI 0.4 to 0.9).Although the risk of serious adverse events was consistently higher if women received radiotherapy rather than no further treatment, these increased risks were not statistically significant, probably because the rate of adverse events was low. AUTHORS' CONCLUSIONS: We found evidence, of moderate quality, that radiation decreases the risk of disease progression compared with no further treatment, but little evidence that it might improve overall survival, in stage IB cervical cancer. The evidence on serious adverse events was equivocal.

Adjuvant radiotherapy and chemoradiation after surgery for cervical cancer.

BACKGROUND: There is an ongoing debate about the indications for, and value of, adjuvant pelvic radiotherapy after radical surgery in women with early cervical cancer. Certain combinations of pathologic risk factors are thought to represent sufficient risk for recurrence, that they justify the use of post-operative pelvic radiotherapy, though this has never been shown to improve overall survival, and use of more than one type of treatment (surgery and radiotherapy) increases the risks of side-effects and complications. OBJECTIVES: To evaluate the effectiveness and safety of adjuvant therapies (radiotherapy, chemotherapy followed by radiotherapy, chemoradiation) after radical hysterectomy for early stage cervical cancer (FIGO stages IB1, IB2 or IIA). SEARCH STRATEGY: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), Issue 4, 2008. The Cochrane Gynaecological Cancer Group Trials Register, MEDLINE (January 1950 to November 2008), EMBASE (1950 to November 2008). We also searched registers of clinical trials, abstracts of scientific meetings, reference lists of included studies and contacted experts in the field. SELECTION CRITERIA: Randomised controlled trials (RCTs) that compared adjuvant therapies (radiotherapy, chemotherapy followed by radiotherapy, or chemoradiation) with no radiotherapy or chemoradiation, in women with a confirmed histological diagnosis of early cervical cancer who had undergone radical hysterectomy and dissection of the pelvic lymph nodes. DATA COLLECTION AND ANALYSIS: Two review authors independently abstracted data and assessed risk of bias. Information on grade three and four adverse events was collected from the trials. Results were pooled using random effects meta-analyses. MAIN RESULTS: Two RCTs, which compared adjuvant radiotherapy with no adjuvant radiotherapy, met the inclusion criteria; they randomised and assessed 397 women. Meta-analysis of these two RCTs indicated no significant difference in survival at five years between women who received radiation and those who received no further treatment (Relative risk (RR) = 0.8, 95% Confidence interval (CI): 0.3 to 2.4). However, women who received radiation had a significantly lower risk of disease progression at five years (RR = 0.6, 95% CI 0.4 to 0.9).Although the risk of serious adverse events was consistently higher if women received radiotherapy rather than no further treatment, these increased risks were not statistically significant, probably because the rate of adverse events was low. AUTHORS' CONCLUSIONS: We found evidence, of moderate quality, that radiation decreases the risk of disease progression compared with no further treatment, but little evidence that it might improve overall survival. The evidence on serious adverse events was equivocal.

Analysis of human papillomavirus type 18 load and integration status from low-grade cervical lesion to invasive cervical cancer.

The clinical value of viral load and integration testing for human papillomavirus (HPV) remains unclear. Data on HPV type 18 (HPV18) is limited. We examined the HPV18 viral load and integration status of 78 women with normal cervix or neoplasia. While the crude viral load appeared to increase with lesion severity, the association was not significant after normalization with sample cellularity. Unlike reports for HPV16, the amino-terminal 1 region of HPV18 E2 was most frequently (71.0%) disrupted, representing the best marker for integration. A substantial proportion (57.1%) of invasive cancers harbored only the episomal genome, thus jeopardizing the clinical value of integration testing. A large proportion (41.7%) of normal/low-grade lesions showed viral integration, suggesting that integration of HPV18 starts early and is unlikely to be a sole determinant for progression. Interpretation of viral load should take into account the form of HPV infection as single infections had significantly higher viral loads than coinfections (P = 0.046). More data generated from routinely collected samples are warranted to verify the clinical value of viral load and integration testing. Viral load quantitation for HPV18 is premature for clinical use at this stage.

Profile of viral load, integration, and E2 gene disruption of HPV58 in normal cervix and cervical neoplasia.

