[Application of 21-gene recurrence risk score in patients with breast cancer].

Objective: To investigate the relationship between 21-gene recurrence risk score (21-Gene RS) and the prognosis and clinicopathological features of hormone receptor (HR) positive, HER2-negative early breast cancer patients who did not receive neoadjuvant therapy. Methods: A total of 469 patients with HR positive and HER2-negative early breast cancer who received surgical treatment in the First Affiliated Hospital, Zhejiang University School of Medicine from January 2014 to October 2017 were selected. Their clinicopathological data were retrospectively analyzed. Tumor tissue samples were collected from patients, and the expression of 21-gene was detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). The 21-Gene RS was calculated according to the Trial Assigning Individualized Options for Treatment (TAILORx) RS grouping and National Surgical Adjuvant Breast and Bowel Project B-20 (NSABP B-20) RS grouping principles. Patients were divided into low (21-Gene RS<11 or 21-Gene RS<18), intermediate (11≤21-Gene RS<26 or 18≤21-Gene RS<31) and high (21-Gene RS≥26 or 21-Gene RS≥31) risk groups, and the clinicopathological features and prognostic differences of patients in different risk groups were compared. Statistical data were compared by chi-square test. Survival analysis was performed using Kaplan-Meier curve analysis and the differences between groups were compared using Log-rank test. Multivariate analysis was conducted by COX regression analysis. Results: Based on TAILORx RS grouping, the proportions of low-risk, intermediate-risk and high-risk groups among the 469 patients were 18.8% (88/469), 48.2% (226/469) and 33.0% (155/469), respectively. Based on NSABP B-20 RS grouping, the proportion of low-risk, intermediate-risk and high-risk groups were 43.1% (202/469), 37.5% (176/469) and 19.4% (91/469), respectively. The association of 21-Gene RS with histological grading, luminal typing, Ki-67 expression, and chemotherapy and treatment modalities were statistically significant (P<0.05) regardless of TAILORx RS grouping or NSABP B-20 RS grouping. Kaplan-Meier survival curve suggested poor prognosis in high-risk group (P<0.05, Log-rank test). Multivariate COX regression analysis showed that surgical method and 21-Gene RS were risk factors affecting the prognosis of patients. Conclusions: 21-Gene RS is significantly associated with the prognosis of patients with HR-positive, HER2-negative, early-stage breast cancer not receiving neoadjuvant therapy, as well as with their clinicopathological characteristics such as patients' histologic grade, luminal typing, Ki-67 expression, and whether or not they are treated with chemotherapy or other treatment modalities.The 21-Gene RS threshold of 11 and 26 or 18 and 31 can be used to grade the prognosis in Chinese patients with early-stage breast cancer. More researches are needed to guide the selection of postoperative adjuvant therapy for patients with HR-positive and HER2-negative early-stage breast cancer.

[Performance of point-of-care testing for cervical cancer screening].

Objective: To evaluate the performance of point-of-care testing for cervical cancer and precancerous lesions screening. Methods: In September 2020, 197 and 273 women were selected by using simple random sampling method from "self-sampling" cohort and "physician-sampling" cohort established in Xiangyuan county, Shanxi Province, China, respectively. Cervical exfoliated cells were collected by women themselves or gynecologists. All samples were detected by POCT and women with positive result were directly referred for colposcopy. Subsequently, all the samples were detected by careHPV and PCR test. Colposcopy and punch biopsy were performed for women with POCT negative but careHPV or PCR test positive at another visit. Using histopathological diagnosis as the gold standard, we calculated sensitivity, specificity and drew the receiver operating characteristic (ROC) curves. The accuracy of POCT was analyzed and compared to that of careHPV and conventional PCR test in cervical cancer and precancerous lesions screening. Results: The median (Q1 , Q3) age of 470 women was 51 (45, 57) years old. Based on self-sampling, the sensitivity and specificity of POCT for CIN2+ were 100.00% (95%CI: 56.56%-100.00%) and 28.95% (95%CI: 22.97%-35.76%), respectively. Compared with POCT, POCT HPV16/18 test had similar sensitivity and higher specificity of 89.47% (95%CI: 84.30%-93.08%). Self-sampling POCT HPV16/18 test had an AUC of 0.947 (95%CI:0.910-0.985), which was higher than that of careHPV and PCR test. Physician-sampling POCT test had 100.00% sensitivity (95%CI: 64.57%-100.00%) and 55.85% specificity (95%CI: 49.83%-61.70%) for detecting CIN2+. POCT HPV16/18 test had lower sensitivity (71.43%, 95%CI: 35.90%-91.76%) and higher specificity (92.45%, 95%CI: 88.63%-95.06%). POCT HPV16/18 test generally showed similar AUC on both self-collected samples and clinician-collected samples (0.947 vs 0.819, P=0.217). Conclusion: POCT HPV16/18 test is an effective method with relatively high sensitivity and specificity for cervical cancer screening.

