KRAS mutations analysis in mucinous epithelial lesions of the endometrium / 中华病理学杂志

Objective@#To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features.@*Methods@#The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups.@*Results@#The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05).@*Conclusion@#The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.

Clinical application of STR genotyping diagnosis for hydatidiform mole and nonmolar gestation / 中华病理学杂志

Objective@#To investigate the value of short tandem repeat (STR) genotyping in the diagnostic workup of molar and non-molar gestations with correlation of histological characteristics.@*Methods@#Six hundred and fifty-six cases were selected based on clinically suspected hydropic abortion and/or molar pregnancy from July 2015 to September 2017 at Beijing Obstetrics and Gynecology Hospital. DNA was extracted from dissected chorionic villi and paired maternal endometrial FFPE tissue samples by Simplex OUP™ FFPE DNA Tissue Kit. STR genotyping was performed by PowerPlex 16 HS system.@*Results@#DNA genotyping was informative in 649 of 656 cases, leading to identification of 215 hydatidiform mole gestations and 434 non-molar gestations. Most of non-molar gestations (375 cases, 86.4%) were diploid hydropic abortion. Various trisomy syndromes were found (53 cases, 12.2%), including trisomy 2, 3, 4, 7, 8, 13, 16 and 21. Only 2(0.5%) digynic triploid gestations were detected. Moreover, 4 cases (0.9%) of uniparental disomies (homologous or heterologous) were found. There were 196 cases with histologic diagnostic suspicious of hydatidiform moles were accurate sub-classified. Among them, 59 cases hydatidiform moles were under-diagnosed as diploid hydropic abortions, and 28 cases diploid hydropic abortions were over-diagnosed as hydatidiform moles.Compared with partial moles(PHM), there were no specific histomorphological features between the various types of non-molar gestations and partial moles for definitive diagnostic separation. There was no significant difference in the expression of p57kip2 among PHM, trisomy and diploid hydropic abortions group (P=0.247).@*Conclusions@#STR genotyping can distinguish non-molar gestations from early hydatidiform moles, and efficiently avoid misdiagnosis based only on histological evaluation. Therefore, using STR genotyping, not only can the overdiagnosis of non-molar pregnancy be avoided, but also individualized management can be offered to patients including monitoring of serum hCG.

A multi-center study on realtime polymerase chain reaction assay for group B Streptococcus in pregnant ;women / 中华围产医学杂志

Objective To evaluate the accuracy of realtime polymerase chain reaction (PCR) assay in the detection of group B Streptococcus (GBS) in pregnant women. Methods Samples were collected from 1 395 women at 35-37 weeks of gestation from March 1 to December 31, 2009 at three hospitals in Beijing. Samples were obtained from the lower one third vaginal wall and perianal area and tested for GBS using standard culture and PCR. Standard culture and gene analysis for GBS were applied as the gold standard, and the sensitivity and specificity of the rapid assay were determined. Results Of the 1 395 women qualified for PCR testing, 40(2.9%) were identified as GBS positive on the basis of the results of specimen culture, compared to 114 (8.2%) on the basis of PCR assay. The culture was negative and the PCR positive in 77 patients. The results which were not in agreement using the two tests were evaluated by the gene analysis for GBS. Among the 77 samples which were GBS positive by PCR, 66 samples were determined as GBS positive by gene analysis. The sensitivity of the PCR assay was 97.2%(103/106) and specificity was 99.1%(1 278/1 289), the maternal GBS colonization was 7.6%(106/1 395). Conclusions Realtime PCR assay allows rapid and reliable detection of GBS in last trimester with high sensitivity and specificity.