The clinical utility of viral-load and integration status of human papillomavirus (HPV) infection remains uncertain. We examined 75 women infected with HPV58, a worldwide rare type found to be prevalent in cervical cancers in eastern Asia. Viral load was significantly higher for cervical intraepithelial neoplasia (CIN) 1/2, but those for a normal control group and for CIN 3 or cancer overlapped substantially. A pure integrated genome was found for all lesion grades, giving a poor positive predictive value (23.1%) for cancer. The pure episomal form's negative predictive value for cancer was only 76.3%. Mixed patterns of E2 gene disruption were common and often involved the amino-terminal and hinge regions. Disruption of the whole E2 gene was rare and was restricted to high-grade lesions. The HPV58 variant E67-HK-2 was more likely to exist in the pure episomal form. Routinely collected cervical samples contain a heterogeneous population of viruses, hampering the application of viral load and integration testing in clinical settings.

Association between HLA-DRB1 polymorphism, high-risk HPV infection and cervical neoplasia in southern Chinese.

Multiple determinants are involved in the progression of human papillomavirus (HPV)-infected cervical lesion to invasive cancer. Human leukocyte antigen (HLA) polymorphism seems to play a role. This study examined the association between HLA-DRB1 polymorphism, high-risk HPV infection, and the development of cervical neoplasia in southern Chinese. Three hundred and seventy women with cervical neoplasia (43 cervical intraepithelial neoplasia grade II, 154 grade III, and 173 invasive cancers) and 323 controls were recruited for HLA-DRB1 typing by a sequence-based approach. Cervical specimens were collected for HPV detection by a consensus primer-based polymerase chain reaction, and with the type of HPV identified by hybridization with type-specific oligonucleotide probes. A protective effect of HLA-DRB1*12 for cervical neoplasia was observed, and with stronger associations when subgroup analyses were carried out for patients infected with HPV16 and HPV58. The protective effect of HLA-DRB1*13 that had been reported from other populations was not observed. The data obtained in this study showed that HLA-DRB1*03 conferred a higher risk for HPV18-infected, but not for HPV16-, HPV52-, or HPV58-infected cervical lesions. Although, HPV52 was reported as uncommon worldwide, it was found to be the second most prevalent type in the southern Chinese population. However, no additional risk association was observed when subgroup analyses were performed for HPV52-infected patients. The current study shows that, among southern Chinese, the outcome of HPV-infected cervical lesions is associated with HLA-DRB1 polymorphism. These associations often vary with the type of HPV infection.

HLA-DQB1 polymorphisms and risk for cervical cancer: a case-control study in a southern Chinese population.

OBJECTIVES AND METHODS: HLA II DQB1 polymorphisms have been shown to associate with cervical cancer risk, but results varied among different populations. In this study, the HLA DQB1 alleles among 221 southern Chinese women with cervical intraepithelial neoplasia grade III (CIN III)/invasive cervical carcinoma (ICC) were compared to 191 controls. RESULTS: The frequency of DQB1*03 was significantly lower among ICC overall as compared to controls (65.4% vs. 79.1%, odds ratio [95% confidence interval]: 0.50 [0.28-0.88], corrected p-value: 0.04). The protective association of DQB1*03 remained significant for human papillomavirus (HPV) 16-positive ICC, but not for HPV16-negative cases. This is in contrast to studies on European populations where DQB1*03 was associated with an increased risk for ICC. In the current study, 70.1% of the HPV16 isolates were Asian variants, and 28.0% were European variants. However, no significant association between HPV16 variant and DQB1*03 distribution was observed. HPV52 and HPV58 were found respectively in 16.3% and 10.0% of CIN III/ICC, which were higher compared to that of Europe and North America. Further analyses revealed a positive risk association between DQB1*06 and HPV58-positive CIN III/ICC (3.68 [1.37-9.92], corrected p-value: 0.012). CONCLUSION: The host genetics and the distribution of HPV types/variants may account for the observed differences among southern Chinese and other populations.

Lack of site-specific production of decidual alpha-2 macroglobulin in human pregnancy.

BACKGROUND: Alpha-2 Macroglobulin (A2M) is a protease inhibitor that is present in both human and rat decidual tissue. In mice, decidual A2M prevents excessive trophoblastic invasion; however, its role in human decidual tissue is unknown. It is possible that A2M may also influence trophoblast invasion in human pregnancy, which would be reflected in increased A2M production in decidua basalis. The aim of the current study was to determine and compare A2M production from first trimester human decidua basalis and decidua parietalis. METHODS: Human decidual tissues were obtained from patients undergoing surgical termination at 9 to 12 gestational weeks. Strips of decidua basalis and decidua parietalis were obtained by uterine curettage under real-time ultrasound guidance. Tissue samples were fixed in 10% formalin or snap-frozen for immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, respectively. Protein and mRNA production between the two sites were compared using the Mann-Whitney U test. RESULTS: Paired basal and parietal decidua were analyzed by immunohistochemistry (n = 9) and by RT-PCR (n = 10). There was no significant difference in A2M mRNA expression between decidua basalis and decidua parietalis (P = .5). Immunohistochemical staining intensity for A2M protein was significantly higher in basalis than in parietalis (P = .004), but the extent of positively stained cells were not significantly different (P = .051). Strong A2M staining in decidua basalis was mainly localized in the intracellular storage vesicles, which may suggest a role of A2M in this site. CONCLUSIONS: We conclude that the expression pattern of A2M in human decidua basalis and decidua parietalis is not consistent with an important role of this gene during the observed gestational period. Contrary to its role in rodent implantation, A2M is probably not involved in regulating human implantation and trophoblastic invasion during this gestational window frame.

