[Revision rate of periprosthetic joint infection post total hip or knee arthroplasty of 34 hospitals in China between 2015 and 2017: a multi-center survey].

Objective: To investigate the rate of periprosthetic joint infection (PJI) revision surgeries and clinical information of hip-/knee- PJI cases nationwide from 2015 to 2017 in China. Methods: An epidemiological investigation. A self-designed questionnaire and convenience sampling were used to survey 41 regional joint replacement centers nationwide from November 2018 to December 2019 in China. The PJI was diagnosed according to the Musculoskeletal Infection Association criteria. Data of PJI patients were obtained by searching the inpatient database of each hospital. Questionnaire entries were extracted from the clinical records by specialist. Then the differences in rate of PJI revision surgery between hip- and knee- PJI revision cases were calculated and compared. Results: Total of 36 hospitals (87.8%) nationwide reported data on 99 791 hip and knee arthroplasties performed from 2015 to 2017, with 946 revisions due to PJI (0.96%). The overall hip-PJI revision rate was 0.99% (481/48 574), and it was 0.97% (135/13 963), 0.97% (153/15 730) and 1.07% (193/17 881) in of 2015, 2016, 2017, respectively. The overall knee-PJI revision rate was 0.91% (465/51 271), and it was 0.90% (131/14 650), 0.88% (155/17 693) and 0.94% (179/18 982) in 2015, 2016, 2017, respectively. Heilongjiang (2.2%, 40/1 805), Fujian (2.2%, 45/2 017), Jiangsu (2.1%, 85/3 899), Gansu (2.1%, 29/1 377), Chongqing (1.8%, 64/3 523) reported relatively high revision rates. Conclusions: The overall PJI revision rate in 34 hospitals nationwide from 2015 to 2017 is 0.96%. The hip-PJI revision rate is slightly higher than that in the knee-PJI. There are differences in revision rates among hospitals in different regions.

[A ten-year follow-up study of a patient with chronic renal failure and metastatic pulmonary calcification after parathyroidectomy and review of the literature].

Objective: To analyze whether parathyroidectomy can prevent the progress of metastatic pulmonary calcification (MPC) in patients with chronic renal failure (CRF). Methods: A male patient with CRF complicated with MPC who underwent parathyroidectomy for secondary hyperparathyroidism and parathyroid adenoma was followed up for 10 years. The changes of MPC and the levels of blood calcium and phosphorus were measured. We searched the relevant literatures in PubMed and Wanfang databases with the key words of "metastatic pulmonary calibration" and "parathyroidectomy". Then, we manually retrieved the references of the literatures. A total of 18 patients (17 patients from 14 publications as well as the present case) were analyzed. By comparing the characteristics of MPC improvement group and MPC progression group, the factors affecting the prognosis of MPC after parathyroidectomy were explored. Results: After parathyroidectomy, the thoracic CT images of the patient gradually worsened from normal to diffuse ground glass opacity of both lungs, which indicated that parathyroidectomy did not prevent the progression of MPC in this patient. Among the 18 MPC patients who underwent parathyroidectomy, 10 patients had improved MPC, three had CRF, and two received peritoneal dialysis or hemodialysis respectively; eight patients had progressed MPC, all of the patients were CRF patients, one patient received peritoneal dialysis, and other patients received hemodialysis. Compared between the two groups, the proportion of CRF patients (P=0.004) and hemodialysis patients (P=0.003) in the progression group were significantly higher than those in the improvement group. Conclusion: Parathyroidectomy cannot prevent the progression of MPC in hemodialysis patients with CRF.

[Serial changes in biomarkers in pulmonary and extrapulmonary acute respiratory distress syndrome].