Investigation on clinical efficacy of the fourth-generation ultrasonic lithotripsy system in the treatment of urinary tract stones / 中国基层医药

ObjectiveTo evaluate the fourth-generation ultrasonic lithotripsy system for treating the urinary tract stones.Methods243 cases of urinary tract stones who received treatment of fourth-generation EMS ultrasonic lithotripsy system were analyzed retrospectively.ResultsImmediate phase I lithotrotrisy was performed successfully in 227 cases and the successful rate of phase I was 93.3％ (227/243).Delayed phase Ⅱ lithotripsy was performed for 16 patients.Complications happened in 2 cases,one case occurred hydropneumothorax during operation,required indwelling thoracic drainage tube processing,the other had severe bleeding which conservative treatment was ineffective,cured by the intervention of super-selective renal artery embolization.Conclusion Fourth generation of ultrasonic lithotripsy for treating stones on different anatomical locations of urinary system was safe,practical and efficient,worthy of clinical application.

Real-time fluorescence PCR-molecular beacon assay for detecting mutations in rpoB genes of rifampin-resistant Mycobacterium tuberculosis / 中华检验医学杂志

Objective To establish a rapid method to detect mutations in rpoB genes of rifampin-resistant Mycobacterium tubereulosis in dinical specimens using Real-time fluorescence PCR molecular beacon assay.Methods 174 strains of Mvcobacterium tuberculosis clinical isolates were analyzed using real-time fluorescence PCR molecular beacon assay foilowed with DNA sequencing while 12 strains of NTM and 4 strains of bacteria other than Mycobacterium tuberculosis were used as the contrast.Results Eighty-two 89.1 of 92 rifampin (RIF)-resistant strains and 3 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using real-time fluorescence PCR-molecular beacon assay.The specificity, sensitivity,and accuracy of this assay were 96.3%,89.1%,and 92.5%,respectively-Eithty-three of 92 RIF-resistant strains and 1 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using the direct DNA sequencing.The specificity,sensitivity,and accuracy of the direct DNA sequencing were 98.8,90.2%,and 94.2%,respectively.As compared with real-time PCR molecular beacon assay,171 of 174(98.3%)strains of myeobactefium tuberculosis clinical isolates had the salne results.Conclusion Real-time fluorescence PCR-molecular beacon assay can be used as a rapid screen method to detect RIF-resistant isolates.

Establishment and characteristics of orthotopically transplanted model of human primary malignant spleen lymphoma in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To establish three orthotopically transplanted model of human primary malignant spleen lymphoma in the nude mice.</p><p><b>METHODS</b>Orthotopic transplantation of histologically intact human primary malignant splenic lymphoma tissue obtained from patients was introduced into the splenic parenchyma of nude mice. Tumorigenicity, invasion, metastasis and morphological characteristics of the transplanted tumor were studied by light microscopy, electron microscopy and immunohistochemical methods.</p><p><b>RESULTS</b>The first kind, a strain of human primary malignant spleen lymphoma (non-Hodgkin's, cleaved B cell, BFNHL-HMN-1) screened from 11 patients which had been passaged in vivo for 41 generations, a second kind, a liver metastasis model of human primary malignant spleen lymphoma (non-Hodgkin's, cleaved B cell, LM-BFNHL-HMN-2) which had been passaged for 47 generations and a third kind of human primary malignant spleen lymphoma (non-Hodgkin's, T-immunoblastic cell, TINHL-HMN-3) having passaged for 37 generations were all successfully transplanted in 611 nude mice. Models of BFNHL-HMN-1 and TINHL-HMN-3 tumor gave nodular growth and lymph node metastasis in the spleen hilum but without any metastasis in the abdominal lymph nodes or organs. In the LM-BFNHL-HMN-2 model, not only did the tumor cells grow in the spleen, but in spleen hilum, lymph nodes and liver also. The orthotopically transplanted tumor cells were similar to the original human tumor in light histopathology, ultrastructure features, DNA content and chromosomal karyotype.</p><p><b>CONCLUSION</b>These three models are able to serve as useful tools for the study of biologic characteristics and experimental treatment of human primary malignant lymphoma.</p>