[Impact of previous cesarean delivery on pregnancy outcomes of in vitro fertilization and frozen-thawed embryo transfer].

Objective: To investigate the impact of a previous cesarean delivery on pregnancy outcomes of in vitro fertilization and frozen-thawed embryo transfer (FET). Methods: The clinical data of 1 179 patients who received in vitro fertilization and FET in Tianjin Central Hospital of Gynecology Obstetrics from January 2014 to May 2019 and had a history of the previous delivery were retrospectively analyzed. The patients were divided into four groups according to different previous delivery history and the number of embryo transferred: group A (single embryo transfer group with cesarean delivery history, n=338), group B (single embryo transfer group with vaginal delivery history, n=78), group C (double embryo transfer group with cesarean delivery history, n=444), and group D (double embryo transfer group with vaginal delivery history, n=319). The 1∶1 propensity score based on age, body mass index (BMI), infertility duration, basal FSH, basal LH, number of oocytes retrieved and high-quality embryo rate was used to match group A and B (caliper value=0.15), group C and D (caliper value=0.05), and group A and C (caliper value=0.01) respectively to reduce the influence of selection bias. The clinical pregnancy outcomes of patients were compared. Results: (1) Group A and group B were single embryo transfer groups with a total of 77 pairs of matched patients. There were no statistically significant differences in clinical pregnancy rate [42.9% (33/77) vs 45.5% (35/77)], miscarriage rate, preterm birth rate, and neonatal birth weight (all P>0.05). (2) Group C and group D were double embryo transfer groups with a total of 304 pairs of matched patients. The clinical pregnancy rate [42.4% (129/304)] and twin pregnancy rate [9.5% (29/304)] of Group C were significantly lower than those of Group D [53.0% (161/304), 15.5% (47/304) respectively; both P<0.05). There were no statistically significant in miscarriage rate, preterm birth rate and neonatal birth weight between the two groups (all P>0.05). (3) Groups A and C matched 318 pairs of patients. The two groups had no statistical significances in clinical pregnancy rate [38.4% (122/318) vs 45.6% (145/318)], miscarriage rate and preterm birth rate (all P>0.05), but the twin pregnancy rate in group C was significantly higher than that of group A [11.3% (36/318) vs 0.3% (1/318), P<0.01). (4) The occurrence of the low-birth-weight infant were related to gestational age (OR=0.41, 95%CI: 0.32-0.54) and twin pregnancy (OR=4.44, 95%CI: 1.93-10.21), and the occurrence of macrosomia was related to BMI (OR=1.18, 95%CI: 1.06-1.32). Moreover, the previous delivery method was not related to the neonatal birth weight (P>0.05). Conclusions: Patients with different delivery histories receive FET therapy, the pregnancy outcomes of single embryo transfer are not significantly different, and the success rate of double embryo transfer in patients with a cesarean delivery history is low. The neonatal birth weight is not related to the history of the cesarean section. It is recommended that patients with a cesarean delivery history choose elective single embryo transfer to ensure the success rate and to reduce the risk.

[Effect of body mass index on the assisted reproductive outcome of frozen-thawed embryo transfer in patients with polycystic ovary syndrome].