Objective: To compare the serial changes of circulating angiopoietin-2 (Ang-2), surfactant protein D (SP-D) and soluble receptor for advanced glycation end product (sRAGE) in patients with pulmonary acute respiratory distress syndrome (ARDS) and extrapulmonary ARDS. Methods: A total of 36 eligible patients with ARDS were included from the Department of Respiratory Intensive Care Unit(RCU) and Department of Intensive Care Unit (ICU), the Central Hospital of Wuhan, during April 2017 to February 2018, and divided into pulmonary ARDS group (n=20) and extrapulmonary ARDS group (n=16) according to primary causes. The levels of Ang-2, SP-D and sRAGE in serum of each group were measured on Day 1, 4 and 7 after diagnosis. Results: The serum level of Ang-2 in extrapulmonary ARDS group declined gradually during the course. The serum level of Ang-2 on Day 1 was significantly higher than that on Day 7 [2 801 (1 386, 7 526) vs 1 461 (737, 2 135) ng/L, P<0.05]. There was no significant difference in serum levels of Ang-2 on Day 1, 4, 7 in pulmonary ARDS group [1 462 (545, 3 715) vs 1 353 (659, 5 847) vs1 616 (754, 3 460) ng/L, all P>0.05]. There was no significant difference in serum levels of SP-D on Day 1,4, 7 in each group [pulmonary ARDS group: 5.6(1.8, 14.0) vs 5.7(1.8, 14.5) vs 7.9(3.0, 16.8) mg/L, all P>0.05; extrapulmonary ARDS group: 3.4(0.5, 9.1) vs 1.6(0.6, 6.3) vs 1.6(0.7, 7.9)mg/L, all P>0.05]. The serum level of sRAGE in each group declined gradually during the course. The serum level of sRAGE on Day 1 in each group was significantly higher than that on Day 7 [pulmonary ARDS group: 328(107, 595) vs 66 (50,171) ng/L, extrapulmonary ARDS group: 237 (98, 410) vs 81 (38, 154) ng/L, all P<0.05]. Conclusion: The serum level of Ang-2 in extrapulmonary ARDS declines significantly in the early stage of disease, while there is no significant difference in that of pulmonary ARDS.

[Effect of obstructive sleep apnea on the severity of acute pulmonary thromboembolism].

Objective: To investigate the effect of obstructive sleep apnea (OSA) on the severity of acute pulmonary thromboembolism (PTE). Methods: Clinical data of patients with acute PTE and OSA who were admitted to Anzhen Hospital from January 2015 to December 2017 were retrospectively analyzed in this study.According to the apnea hypopnea index (AHI), patients were divided into AHI≤15/h group and AHI>15/h group. The levels of oxygen desaturation index (ODI), mean pulse oxygen saturation (MSO(2)), the lowest pulse oxygen saturation (LSaO(2)), oxygen saturation<90% time ratio (Ts90%), hemoglobin, hematocrit (HCT), blood platelet, mean platelet volume (MPV), platelet distribution width (PDW), hypersensitive C-reactive protein (hsCRP), homocysteine (HCY), prothrombin time, activated partial thromboplastin time (APTT), fibrinogen (FIB), D-dimer, fibrin degredation product (FDP), B-type natriuretic peptide, cardiac troponin I (cTnI), arterial partial pressure of oxygen (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)) were compared between AHI≤15/h group and AHI>15/h group. Differences in the severity of PTE between the two groups were compared; binary Logistic regression model was used to analyze the risk factors of acute PTE severity. Results: A total of 75 patients with acute PTE and OSA were enrolled. Patients in the AHI>15/h group were significantly older [(68.7±9.9) vs (62.8±12.8) years old, P=0.029], and were more likely to have chronic heart failure and/or lung disease (53.1% vs 23.1%, P=0.012) and elevated Pulmonary Embolism Severity Index scores (61.2% vs 30.8%, P=0.012) than those in the AHI≤15/h group. Compared to the AHI>15/h group, the levels of D-dimer and B-type natriuretic peptide were much lower in the AHI ≤15/h group [243.0 (140.0, 471.5) vs 408.0 (258.0, 1 009.5) µg/L, P=0.01; 48.0 (19.8, 87.5) vs 70.0 (34.5, 201.0) ng/L, P=0.039], while arterial oxygen partial pressure was significantly higher [(79.0±10.6) vs (73.4±8.2) mmHg (1 mmHg=0.133 kPa), P=0.015]. In patients with acute PTE and OSA, AHI (P=0.030) and B-type natriuretic peptide level (P=0.023) were independently associated with an increased risk of acute PTE severity. Conclusion: In patients with PTE and OSA, moderate or severe OSA may aggravate the severity of acute PTE.