Significance of HER-2/neu expression in ovarian epithelial tumours / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate expression of Her-2/neu (c-erbB-2) and its significance in ovarian epithelial tumors.</p><p><b>METHODS</b>106 specimens and clinical history of ovarian epithelial tumors were collected including 54 cases of malignant, 33 cases of borderline and 19 benign cases. There were 19 cases of stages I and II and 35 cases of stages III and IV in ovarian carcinoma and all borderline malignant cases were stage I according to FIGO's standard. Immunohistochemical staining for Her-2/neu was performed by LSAB method.</p><p><b>RESULTS</b>The positives rates of Her-2/neu of benign, borderline and malignant tumors were 47.4% (9/19), 84.8% (28/33,), and 85.2% (46/54), respectively. Both borderline and malignant cases were found to have higher expression of Her-2/neu than those of benign cases (P < 0.02 and P < 0.01) respectively. Overexpression of Her-2/neu showed in 14/54 (25.9%) of malignant cases, 3/33 (9.1%) of borderline, and none of benign cases. Overexpression of Her-2/neu in malignant tumors cases with metastasis was higher than those without metastasis (P < 0.001).</p><p><b>CONCLUSION</b>Overexpression of Her-2/neu is associated with biological aggressiveness of ovarian cancer.</p>

Monocyte chemoattractant protein-1 mRNA in human intracranial aneurysm walls / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the pathological course in intracranial aneurysms.</p><p><b>METHODS</b>Normal intracranial artery tissue (cortex fistulization) from 1 case, ruptured aneurysms tissuses from 11 cases, unruptured aneurysm tissues from 2 cases were obtained by neurosurgical excision. Routine HE staining was used to observe histological characteristics. In situ hybridization was used to observe the expression of the monocyte chemoattractant protein-1 (MCP-1) mRNA in the walls of the normal artery and aneurysms.</p><p><b>RESULTS</b>By the HE staining showed that the wall of the ruptured aneurysms (10 cases) and unruptured ones (2 cases) had increased intima and connectivum extima. The fibroblast in the intima was arrayed in the disorder. Monocyte-like cells can be seen in the whole aneurysm wall. In one case aneurysms wall (ruptured) glass-like fiber structure was left over, few cells could be seen. In 9 cases, mural thrombus was found. The thrombus represented with organization. In situ hybridization, MCP-1 mRNA was not detectable in the normal artery. The hybridization signal could be observed in the ruptured aneurysms (10 cases) and unruptured ones (2 cases) often in the intima. MCP-1 mRNA appeared to be expressed by fibroblast cells in its cytoplasm. Monocyte-like cells had little cytoplasm, and the signal was seldom seen. The hybridization signal was discontinuous in the intima, MCP-1 mRNA expressed where fibroblast and monocyte-like cells assembled. One ruptured aneurysm had no signal because there were no cells only glass-like fiber. Mural thrombus showed upregulated hybridization signal in the cytoplasm of fibroblasts, phlogocytes and endotheliocytes of its micrangium.</p><p><b>CONCLUSION</b>The pathological representation of the ruptured and unruptured aneurysms and the upregulated expresion of MCP-1 in the aneurysm wall suggest that the development of aneurysm may be a course of chronic inflammation in which main inflammatory cells are monocyte-like cells.</p>

Molecular basis of the malignant phenotype of the invasive transfer of the human melanoma cell / 眼科新进展

Objective　To discuss the development and the invasive transfer mechanism.Methods　Cell culture method, p53 protenin immunohistochemistry, PCR-SSCP examination, zymography, PCM and heterogenic inouclation test inside the nude mice were adopted. Results　p53 genetic mutation existed in human melanoma cells. The fluorescent positive rate of 67KD LN-R on the surface of the melanoma cell and the average flurescent intensity are repectively WM451＞WM983A＞WM1341B＞WM35.Early WM35 did not produce MMPs; WM1341B only produced 72KDa(MMP-2),not 92KDa(MMP-9);Both WM983A at progressive stage and distal transfering tumor WM45 produced 72KDa and 92KDa. WM983A and WM451 were also seen to form evident transfering tumor inside the naked mice.Conclusion　p53 genetic mutation, the production of MMPs and the high indication of 67KD LN-R on the celluar surface are the key factors for the human melanoma cell to obtain invasive transferring abililty.