Objective: To investigate the effect of body mass index (BMI) on clinical pregnancy and neonatal outcomes in patients with polycystic ovary syndrome (PCOS) during frozen-thawed embryo transfer. Methods: A total of 650 patients with PCOS who received routine in vitro fertilization or intracytoplasmic sperm injection treatment for frozen-thawed embryo transfer from June 2014 to June 2019 in Tianjin Central Hospital of Gynecology Obstetrics were retrospectively analyzed. According to BMI, PCOS patients were divided into group A (18.5≤BMI<23 kg/m2, n=253), group B (23≤BMI<25 kg/m2, n=167), and group C (BMI≥25 kg/m2, n=230). The general information, clinical pregnancy outcomes, pregnancy complications, the incidence of macrosomia and low-birth-weight infants were compared in the three groups, and the influencing factors of neonatal birth weight were analyzed. Results: The embryo implantation rate, clinical pregnancy rate, and ongoing pregnancy rate all showed downward trend with the increase of BMI, but the differences were not statistically significant (all P>0.05). The live birth rate in group C [47.0% (108/230)] was significantly lower than those in groups A and B, with statistical significance (χ²=7.43, P=0.024). The late miscarriage rate was higher in group C [9.4% (13/139)] than in groups A and B (χ²=7.66, P=0.022). The birth rates of macrosomia in groups B [22.2% (16/72)] and group C [21.1% (16/76)] were significantly higher than that in group A, and the difference was statistically significant (χ²=14.15, P=0.001). There was no statistically significant difference in the incidence of gestational diabetes between the three groups (χ²=3.81, P=0.149). The incidence of hypertension disorders complicating pregnancy increased with the increase of BMI, and the difference was not statistically significant (P>0.05). Regression analysis showed that macrosomia was significantly associated with maternal pre-pregnancy BMI and gestational weeks, and the risk of macrosomia increased by 15% (95%CI: 3%-28%) for every increase in maternal BMI. Conclusions: The embryo implantation rate, clinical pregnancy rate, and ongoing pregnancy rate of PCOS patients in frozen-thawed embryo transfer cycles show downward trend with the increase of BMI. Obese patients with PCOS have a significant increase in late miscarriage rate and a significant decrease in live birth rate. The incidence of hypertension disorders complicating pregnancy in PCOS patients in the obese group has an increasing trend, and the birth rate of macrosomia has increased significantly. Therefore, it is recommended that obese women with PCOS lose weight scientifically before pregnancy to improve pregnancy and neonatal outcomes.

[Effect of the number of previous spontaneous abortions on the first in vitro fertilization cycle].

Objective: To investigate the effect of the number of previous spontaneous abortions on the first in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) cycle. Methods: A retrospective case-control study was conducted to analyze the clinical data of 1 279 patients who received IVF/ICSI treatment for the first time from July 2014 to July 2018 in Tianjin Central Hospital of Gynecology Obstetrics;they were divided into 0 time group (group A, n=924), 1 time group (group B, n=267) and 2 times group (group C, n=88) for comparison, according to the previous frequency of spontaneous abortions. Results: There were no statistically significant differences in age, basal testosterone, estradiol, progesterone, prolactin and embryo quality in group A, B and C (all P>0.05). The biochemical pregnancy rate of group C (9.1%) was higher than those of the other two groups (4.1% and 4.1%; all P>0.05). The clinical pregnancy rate of group A (42.5%) [>group B (40.4%) and >group C (35.2%)] was not statistically significant(P>0.05).Early abortion rate in group A (8.9%) was0.05). Conclusions: Women with a history of one- or two-time spontaneous abortion have no obvious effect on embryo quality, and have a negative impact of clinical pregnancy rate, early abortion rate, live birth rate in the first IVF/ICSI cycle; especially for patients with a history of two times spontaneous abortion, early abortion rate has a significant increase, live birth rate decreases significantly, suggesting that patients with a history of two times of spontaneous abortion should find the causes of miscarriage according to the process of recurrent spontaneous miscarriage and receive treatment if necessary before subsequent pregnancy.

[Study on the distribution and related factors of birth weight of live births in Guangxi Zhuang Autonomous Region of China from 2016 to 2018].

Objective: To investigate the distribution and related factors of birth weight of live births and full-term infants in Guangxi Zhuang Autonomous Region of China. Methods: Based on Guangxi women and children information system from 2016 to 2018, a large real-time database about maternal and live-birth information was established. It covered 1 712 midwifery institutions in Guangxi. A total of 2 394 240 cases of live births were collected and 2 243 129 cases of which were full-term infants. The multivariate logistic regression model was used to analyze the related factors of low birth weight. Results: The birth weight of 2 394 240 live births, (3 123.49±461.08) g, in Guangxi was approximately normal distribution with a peak distribution to the left. The incidence of low birth weight was 8.05%, and the incidence of macrosomia was 2.07%. The incidence of low birth weight was 10.92% for the puerpera with body mass index (BMI, kg/m(2)) <18.5, 16.82% for the puerpera with height <145 cm, 8.92% for the puerpera with age <20 years old, 7.67% for the puerpera with age ≥35 years old, and 54.65% for the puerpera with premature birth. The birth weight of 2 243 129 full-term infants, (3 176.01±400.78) g, was approximately normal distribution with a peak distribution to the right. The incidence of low birth weight was 2.97%, and the incidence of macrosomia was 2.19%. The incidence of low birth weight was 4.73% for puerpera with BMI<18.5, 8.17% for puerpera with height<145 cm, 4.83% for puerpera with age <20 years old, and 3.05% for puerpera with age ≥35 years old. The risks of low birth weight [OR (95%CI) value] of pregnant women aged <20, 25-29 and 30-34 years old were 1.31 (1.28-1.35), 0.88 (0.86-0.90) and 0.89 (0.87-0.91) times of those aged ≥35 years old. The risks of low birth weight [OR (95%CI) value] of pregnancy BMI <18.5 and 18.5-23.9 kg/m(2) group were 1.98 (1.94-2.03) and 1.20 (1.18-1.23) times of those pregnancy BMI ≥24 kg/m(2). The risks of low birth weight [OR (95%CI) value] of pregnant women's height (cm)<145, 145-154, 155-159 and 160-164 cm were 4.67 (4.39-4.97), 2.36 (2.29-2.44), 1.58 (1.53-1.63) and 1.22 (1.18-1.26) times of those heights ≥165 cm group. The risks of low birth weight [OR (95%CI) value] of pregnant women's gestational age <28, 28-31 and 32-36 years old were 136.65 (124.33-150.20), 1 704.37 (1 509.02-1 925.02) and 33.45 (32.98-33.94) times of those gestational age ≥37 years old. Conclusion: The incidence of low birth weight of live births was higher in Guangxi from 2016 to 2018. There is a higher risk of low birth weight for younger, older, low height, low BMI and preterm women in Guangxi from 2016 to 2018.