[Concordance of PD-L1 expression detected by two immunohistochemical platforms and antibodies].

Objective: To evaluate the concordance of PD-L1 expression in various tissues using antibodies 28-8 and SP263 on their respective detection platforms. Methods: Three hundred seventy four specimens of surgical resection of pulmonary diseases in the First Affiliated Hospital of Nanjing Medical University from January 1, 2012 to January 31, 2017 were collected. Totally 374 cases were tested for PD-L1 expression using the two antibodies, 28-8 and SP263, by respective detection platforms (Dako and Ventana). Finally, 336 cases were used for further evaluation, and the results were statistically analyzed for concordance. Results: For non-small cell lung carcinoma (NSCLC), the positive rate of PD-L1 was 57.5% (177/308) using SP263, and 57.5% (177/308) using 28-8 antibody. The correlation coefficient was 0.97 (P<0.01). The positive rate of both benign lung diseases and paracancerous tissues was about 10.7% (3/28), and the positive concordance rate was 100.0%. The distribution of both antibodies was also relatively consistent. Conclusions: The expression levels of 28-8 and SP263 antibodies in NSCLC and other tissues are relatively consistent, suggesting both antibodies may be complementary and substitute for each other, which may be useful in guiding clinical management.

Baicalin inhibits Escherichia coli isolates in bovine mastitic milk and reduces antimicrobial resistance.

In this study, we aimed to evaluate the inhibitory effect of baicalin on Escherichia coli in vitro and the effects of baicalin treatment on antimicrobial resistance of the E. coli isolates. Through isolation, purification, and identification, a total of 56 E. coli strains were isolated from 341 mastitic milk samples. The study of inhibition effect of baicalin on the E. coli strains in vitro was focused on permeability and morphology of the isolates using an alkaline phosphatase kit and scanning electron microscopy. Furthermore, the resistance spectrum of the isolates to the common antimicrobial agents was tested at sub-minimum inhibitory concentrations of baicalin by the agar dilution method. Extended-spectrum ß-lactamase and plasmid-mediated quinolone resistance genes were amplified by PCR before and after incubation with baicalin. The results revealed that baicalin has certain inhibitory effects on the isolates in vitro. The alkaline phosphatase enzyme activity was significantly increased from 1.246 to 2.377 U/100 mL, and the surface of E. coli was concave and shriveled. Analysis of the resistance spectrum and PCR amplification showed that, after administration with baicalin, the sensitivity of most strains to the selected antimicrobial agents was enhanced. Strikingly, the drug-resistant genes from 71.43% (40/56) of these isolates were found to have drug-resistant genes to different extents. Altogether, the current study confirmed both the inhibitory effect on Escherichia coli in vitro and the reduction of antimicrobial resistance by baicalin. This is the first comprehensive study to report on baicalin, a traditional Chinese medicine that acts on E. coli isolated from the mastitic milk samples.

[The study on behaviour of protective equipment utilization of workers exposed to benzene and factors based on Planned Behavior Theory].

Objective: To investigate and predict the behavioral intention and mode of the protective equipment utilization selection of the workers who used Benzene, the Theory of Planned Behavior (TPB) was applied to establish the behavioral model to enhance the theoretical foundation for long-term intervention. Methods: Questionnaires were used to survey the 707 workers, and all the behaviors of using protective equipment were investigated. Evaluate the relationships between each variable and obtain the influence affects by structural equation model. Results: The investigation showed that 38.47% of the total workers (272 cases) used whole body protection, 13.58% used partially, and 16.69% didn't use any body protection. There were significant difference between the varying degrees in the four dimensions (behavioral attitude, perceived behavior control, subjective norm, and behavioral intention) (P<0.01) . The results of structural equation model revealed that perceived behavior control was the most important influencing factor, subjective norm, positive attitude, negative attitude were the other three respects in sequence. The path co-efficient were 0.600、0.215、0.141 and 0.046 respectively. Conclusion: The study show that the theory of planned behavior can effectively explain the behavioral intention and behavior of protective equipment utilization. Therefore, combining the subjective initiative of individuals with the supervision of enterprises, In order to effectively enhance the protective equipment utilization of benzene workers.

