一种简便的宫颈细胞学染色法

子宫颈癌是女性最常见的恶性肿瘤,尤其在经济落后及偏远地区,宫颈癌的发病率占女性肿瘤的较大比例.随着子宫颈癌预防普查工作的广泛开展,使许多无症状的子宫颈原位癌和早期癌能及时发现,提高了宫颈癌的五年治愈率.宫颈刮片检查是发现宫颈癌的最有效方法之一,目前常用苏木素及伊红染色(HE)后进行细胞学检察.近来,我们改用美蓝(MB)染色观察了1 800张宫颈刮片,现将结果报道如下.

液基细胞学制片巴氏染色临床诊断真菌性阴道炎的价值

目的探讨液基细胞学制片巴氏染色诊断真菌性阴道炎的临床价值。方法选取2020年4月至2021年6月期间在河南中医药大学第三附属医院门诊或住院的具有典型临床症状、初步诊断为真菌性阴道炎的251例患者进行研究。共采集到有效标本230例,阴道微生态方法检测128例(有效标本122例),白带常规检测123例(有效标本108例),所有标本均接受液基细胞学制片巴氏染色方法检测,以真菌培养结果作为诊断金标准,绘制2×2配对四格表计算相关诊断学指标,绘制受试者工作特征(ROC)曲线计算曲线下面积(AUC)分析液基细胞学制片巴氏染色用于诊断真菌性阴道炎的价值。结果采用阴道微生态、液基细胞学两种方法检查的122例患者,经真菌培养共计检测出真菌感染患者52例,其中最主要的真菌类型为白色念珠球菌25例(48.08%),其次为光滑念珠球菌18例(34.62%);阴道微生态诊断阴道真菌感染的灵敏度为75.00%、特异度为91.43%、阳性预测值为86.67%、阴性预测值为83.12%、ROC曲线下面积AUC值为0.784;液基细胞学诊断阴道真菌感染的灵敏度为95.74%、特异度为90.67%、阳性预测值为86.53%、阴性预测值为97.14%、ROC曲线下面积AUC值为0.918;接受白带常规、液基细胞学两种方法检查的108例患者,经真菌培养共计检测出真菌感染患者51例,其中最主要的真菌类型为白色念珠球菌27例(51.92%),其次为光滑念珠球菌15例(28.85%);白带常规诊断阴道真菌感染的灵敏度为96.08%、特异度为75.44%、阳性预测值为77.78%、阴性预测值为95.56%、ROC曲线下面积AUC值为0.824;液基细胞学诊断阴道真菌感染的灵敏度为96.97%、特异度为88.71%、阳性预测值为86.27%、阴性预测值为96.49%、ROC曲线下面积AUC值为0.914。结论液基细胞学制片巴氏染色作为一种新的真菌定性检测方法,应用于临床诊断真菌性阴道炎较阴道微生态方法具有更高的灵敏度,较白带常规具有更高的特异性,同时保证诊断的准确性,具有一定的临床应用价值。

良好甲状腺结节细针穿刺细胞学HE染色探究

随着甲状腺结节性病变发病率逐渐升高,甲状腺结节细针穿刺(FNA)技术广泛应用于临床。通过对甲状腺结节FNA传统细胞涂片及液基细胞学HE染色回顾性分析,摸索总结针对甲状腺结节FNA细胞学HE染色最佳条件,提高染色质量,为病理医师准确筛查甲状腺结节性质,顺利开展后续工作提供保障。作者在日常中不断摸索、改进、总结经验,现将良好甲状腺结节FNA细胞学HE染色方法及影响染色质量的原因介绍,分析如下。

探讨毛细式液基细胞学薄层染色技术对宫颈癌筛查应用价值研究

目的探讨毛细式液基细胞学薄层染色技术(TCT)对宫颈癌筛查的临床应用价值。方法随机选取2011年6月—2013年7月期间在该院行宫颈癌筛查的患者480例,所有患者均行TCT检查,将其中意义不明确的不典型鳞状上皮细胞(ASCUS)以上病变视为阳性病变,并对其进行阴道镜活检,将活检结果和细胞学检测结果进行对比分析。结果 480例患者中,共计得到102例阳性标本,阳性率为21.25%;其中35例异常者的TCT检查的同阴道镜活检的符合率为88.57%。结论在宫颈癌筛查中应用毛细式液基细胞学薄层染色检查,可有效提高筛查质量,对于宫颈癌的预防和治疗具有十分重要的意义。