[Clinical observation on the anthracyclines-induced cardiotoxicity in patients with early-stage breast cancer].

Objective: To evaluate the anthracyclines-induced cardiotoxicity in patients with early-stage breast cancer. Methods: This retrospective study analyzed data of 64 patients (aged from 36 to 59 years old) with early-stage breast cancer after surgery. Patients were divided into ACT group (n=21), FAC group (n=19) and EC group (n=24). The NCI CTC 4.0 scores was used to evaluate the side effects at the time of 2 weeks, 4 weeks and 6 weeks after chemotherapy. Meanwhile, the level of cTnT, the incidence of abnormal electrocardiogram (ECG) and left ventricular ejection fraction (LVEF) were used to evaluate the anthracyclines-induced cardiotoxicity, the follow-up observation points were as follows: at the acute cardiotoxicity (time A), subacute cardiotoxicity (time B), 24 months after chemotherapy (time C), 36 months after chemotherapy (time D), 48 months after chemotherapy (time E), 60 months after chemotherapy (time F). The 3-years and 5-years overall survival and progress free disease survival among three groups were compared. Results: The ages, clinical stage, the size of tumor, axillary lymph node positivity and Eastern Cooperative Oncology Group Scores were similar among three groups (P>0.05); the incidence of side effects level 4 was 0. The levels of cTnT in the three groups were significantly lower than those at the baseline and time points C, D, E and F (all P<0.05), and the levels of cTnT were significantly higher in EC group than in FAC and ACT group at the time points B, C, D, E and F (P<0.05); however, the incidence of abnormal ECG and LVEF was similar among the 3 groups (P>0.05). The 5-year overall survival was 95.2% (20/21) ,100% (19/19) and 95.8% (23/24) in ACT group, FAC group and EC group, respectively; 5-year progress free disease survival was 95.2% (20/21) ,94.7% (18/19) and 91.7% (22/24) in ACT group, FAC group and EC group, respectively (P>0.05) . Conclusions: Patients with early-stage breast cancer after surgery could tolerate the anthracyclines-induced cardiotoxicity. Three chemotherapy schemes of ACT, FAC and EC, especially the EC protocol, could affect the myocardial damage. However, outcome is comparable among patients treated with above chemotherapy schemes in this patient cohort.

Comparison of 2015 Medicare relative value units for gender-specific procedures: Gynecologic and gynecologic-oncologic versus urologic CPT coding. Has time healed gender-worth?

BACKGROUND: In 1992, Congress implemented a relative value unit (RVU) payment system to set reimbursement for all procedures covered by Medicare. In 1997, data supported that a significant gender bias existed in reimbursement for gynecologic compared to urologic procedures. The present study was performed to compare work and total RVU's for gender specific procedures effective January 2015 and to evaluate if time has healed the gender-based RVU worth. METHODS: Using the 2015 CPT codes, we compared work and total RVU's for 50 pairs of gender specific procedures. We also evaluated 2015 procedure related provider compensation. The groups were matched so that the procedures were anatomically similar. We also compared 2015 to 1997 RVU and fee schedules. RESULTS: Evaluation of work RVU's for the paired procedures revealed that in 36 cases (72%), male vs female procedures had a higher wRVU and tRVU. For total fee/reimbursement, 42 (84%) male based procedures were compensated at a higher rate than the paired female procedures. On average, male specific surgeries were reimbursed at an amount that was 27.67% higher for male procedures than for female-specific surgeries. Female procedure based work RVU's have increased minimally from 1997 to 2015. CONCLUSION: Time and effort have trended towards resolution of some gender-related procedure worth discrepancies but there are still significant RVU and compensation differences that should be further reviewed and modified as surgical time and effort highly correlate.