[Clinicopathologic features of mammary microglandular adenosis with carcinoma: a study of 5 cases].

Objective: To study the clinicopathologic, immunohistochemical features, differential diagnoses and prognosis of mammary microglandular adenosis with carcinoma (MGACA) with micropapillary pattern. Methods: Five cases of MGACA were collected from 2010 to 2016 and reviewed for their clinical, histologic features and outcome.EnVision method were done for S-100 protein, cytokeratin (CK), p63, Calponin, smooth muscle myosin heavy chain (SMMHC), PR, ER and HER2. Results: Histologically, microglandular adenosis(MGA), atypical MGA (AMGA) and invasive carcinoma were seen in all five cases of MGACA. The invasive component was metaplastic carcinoma in one case and ductal in four cases. All epithelial cells were S-100 and CK positive in MGA, AMGA and invasive carcinoma. p63, Calponin and SMMHC negativity confirmed the lack of a myoepithelial cell layer in MGA, AMGA and MGACA. PR was weakly focally positive in one case, but ER and HER2 were negative in all cases (four cases were triple negative). Ki-67 index was 20% to 40%. Laminin and collagen Ⅳ staining showed the presence of basement membrane in MGA and AMGA, except MGACA. The follow-up time ranged from 3 months to 6 years, and all patients were alive without recurrence or distant metastasis. Conclusions: MGACA is a rare tumor with distinct morphological and IHC features. Compared to most triple-negative breast cancers, MGACA seems to have a relatively favorable outcome.

[Correlation analysis of PD-L1 expression and prognosis in triple-negative breast cancers].

Objective: To investigate the relationship between PD-L1 expression and the clinicopathologic features and prognosis in triple-negative breast carcinomas (TNBC). Methods: All 142 cases of TNBC were collected from the First Affiliated Hospital of Nanjing Medical University from February 2011 to December 2014, and the surgical excision or biopsy specimens from patients without chemotherapy and radiotherapy were included. Histopathologic analysis of stromal tumor infiltrating lymphocyte (sTIL) was performed on HE sections, and PD-L1 immunohistochemical staining was done with MaxVision. Results: The PD-L1 expression rate was 34.5% (49/142) in tumor cells, and was 62.0% (88/142) in sTIL. The PD-L1 expression in tumor cells was positively correlated with tumor size (r=0.181, P=0.031), Ki-67 index (r=0.211, P=0.012), sTIL (r=0.380, P<0.01) and PD-L1 expression in sTIL (r=0.447, P<0.01). The PD-L1 expression in sTIL was positively correlated with tumor grade (r=0.215, P=0.01), Ki-67 index (r=0.253, P=0.002) and sTIL (r=0.370, P<0.01). The high stromal CD8(+) /FOXP3(+) ratio was significantly associated with improved overall survival (χ(2)=4.186, P=0.041). The high percentage of sTIL was significantly associated with improved overall survival (χ(2)=12.427, P<0.01) and progression-free survival (χ(2)=4.057, P=0.044). Conclusions: In TNBC, PD-L1 expression is positively correlated with Ki-67 and sTIL; the stromal CD8(+) /FOXP3(+) ratio and sTIL are significantly associated with prognosis. The PD-L1 expression, stromal CD8(+) /FOXP3(+) ratio and sTIL are biologically important in TNBC, and all these correlative factors are important potential parameters in assessing immunotherapy for TNBC.

[Prognostic study of visceral pleural invasion by pulmonary adenocarcinoma with tumor size ≤3 cm].