骨髓细胞学联合骨髓铁染色、血清铁蛋白在不明原因贫血患者诊断中的意义研究

目的探讨骨髓细胞学联合骨髓铁染色、血清铁蛋白在不明原因贫血患者诊断中的意义。方法回顾性分析205例不明原因贫血患者临床资料,入院行骨髓细胞学检查,并对骨髓涂片进行铁染色,在低倍光镜及油镜下观察细胞内、外铁,并计算细胞内铁阳性率;利用化学发光仪对血清铁蛋白(SF)含量检测。结果205例贫血患者中检出缺铁性贫血、巨幼细胞性贫血、慢性病性贫血、溶血性贫血、急性白血病、再生障碍性贫血,其中缺铁性贫血患者占比最高40%,另有29例贫血原因不能明确;血清铁蛋白与骨髓细胞内铁及外铁均存在正相关;将骨髓细胞学联合骨髓铁染色、血清铁蛋白三者联合检查鉴别诊断不明原因贫血患者的灵敏度、特异度以及准确率最高。结论骨髓细胞学联合骨髓铁染色、血清铁蛋白在不明原因贫血患者诊断中起到关键作用,可准确诊断和鉴别大部分不明原因贫血,为后续治疗提供重要依据。

Histological Structure Difference of Dog's Olfactory Bulb Between Different Age and Sex

The purpose of this article is to detect sex and age difference in the structure of the olfactory bulb in dogs by histological methods. The thickness of the olfactory bulbs layers and its main cells were analyzed comparatively with the methods of HE-staining and statistics, through which we studied the development course of dogs＇ olfactory bulb and the structural differences which affect the olfaction in both males and females. The results showed that between both male and female juveniles and adult males and females, the difference in thickness of each layer is not significant. But the difference in quantity of mitral cells between adult males and females was significant. Meanwhile, the structure of every layer in juvenile dogs was apparent while the volume and the weight of adult dogs＇ olfactory bulb and each layer＇s width increased significantly. On the other hand, the density of each layer＇s cells decreased apparently. Our results demonstrated that the olfactory bulb developed with age, and the apparent differences in morphology and quantity of mitral cells between males and females may be one of the reasons leading to the sexual variations of olfactory sensitivity.

Characterization of Two Human Lung Adenocarcinoma Cell Lines by Reciprocal Chromosome Painting

Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines： A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified An A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.

Genetics of hepatocellular carcinoma

The completely assembled human genome has made it possible for modern medicine to step into an era rich in genetic information and high-throughput genomic analysis. These novel and readily available genetic resources and analytical tools may be the key to unravel the molecular basis of hepatocellular carcinoma (HCC). Moreover, since an efficient treatment for this disease is lacking, further understanding of the genetic background of HCC will be crucial in order to develop new therapies aimed at selected targets. We report on the current status and recent developments in HCC genetics. Special emphasis is given to the genetics and regulation of major signalling pathways involved in HCC such as p53, Wnt- signalling, TGFβ, Ras, and Rb pathways. Furthermore, we describe the influence of chromosomal aberrations as well as of DNA methylation. Finally, we report on the rapidly developing field of genomic expression profiling in HCC, mainly by microarray analysis.

Nuclear β-catenin expression as a prognostic factor in advanced colorectal carcinoma

AIM： To investigate the changing pattern of β-catenin expression and its prognostic value in advanced colorectal cancer （CRC）. METHODS： Archival tumor samples were analyzed for β-catenin using immunohistochemistry （IHC） in 95 patients with advanced CRC. RESULTS： Membranous β-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 （58%） cases showed nuclear expression. In univariate （Kaplan-Meier） survival analysis, only the nuclear index （NI） was a significant predictor of disease free survival （DFS） （P = 0.023; n = 35）, with a NI above the median associated with longer DFS （34.2 too） than those with a NI below the median （15.5 too） （P = 0.045, ANOVA）. The other indices were not significant predictors of DFS, and none of the three tested indices （for membranous, cytoplasmic, or nuclear expression） predicted diseasespecific survival （DSS）. However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS （P = 0.041） and DSS （P = 0.046）. CONCLUSION： Nuclear β-catenin expression provides additional information in predicting patient outcome in advanced CRC.