Narcissus yellow stripe virus and Narcissus mosaic virus detection in Narcissus via multiplex TaqMan-based reverse transcription-PCR assay.

AIMS: Development of a multiplex TaqMan RT-qPCR assay to simultaneously detect Narcissus yellow stripe virus (NYSV) and Narcissus mosaic virus (NMV), frequently causing mixed narcissus infection. Feasibility verification was confirmed in natural samples. METHODS AND RESULTS: Primers and probes were designed based on the conserved CP gene regions of NYSV or NMV and their suitability for singleplex and multiplex TaqMan RT-qPCR assays as well as for conventional RT-PCR. Conventional RT-PCR, singleplex and multiplex TaqMan RT-qPCR assays proved to be NYSV and NMV specific. P-values and coefficients of variation of TaqMan RT-qPCR assays indicated high reproducibility. Significantly increased sensitivity was achieved compared to conventional RT-PCR. The detection limit of both viruses was 103 copies with superior correlation coefficients and linear standard curve responses between plasmid concentrations and Ct values. NYSV and NMV infection of narcissus leaves, petals and bulbs could successfully be detected via our multiplex RT-qPCR method at 1·25 mg. CONCLUSION: Our multiplex TaqMan RT-qPCR assay provides rapid, specific, sensitive and reliable testing to simultaneously detect NYSV and NMV, supplying useful routine monitoring for different narcissus samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Efficient identification and discrimination of the narcissus viruses provides reliable information for scientists and conventional growers. Furthermore, it enriches the information of NYSV, NMV and other narcissus viruses.

[Primary investigation of immunoglobulin A synthesis and secretion in human mesangial cells].

OBJECTIVE: To investigate the expression of immunoglobulin A (IgA) in human mesangial cells (HMCs). METHODS: The HMCs were cultured. The subcellular location of IgA was detected by immunofluorescence staining; the transcripts of Igα,Igκ and IgΛ constant region were detected by reverse transcription-polymerase chain reaction (RT-PCR) and further analyzed by DNA sequencing. The expressions of Igα and IgΛ were detected at transcription level by Western blot after the cytoplasmic protein extraction. The culture supernatant was collected to explore whether IgA could be secreted out of the cell and the protein was further analyzed by mass spectrometry after being purified by affinity chromato-graphy with jacalin-sepharose. The results of DNA sequencing and mass spectrometry were aligned with the mRNA and amino acid sequences in the National Center of Biotechnology Information (NCBI) database. RESULTS: By immunofluorescence staining, we detected the presence of IgA heavy chain Igα, light chain, both Igκ and IgΛ in expressions of transcripts of Igα1, Igα2, Igκ and IgΛ in the HMCs and the alignment of the sequences of the RT-PCR products with those of the IgCα1, IgCα2, Igκ and IgΛ mRNA in the NCBI database exhibited that the similarities were 99%, 97%, 98% and 97%, respectively. Western blot showed Igα and IgΛ expressions in the cell lysate and secretion of Igα1 and Igα2 heavy chains in cell culture supernatant. To further explore the protein that secreted into the supernatant, after supernatant affinity chromatography with jacalin-sepharose, the proteins were separated by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and the band approximating to 65 000 was cut and sent to mass spectrometry. The results were aligned with the amino acid sequences of Igα1 and Igα2 constant region in NCBI database, showing that amino acids between No.52 and No.104, amino acids between No.154 and No.221, amino acids between No.276 and No.327 from IgCα1 and amino acids between No.52 and No.113, amino acids between No.151 and No.204, amino acids between No.251 and No.314 from IgCα2 were the same with those derived from B cells. CONCLUSION: Our fin-dings suggested that HMCs could synthesize and secret IgA.

[Outcome determinants in cardiac arrest patients secondary to acute myocardial infarction receiving extra-corporeal membrane oxygenation combined with percutaneous coronary intervention therapy].