Objective: To evaluate the relationship between visceral pleural invasion (VPI) and other clinicopathological features in lung adenocarcinoma with tumor size ≤3 cm, and to investigate the impact of VPI on the patients' prognosis. Methods: The clinical and pathological features were retrospectively reviewed in 231 cases of lung adenocarcinoma with the largest diameter of tumor ≤3 cm, following complete resection and systemic lymphadenectomy. VPI was divided into three grades, PL0, PL1 and PL2 according to modified Hammar classification for lung cancer upon elastic fiber staining. Survival analysis was performed by Kaplan-Meier method, and the risk factors for prognosis were explored by Cox proportional hazards model. Patient prognosis was evaluated by progression-free survival (PFS) and overall survival (OS). Results: In all 231 cases, the number of patients with VPI was 70 (30.3%), of which 61 cases were PL1 and 9 cases were PL2. The remaining 161 cases (69.7%) had no VPI (PL0). The tumor size (P=0.003), histological grade (P<0.01), the presence of solid component (P=0.001) and micropapillary component (P=0.009), N stage (P<0.01) and TNM stage (P<0.01) were significantly correlated with VPI. Patients with VPI had significantly shorter PFS and OS than those without VPI (P<0.01). There were significant differences in PFS and OS between patients with different VPI levels (P<0.01). Cox multivariate regression analysis showed that VPI was not an independent prognostic factor, whereas PL2 was an independent prognostic factor for PFS (P=0.007), but not an independent prognostic factor for OS (P=0.052). Conclusions: For patients with lung adenocarcinoma of tumor size ≤3 cm, VPI is related to poor prognosis; However, only PL2 is an independent prognostic factor for PFS. It may be not necessary to separate PL0 and PL1 status in smaller lung adenocarcinomas. Therefore, the definition of VPI may need further modification through large cohort studies.

Metastasis-associated gene 1 expression in human medulloblastoma and its association with invasion and metastasis in medulloblastoma Daoy cell lines.

This study aims to investigate the expression of metastasis-associated gene 1 (MTA1) in human medulloblastoma, and its significance in the invasion and metastasis in a medulloblastoma cell line. Positive expression rate of MTA1 protein in medulloblastoma and adjacent normal tissues collected from 29 medulloblastoma patients was detected by immunohistochemistry assay in vivo. In in vitro experiments, Daoy cells were transfected with MTA1-targeted small interfering RNA (siRNA, MTA1-siRNA group), niRNA (MTA1-niRNA group), and plasmid vectors (control group). Transfection efficiency was evaluated by PT-PCR and western blot; cell adhesion, migration, and invasion capacity was assessed by adhesion assays, scratch assays, and transwell chamber invasion assays, respectively. Results indicated that the positive expression rate of MTA1 protein in the medulloblastoma tissues was higher as compared with that of the adjacent normal tissues (P < 0.05). In addition, mRNA and protein expression of MTA1 in the MTA1-siRNA group was lower than that in the control and MTA1- niRNA groups (P < 0.05). Adhesion, migration, and invasion capacity of Daoy cells in the MTA1-siRNA group was inhibited as compared with the control and MTA1-niRNA groups (P < 0.05). In conclusion, MTA1 expression was increased in medulloblastoma cells, while MTA1 knockdown in medulloblastoma cells inhibited MTA1 expression. In addition, MTA1 knockdown inhibited the adhesion, migration, and invasive capabilities of medulloblastoma cells. It is possible that MTA1 can serve as a biomarker and a potential therapeutic target for medulloblastoma.

[CD8 and FOXP3 expression in stromal tumor-infiltrating lymphocytes of triple-negative breast carcinomas: a clinicopathologic study].