Placental transfer of ondansetron during early human pregnancy.

BACKGROUND AND OBJECTIVE: Nausea and vomiting are common conditions that occur during early pregnancy and can be disabling. Ondansetron had been used in pregnant women when treatment with conventional antiemetics has failed; however, the safety and tolerability of this relatively new antiemetic drug during pregnancy is still uncertain. The objective of this study was to quantify the placental transfer of ondansetron in the first trimester of human pregnancy. PATIENTS AND METHODS: This was a prospective, observational study. Forty-one patients who requested surgical termination of pregnancy at the first trimester were administered three doses of ondansetron 8 mg before surgery. Maternal venous blood, coelomic fluid, amniotic fluid and fetal tissue were collected from each patient for analysis of ondansetron by liquid chromatography-mass spectometry. RESULTS: Ondansetron was found in all samples. Drug concentration in fetal tissue was significantly higher than that in the amniotic fluid and similar to that in the coelomic fluid. The median (interquartile range) fetal/maternal ratio was 0.41 (0.31-0.52) and there were no significant correlations between ondansetron concentrations in each compartment and gestational age. CONCLUSIONS: A significant amount of ondansetron was present in all embryonic compartments. The developmental significance of this drug exposure requires further investigation, i.e. whole embryo culture.

Is common iliac lymph node dissection necessary in early stage cervical carcinoma?

OBJECTIVES: Nodal metastasis is one of the most important prognostic factors in early stage cervical carcinoma and has an immense impact on the subsequent management. Thus, searching for nodal metastasis by pelvic lymphadenectomy is an integral part in the surgical management of cervical carcinoma. Complete nodal clearance of lymphatic tissue up to 2 cm above the bifurcation of common iliac vessels is therefore performed as a routine in our unit. The aim of this study is to investigate the incidence and pattern of pelvic lymph node metastases in patients with early stage cervical carcinoma to determine the role of common iliac node dissection in the surgery. METHODS: We retrospectively reviewed 174 operation and histopathology reports of patients who underwent pelvic lymphadenectomy because of stage IA2 to IIA cervical carcinoma. Lymph nodes collected below and above the bifurcation of common iliac vessels were labeled as pelvic nodes and common iliac nodes, respectively. The incidence and distribution of nodal metastases were analyzed. RESULTS: Complete and selective pelvic lymphadenectomy was performed in 163 and 11 patients, respectively. Nodal metastasis was documented in 35 (20.1%) patients. Pelvic and common iliac nodes were involved in 34 and 8 cases, respectively. All except one patient with common iliac node metastases were also found to have pelvic node metastasis. CONCLUSIONS: In early stage cervical carcinoma, isolated common iliac lymph node metastasis is rare, especially in cases without associated high risk factors. Less extensive pelvic lymphadenectomy may be considered in these patients in order to reduce operation morbidity and time.

HLA-B alleles, high-risk HPV infection and risk for cervical neoplasia in southern Chinese women.

A population-based study was conducted on 256 southern Chinese with cervical intraepithelial neoplasia grade III (CIN III) or invasive cervical cancer (ICC) and on 258 controls to examine the associations between HLA-B alleles, infection with high-risk human papillomaviruses (HPVs) and the development of cervical neoplasia. HLA-B15 was found to be protective for CIN III/ICC overall (p(corrected) = 0.003), and for HPV52-positive CIN III/ICC (p(corrected) = 0.003). A marginal protective effect of B15 was observed for HPV16-positive CIN III/ICC, but no significant associations were revealed for HPV18- or HPV58-positive cases. None of the HLA-B alleles were found to confer an increased risk for cervical neoplasia. HLA-B15 is common among Asian for whom HPV52, a worldwide uncommon HPV type, also exists in a relatively high prevalence. It would also be worthwhile to assess the association between HLA-B15, HPV52 and cervical cancer in other Asian populations.

Sexual dysfunction after a first trimester induced abortion in a Chinese population.