Objective: To investigate the clinical efficacy and outcome determinants in cardiac arrest patients secondary to acute myocardial infarction treated with extracorporeal membrane oxygenation (ECMO) and percutaneous coronary intervention (PCI). Methods: The clinical data of 27 patients hospitalized from January 2014 to March 2017 in 3 hospitals were retrospectively analyzed. The clinical data of the surviving group (12 cases) and the death group (15 cases) were compared and the outcome determinants were explored. Results: Twenty seven patients were successfully treated with coronary angiography and emergency PCI under ECMO assistance, and the successful procedure rate was 100%. The survival rate was 44.4% (12/27). There was no significant difference in gender, age, body weight, myocardial infarct location, past disease history and smoking status between the two groups (all P>0.05). Traditional cardiopulmonary resuscitation time was significantly longer, the CCU hospitalization time was significantly shorter, the number of diseased vessels was significantly higher, and the prevalence of distribution of blood vessels in left main stem was significantly higher and mean artery pressure at 24 and 48 hours post ECMO was significantly lower in the death group than in survival group (all P<0.05). Multiple logistic regression analysis showed that left anterior descending artery lesion, higher number of lesion vessels, longer traditional cardiopulmonary resuscitation time, longer time interval between cardiac arrest and ECMO placement were related increased risk of death post ECMO and emergency PCI in this patient cohort(OR=1.316, 95%CI 1.217-5.792, P=0.002; OR=1.238, 95%CI 1.107-4.961, P=0.000; OR=1.712, 95%CI 1.136-3.973, P=0.001; OR=1.629, 95%CI 1.132-4.521, P=0.000, respectively), while higher mean artery pressure at 48 hours post ECMO was related with reduced risk of death post ECMO and emergency PCI in this patient cohort(OR=0.672, 95%CI 0.326-0.693, P=0.001). Conclusions: ECMO combined with emergency PCI can improve the success rate of traditional cardiopulmonary resuscitation in patients with cardiac arrest secondary to acute myocardial infarction. Left anterior descending artery lesion, number of lesion vessels, traditional cardiopulmonary resuscitation time, time interval between cardiac arrest and ECMO placement and mean artery pressure at 48 hours post ECMO are outcome determinants post ECMO and emergency PCI in this patient cohort.

[Prevalence of human papillomavirus infection and risk of cervical cancer or precancerous lesions in 15 years follow up: a prospective cohort study].

Objective: To evaluate the 15 years changing trends of prevalence of high risk HPV (HR-HPV) infection and the risks of cervical cancer and precancerous lesions (CIN2+ ) among a Chinese rural population. Methods: The screening cohort with 1 997 women aged 35 to 45 years old was built in 1999 in Xiangyuan County, Shanxi province (SPOCCS-I) and followed up by cytology and HR-HPV testing in the years of 2005, 2010, and 2014. The changes of HR-HPV prevalence and the risks of cervical precancerous lesions with CIN2+ as the endpoints were analyzed during the past 15 years. Results: The detection rates of HPV infection and CIN2+ were 15.7%-22.3% and 1.1%-4.3% for the baseline visit and the other 3 follow-ups, respectively. The cumulative risk of CIN2+ in HR-HPV positive women at baseline was significantly higher than HR-HPV negative women (P<0.01) during the 15-year follow-up. The risk of CIN2+ in the four-times HPV positive group was 40.0%, while the group with four-times negative HPV results was 0.6% (Adjusted RR = 55.0, 95% CI: 11.3 to 268.4). Conclusions: The prevalence of HR-HPV infection and CIN2+ lesions were high in Xiangyuan county during the 15 years. HR-HPV positivity elevated the risk of CIN2+ compared to women whose HR-HPV test was negative. The risks of CIN2+ incidence in 6 years were low among women with negative HR-HPV test. The risk of CIN2+ increased with the numbers of HPV infection events. The screening interval could be extended to 5-6 years.

RNAi-mediated knockdown of relaxin decreases in vitro proliferation and invasiveness of osteosarcoma MG-63 cells by inhibition of MMP-9.

PURPOSE: The purpose of this study is to determine the role of relaxin knowdown by siRNA transfection in cellular growth and invasion of osteosarcoma MG-63 cells, and discusses the molecular mechanisms of this action. MATERIALS AND METHODS: The expression of relaxin in MG-63 cell was examined by western blot or RT-PCR. To evaluate the biological role of relaxin, proliferation assay (MTT) and invasion assay (BD Matrigel™), apoptosis assay (TUNEL and ELISA) and cell cycle analysis (flow cytometer) were performed after silencing relaxin using siRNA. MMP-9 expressions were analyzed using RT-PCR, western blot and zymography after silencing relaxin. RESULTS: Results showed that the downregulation of relaxin expression by siRNA in human osteosarcoma MG-63 cells significantly inhibited cell proliferation and invasion in vitro. Furthermore, relaxin knockdown led to cell arrest in the G1/G0 phase of the cell cycle, and eventual apoptosis enhancement in MG-63 cells. We provide evidence in our cell model that the relaxin siRNA down-regulated the expression of MMP-9 and the MMP-9 activity, suggesting that relaxin may promote the proliferation, invasion and metastasis of osteosarcoma cells by regulating the expression of MMP-9 and facilitating ECM degradation. CONCLUSIONS: Therefore, siRNA-directed knockdown of relaxin may represent a viable clinical therapy for osteosarcoma.