OBJECTIVE: To investigate the relationship between stromal tumor-infiltrating lymphocytes (TIL) and CD8 or FOXP3 positive lymphocytes in triple-negative breast carcinoma. METHODS: A total of 160 triple-negative breast carcinomas were collected from the First Affiliated Hospital of Nanjing Medical University during 2012 to 2014. All were surgical excision or biopsy specimens from patients without prior chemotherapy and radiotherapy. Histopathologic analysis of stromal TIL was performed on hematoxylin and eosin-stained sections, and MaxVision immunohistochemical method was used for detection of CD8 and FOXP3 protein expression. RESULTS: Stromal TILs were positively correlated with Ki-67 labeling index (P=0.002). The density of CD8(+) cytotoxic T-lymphocyte (CTL) was negatively correlated with tumor size (P=0.009), and positively correlated with Ki-67 index (P=0.021). The density of FOXP3(+) regulatory T-lymphocyte (Treg) was inversely correlated with the patient age (P=0.030), and positively correlated with histological grade (P=0.026). Stromal TILs were positively correlated with the density of CD8(+) CTL (P=0.014). CONCLUSIONS: The percentage of stromal TILs and density of CD8(+) CTL are associated with tumor cell proliferation of triple-negative breast cancers. The density of FOXP3(+) Treg is significantly associated with poor prognosis. Stromal TIL is positively correlated with the density of CD8(+) CTL. Stromal TIL may provide a potential marker for pathological diagnosis and a target for guiding adjuvant therapy in patients with triple-negative breast cancers.

First Report of Greenhouse Tomato Wilt Caused by Plectosphaerella cucumerina in China.

Tomato (Lycopersicon esculentum Mill.) is one of the most important vegetable crops in China. In May 2011, root rot and plant wilt were observed on tomato plants (variety Jinguan No. 5 and Meina) in 26 commercial greenhouses in Huludao city, Liaoning Province, China. Disease incidence was 30 to 95%. At beginning of fruit set, symptoms were chlorosis of lower leaves and lack of turgidity in young leaves. Severely affected plants were wilted and stunted as fruit approached maturity. Primary and secondary roots became necrotic with few fine feeder roots. Symptomatic roots were collected and cut into small pieces, disinfested in 2% sodium hypochlorite for 2 min, rinsed with sterile water, and placed on potato dextrose agar. After incubation at 25°C for 5 days, 20 axenic cultures were obtained from single conidia. Colonies were buff or salmon pink, moist, and had appressed, slimy mycelium. Aerial mycelium was sparse with simple or branched conidiophores. Conidia were 4.0 to 8.9 × 2.0 to 4.0 (average 6.9 × 2.8) µm, aggregated in slimy heads, hyaline ellipsoidal and ovoid, smooth, and 0 to 1 septate. Conidia were borne on phialides that were 6.3 to 24.3 × 1.4 to 3.3 (average 15.9 × 2.2) µm. These characteristics are typical of Plectosphaerella cucumerina (Lindf.) W. Gams (1) (anamorph: Fusarium tabacinum) (3). The internal transcribed spacer (ITS) region of the 20 isolates was amplified using the primers ITS1/ITS4 and sequenced. Identical sequences were obtained from all 20 isolates, and the sequence of isolate HLDT15 was submitted to GenBank (Accession No. KC894931). BLAST analysis of the sequence showed 100% similarity to P. cucumerina (AB469880). Pathogenicity tests were conducted with tomato variety Jinguan No. 5. Six 12-liter pots were filled with sterilized potting mix (equal parts sand, peat, and soil) and 200 ml conidial suspension (1 × 105 conidia ml-1). The conidial suspension of the isolate of P. cucumerina was prepared from 7-day-old cultures grown in potato dextrose broth on a shaker (120 rpm) at 25 ± 1°C. Six control pots were filled with potting mix and 200 ml of sterilized potato dextrose broth. Each pot was sown with six surface-sterilized (2% sodium hypochlorite for 2 min) tomato seeds. All the pots were kept in a greenhouse at 23 to 28°C. Three to four weeks after seedling emergence, all inoculated plants were dwarfed and lower leaves were chlorotic. Roots were necrotic and produced fewer fibrous roots. All characteristics were similar to original observations on the host of origin. Control plants remained asymptomatic. The same results were obtained when pathogenicity tests were repeated twice. P. cucumerina was reisolated from inoculated plants and matched the morphological and molecular characteristics of the original isolates. P. cucumerina was reported as a pathogen of tomato in Italy (2) and Australia (4). To our knowledge, this is the first report of P. cucumerina causing tomato wilt in China. So far, we have observed the disease on tomatoes in commercial greenhouses of Pulandian and Panjin city, Liaoning Province, China. The spread of this disease may pose a threat to tomato production in China. References: (1) A. Carlucci et al. Persoonia 28:34, 2012. (2) A. Matta et al. Riv. Patol. Veg. 14:119, 1978. (3) M. E. Palm et al. Mycologia, 87:397, 1995. (4) I. G. Pascoe et al. Mycol. Soc. 83:343, 1984.