OBJECTIVE: To evaluate the short-term effect of a first trimester induced abortion on sexuality in a Chinese population. STUDY DESIGN: This is a cross-sectional study. One hundred and four women attending the post-abortion clinic 3-4 months after a first trimester induced abortion were recruited. They completed a self-administered questionnaire anonymously. The associations of various factors with sexual behavior after abortion were analyzed by the Chi-square test. RESULTS: More than 30% of the women reported a reduction in both frequency of vaginal intercourse and sexual desire. Their partners also had a reduction in sexual desire. The reduction of vaginal intercourse is more significant among young, unmarried women and those not having sterilization (P=0.01). In addition, the prevalence of reduction in sexual desire and enjoyment among those women who had repeated abortion was significantly higher (P<0.01). CONCLUSION: The sexuality of both women and their partners are affected after abortion. It may be due to both psychological trauma and the worry of another unwanted pregnancy. Proper counseling may relieve their anxiety and improve sexual lives.

Placental transfer of Zidovudine in first trimester of pregnancy.

OBJECTIVE: Zidovudine is one of the most common antiretroviral drugs used to prevent vertical transmission of human immunodeficiency virus. However, it is not recommended for use in the first trimester of pregnancy because of reservations about its potential teratogenicity during the organogenesis phase. The objective of this study was to investigate the placental transfer of zidovudine in the first trimester of human pregnancy. METHODS: Twenty-six pregnant women were given 2 oral doses of zidovudine (200 mg) before first trimester surgical termination of pregnancy. Maternal blood, fetal tissue, and coelomic and amniotic fluid were collected for drug analysis. RESULTS: Zidovudine was detected in all samples of maternal serum and fetal tissue but present in only 7 samples of amniotic and coelomic fluid. Zidovudine concentration in fetal tissue was similar to that of maternal serum. The median fetal/maternal ratio was 0.92 and was not associated with gestational age (r = 0.03, P = .89). CONCLUSION: Zidovudine crossed the first trimester human placenta readily and achieved the level of maternal serum rapidly. Patients who choose to take zidovudine in first trimester of pregnancy should be counseled about the potential fetal effects.

Human decidual production of hepatocyte-growth factor is not influenced by trophoblastic invasion in vivo.

OBJECTIVE: To evaluate and compare production of hepatocyte growth factor (HGF) from human first-trimester implantation-site decidua (decidua basalis) and nonimplantation site decidua (decidua parietalis), and hence to determine whether human trophoblast invasion in vivo is associated with increased decidual HGF production. DESIGN: Controlled prospective study. SETTING: University hospital-based study. PATIENT(S): Ten women undergoing first-trimester termination of singleton pregnancy for psychosocial reasons without preexisting medical or gynecologic diseases. INTERVENTION(S): Decidual samples surgically excised and processed for paraffin-embedded immunohistochemistry and for reverse transcription-polymerase chain reaction (RT-PCR) studies. MAIN OUTCOME MEASURE(S): Protein and mRNA production in decidua basalis and decidua parietalis by immunohistochemistry and RT-PCR, respectively. RESULT(S): No statistically significant difference was found between decidua basalis and decidua parietalis in HGF protein or mRNA production. Immunohistochemical analysis (n = 9) showed a mean score of 3.28 +/- 2.37 for decidua basalis and 3.61 +/- 2.66 for decidua parietalis. Semiquantitative analysis of HGF mRNA expression between the two sites showed no statistically significant difference (n = 10) CONCLUSION(S): Human decidual production of HGF is not influenced by trophoblastic invasion in vivo.

Nodal detection in malignant melanoma of the vagina using laparoscopic ultrasonography.

BACKGROUND: Primary malignant melanoma of the vagina is a rare variant of melanoma. It has worse prognosis compared to nongenital melanomas or other vaginal malignant neoplasms. CASE: A 40-year-old Chinese was diagnosed vaginal melanoma. Laparoscopic ultrasonography (USG) was used to search for abnormal pelvic and abdominal lymph nodes. Two metastatic pelvic lymph nodes were detected and excised. The vaginal tumour was removed by hysterectomy and partial vaginectomy. Despite a clear surgical margin and adjuvant radiotherapy, the patient died shortly after the operation. CONCLUSION: Patient with vaginal melanoma has grave prognosis, especially when metastatic disease presents. Radical surgery appears unjustified as a routine, it is essential to exclude lymphatic and distant metastases before embark to radical surgery. This report presents the first case of laparoscopic ultrasonographic detection of metastatic pelvic lymph nodes in patient with vaginal melanoma.