Relaxin promotes in vitro tumour growth, invasion and angiogenesis of human Saos-2 osteosarcoma cells by AKT/VEGF pathway.

OBJECTIVES: In the present study, we determine the role of relaxin on cellular growth, invasion and angiogenesis of osteosarcoma Saos-2 cells in vitro, and discuss the molecular mechanisms of this action. MATERIALS AND METHODS: Saos-2 cells were transfected with Akt1/2 siRNA or VEGF siRNA for 24 hours then treated with 10-100 ng/mL recombinant human relaxin-2 (rh-RLN) for 48 h. MTT, matrigel and bone marrow-derived endothelial cells (BMDECs) was used for cell proliferation, invasion and angiogenesis assay. Western blot was used for relaxin-2, pAKT and VEGF protein assay. RESULTS: The results showed treatment with 10-100 ng/mL rh-RLN resulted in 18%, 48%, 107%, 212% increase in cell proliferation, respectively (vs control, *p < 0.05;**p < 0.01), the relative invasive cells was 1.4;1.9;2.6;4.8 (control was defined to 1) (vs control, #p < 0.01; ##p < 0.001) and the relative anglogenic branch points in Saos-2 cells was 1.04;1.36;1.69;2.10 (control was defined to 1.00) (vs control, *p < 0.05; **p < 0.01). Furthermore, treatment with rh-RLN exhibited a significant increase in the expression level of pAKT and VEGF proterin in dose-dependent manner. Saos-2 cells were transfected with AKT1/2 siRNA for 24 h. No significant increase of VEGF protein expression was shown after rh-RLN treatment. CONCLUSIONS: These results suggested that rh-RLN could promoted proliferation, invasion and angiogenesis by upregulation pAKT-dependent VEGF expression.

Heterozygous Ins/Del genotype of the CASP8 rs3834129 polymorphism significantly decreases the risk of coal workers' pneumoconiosis in a Chinese Han population: a case-control study.

OBJECTIVE: Coal workers' pneumoconiosis (CWP) is a chronic inflammatory and fibrotic pulmonary disease that involves a complex interaction of multiple environmental and genetic factors. Polymorphism, as a genetic factor, may affect the onset of the disease in susceptible populations. The present study investigated the association between the polymorphisms of six genes and CWP risk in a Chinese Han population. PATIENTS AND METHODS: Six polymorphisms (CASP8 rs3834129, IL1A rs1800587, IL6 rs1800796, IL4 rs2070874, TNFA rs361525, and NLRP3 rs1539019) were examined in 222 CWP subjects and 247 dust-exposed control subjects. RESULTS: The CASP8 rs3834129 Ins/Del genotype significantly decreased CWP risk (p=0.040; adjusted odds ratio [OR] = 0.586; 95% confidence interval [CI] 0.367-0.935) compared with the Ins/Ins genotype. Stratification analyses revealed a significant interaction between the heterozygous Ins/Del genotype and age. Compared with the Ins/Ins + Del/Del genotype, this was particularly evident among subjects aged 41-60 (p<0.001; adjusted OR = 0.054; 95% CI 0.007-0.420) and those with an exposure time of 20-29 years (p=0.014; adjusted OR = 0.392; 95% CI 0.183-0.842). This decreased risk was also found in the group with former smokers (p=0.012; adjusted OR = 0.448; 95% CI 0.238-0.844). Findings revealed that the heterozygous Ins/Del genotype of CASP8 rs3834129 was related to a significantly decreased risk of stage I CWP (p=0.045; adjusted OR = 0.592; 95% CI 0.353-0.992), but not stage II or III CWP. CONCLUSIONS: Our study indicated that the heterozygous Ins/Del genotype of CASP8 rs3834129 significantly decreased CWP risk in a Chinese Han population.

Use of an expanded gold standard to estimate the accuracy of colposcopy and visual inspection with acetic acid.