Validation of quantitative fluorescent-PCR for rapid prenatal diagnosis of common aneuploidies in the Chinese population.

Quantitative fluorescent polymerase chain reaction (QF-PCR) is an accurate and reliable method for rapid detection of aneuploidy; however, it is not routinely used in China. We aimed to validate QF-PCR as a means for prenatal common aneuploidy screening and to analyze the heterozygosities of short tandem repeat (STR) markers in the Chinese population. The sequences of 19 STR markers in chromosomes 21, 18, 13, X, and Y were designed; three kinds of fluoresceins were used to label the primers, and the QF-PCR detecting conditions were explored and optimized. The results of analysis of 210 prenatal samples by multiplex QF-PCR were compared with karyotyping analysis. All cases were successfully tested by QF-PCR and conventional cytogenetic analysis. QF-PCR results were consistent with the results of cytogenetic analyses, with the exception of two cases. The sensitivity and specificity of QF-PCR to diagnose common aneuploidies were 94.74 and 100%, respectively. The heterozygosities of most of the markers were lower than reported for Western populations, but relatively similar to those of other Asian populations. We conclude that QF-PCR is able to detect the common aneuploidies for prenatal diagnosis with high detection efficacy; therefore it is suitable for rapid prenatal diagnosis and for large-scale testing in laboratories. However, we need to add new STR markers or to find alternative STR markers with high heterozygosity in order to make this technique useful for routine diagnosis.

Effect of miR-133 on apoptosis of trophoblasts in human placenta tissues via Rho/ROCK signaling pathway.

OBJECTIVE: The aim of this study was to explore the role of micro ribonucleic acid (miR)-133 in the apoptosis of human placental trophoblasts through the Ras homolog gene family (Rho)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway. PATIENTS AND METHODS: The plasma samples were collected from 30 patients with pre-eclampsia (PE) undergoing treatment and 30 healthy subjects (control group) who received physical examination in our hospital. The Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was utilized to measure the expression of miR-133 in PE patients and healthy people. Meanwhile, blood pressure, urine protein content, liver function, and kidney function were detected in patients of both groups as well. Subsequently, the placental trophoblasts were extracted and transfected with inhibitors and miRNA mimics to suppress and overexpress miR-133, respectively. The transfection efficiency was determined by RT-PCR. The levels of interleukin-6 (IL-6), IL-1, and tumor necrosis factor-alpha (TNF-α) were measured in both groups. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay was performed to determine the apoptosis of trophoblasts. Next, the RT-PCR and Western blotting were carried out to detect the expressions of the Rho/ROCK pathway. Furthermore, the influence of miR-133 on the apoptosis of trophoblasts in human placenta tissues through Rho/ROCK was comprehensively observed. RESULTS: In vivo experiments demonstrated that the urinary protein content, miR-133 level, systolic blood pressure, diastolic blood pressure, and liver function and renal function indexes were significantly elevated in pre-eclampsia (PE) patients in comparison with normal subjects (p<0.05). After transfection of mimics and inhibitors, the expression of miR-133 was remarkably up- and down-regulated, respectively. The content of the inflammatory factors in miR-133 mimics group was overtly higher than the other two groups. The TUNEL staining results showed that the number of apoptotic cells significantly increased and decreased in the miR-133 mimics group and miR-133 inhibitors group, respectively. Subsequent experiments indicated that the expressions of apoptosis gene Caspase3, pathway gene, and protein ROCKI were notably up-regulated in miR-133 mimics group. However, they were evidently down-regulated in miR-133 inhibitors group than in the control group. In addition, a consistent trend was observed in the protein expression level. CONCLUSIONS: MiR-133 participates in the development and progression of PE through the Rho/ROCK signaling pathway, which may affect the apoptosis of trophoblasts in the placenta tissues.