We estimate the accuracy of colposcopy and visual inspection with acetic acid (VIA) while minimizing the effects of misclassification bias, and maximizing ascertainment of disease. VIA was performed by experienced physicians on a population-based sample of women aged 30 to 49 years in rural Shanxi province, China. Each woman received VIA, liquid-based cytology (LBC) and hybrid capture 2 (hc2, QIAGEN, Gaithersburg, MD; formerly Digene Corporation). Any woman who tested positive on any test had colposcopy, endocervical curettage (ECC) with directed biopsies as necessary and 4-quadrant random biopsies from normal-appearing areas of the cervix. A standard diagnosis based on colposcopy and directed biopsy, and an expanded diagnosis including ECC and 4-quadrant random biopsy were generated for each woman. In 1,839 women, use of the expanded versus the standard diagnostic criteria increased the prevalence of histologically confirmed high-grade cervical intraepithelial neoplasia and cancer (CIN2+) from 3.2% (59/1,839) to 4.2% (77/1,839) and decreased the sensitivity of VIA for CIN2+ from 69.5% (95% CI: 56.8-79.8) to 58.4% (95% CI: 47.3-68.8%) with little change in specificity of approximately 89%. Compared with the expanded diagnostic criterion, the sensitivity of a visual diagnosis of high-grade CIN or cancer by a colposcopist was 49.4% (95% CI: 38.2-60.5). The use of an expanded diagnostic criterion in this study yielded more conservative estimates of the sensitivity of VIA and colposcopy.

Immunohistochemical evidence for macroautophagy in neurones and endothelial cells in Alzheimer's disease.

AIM: To determine the pathological structures associated with macroautophagy in Alzheimer's disease (AD) and any relationship to disease progression. METHODS: Immunohistochemistry using antibodies to beclin-1, Atg5 and Atg12, early macroautophagy markers and LC3, the mammalian homologue of the later macroautophagy marker Atg8, were localized in formalin-fixed, paraffin-embedded medial temporal lobe sections of AD cases at variable neuritic disease stages. Double immunofluorescence labelling was used to co-localize these macroautophagy markers with Abeta and phospho-tau (AT8) and correlations performed using Spearman rank tests. RESULTS: Atg12 immunoreactivity in AD was either dispersed in the soma and dendrites or concentrated in tau-immunoreactive dystrophic neurites and some neurofibrillary tangles. Fewer Atg12-immunopositive neurones were observed with longer disease durations. Atg12-immunoreactive endothelial cells were found spatially associated with Abeta-positive plaques, with more Atg12-immunoreactive capillary endothelial cells with higher neuritic disease stage. These findings were confirmed by the other autophagy markers beclin-1, Atg5 and LC3. CONCLUSION: The data confirm that macroautophagy occurs in neurones undergoing neuritic degeneration in AD, identified early macroautophagy markers in capillary endothelial cells in close proximity to Abeta plaques, and found that evidence for macroautophagy changes with disease progression.

Arsenic uptake and metabolism in plants.

Arsenic (As) is an element that is nonessential for and toxic to plants. Arsenic contamination in the environment occurs in many regions, and, depending on environmental factors, its accumulation in food crops may pose a health risk to humans.Recent progress in understanding the mechanisms of As uptake and metabolism in plants is reviewed here. Arsenate is taken up by phosphate transporters. A number of the aquaporin nodulin26-like intrinsic proteins (NIPs) are able to transport arsenite,the predominant form of As in reducing environments. In rice (Oryza sativa), arsenite uptake shares the highly efficient silicon (Si) pathway of entry to root cells and efflux towards the xylem. In root cells arsenate is rapidly reduced to arsenite, which is effluxed to the external medium, complexed by thiol peptides or translocated to shoots. One type of arsenate reductase has been identified, but its in planta functions remain to be investigated. Some fern species in the Pteridaceae family are able to hyperaccumulate As in above-ground tissues. Hyperaccumulation appears to involve enhanced arsenate uptake, decreased arsenite-thiol complexation and arsenite efflux to the external medium, greatly enhanced xylem translocation of arsenite, and vacuolar sequestration of arsenite in fronds. Current knowledge gaps and future research directions are also identified.

A major quantitative trait locus controlling cadmium translocation in rice (Oryza sativa).

The trait of low cadmium (Cd) accumulation in brown rice (Oryza sativa) is important for food safety. An effective way to reduce Cd accumulation in the grain is to control Cd transfer from the roots to the shoots. Here, we investigated genotypic variation in the shoot Cd concentration among 146 accessions from a rice core collection and performed a quantitative trait locus (QTL) analysis to determine the loci controlling shoot Cd accumulation. Furthermore, we physiologically characterized the two accessions used for QTL analysis. Large genotypic variation (13-fold) in the shoot Cd concentration was found. A major QTL was detected on chromosome 11 using a F2 population derived from Badari Dhan (a high-Cd accession) and Shwe War (a low-Cd accession). This QTL explained 16.1% of the phenotypic variation in Cd accumulation. Furthermore, this QTL was confirmed by analysis of advanced progeny. Physiological studies showed that Badari Dhan and Shwe War did not differ in uptake of Cd by the roots, but differed greatly in the translocation of Cd from the roots to the shoots. Taken together, our findings suggest that the major QTL detected is responsible for the translocation of Cd from the roots to the shoots.