DIFFERENCE IN BIOLOGICAL CHARACTERISTICS AND SENSITIVITY TO CHEMOTHERAPY AND RADIOTHERAPY BETWEEN INTRAHEPATIC AND EXTRAHEPATIC CHOLANGIOCARCINOMA CELLS IN VITRO

Objective To investigate and compare the biological characteristics and sensitivity to chemotherapy and radiotherapy of intrahepatic and extrahepatic cholangiocarcinoma cells in vitro.Methods The intrahepatic and extrahepatic cholangiocarcinoma cell lines were established,and cells with steady passage were chosen to study the biological characteristics including morphology,growth dynamics,chromosome,and levels of cancer antigen(CA)125,CA19-9,alpha-fetoprotein(AFP),and carcino-embryonic antigen(CEA).Meanwhile,MTT assay was used to determine the sensitivity of both kinds of cells to 6 chemotherapeutic drugs,including cisplatin,paclitaxel,harringtonine,5-fluorouracil,vincristine,and aclacimomycin,and the inhibitory rate of cells under the irradiation of 10 Gy ray was also measured.Results The intrahepatic cholangiocarcinoma cells were mostly fusiform in shape,and extrahepatic cholangiocarcinoma cells were mostly round or polygon in shape.Their doubling time was 26.3 hours and 23.1 hours,respectively.Their average number of chromosomes was 59(range,38-84)and 67(range,49-103),respectively.The chromosome karyotypes of most intrahepatic cholangiocarcinoma cells were hyperdiploid and hypotriploid,while hypertriploid was predominant in extrahepatic cholangiocarcinoma cells.The level of CA 125 in supernatant of extrahepatic cholangiocarcinoma cells increased obviously,while levels of other determined tumor markers in both kinds of cells were all within normal range.The intrahepatic cholangiocarcinoma cells were low sensitive to cisplatin and paclitaxel,but not sensitive to the other 4 chemotherapeutic drugs.The extrahepatic cholangiocarcinoma cells were high sensitive to cisplatin,but not sensitive to the other 5 drugs.Both kinds of cells had poor sensitivity to radiotherapy.Conclusions Intrahepatic and extrahepatic cholangiocarcinoma cells show differences in shape,doubling time,chromosome karyotype,tumor marker level,and chemosensitivity,whereas they both have poor radiosensitivity.Though they are similar in histopathology,they have different growth characteristics and have discrepancy in treatment and prognosis.

Upregulated CD133 expression in tumorigenesis of colon cancer cells

AIM: To analyze the upregulated CD133 expression in tumorigenesis of primary colon cancer cells. METHODS: Upregulated CD133 expression in tumorigenesis of colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) was analyzed by flow cytometry. Human colon cancer tissue samples were stained with anti-human CD133. SW620 cells were sorted according to the CD133 expression level measured by fluorescence-activated cell sorting. Spheroids of colorectal cancer cells were cultured with the hanging drop. Expression of CD133 and Lgr5 in spheroids of colorectal cancer cells and monolayer culture was detected by RT-qPCR. Spheroids of colorectal cancer cells were analyzed using anti-human CD133 with immunohistochemical staining. RESULTS: CD133 antigen was expressed in colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) as well as in primary and metastatic human colon cancer tissues. However, the CD133 was differently expressed in these cell lines and tissues. The expression levels of CD133 and Lgr5 were significantly higher in spheroids of parental, CD133hi and CD133-cells than in their monolayer culture at the mRNA level (P < 0.05). Immunohistochemical staining of spheroids of CD133-cells showed that CD133 was highly expressed in colorectal cancer cell lines. CONCLUSION: Upregulated CD133 expression plays a role in tumorigenesis colorectal cancer cells, which may promote the expression of other critical genes that can drive tumorigenesis.

Expression of phosphatase and tensin homolog deleted on chromosome ten in liver of athymic mice with hepatocellular carcinoma and the effect of Fuzheng Jiedu Decoction

AIM： To explore the expression of phosphatase and tensin homolog deleted on chromosome ten （PTEN） in liver of athymic mice with hepatocellular carcinoma （HCC） and the effect of Fuzheng Jiedu Decoction （FJD）. METHODS： Forty eight male BALB/c athymic mice models were built by Bel-7402 with an indirect method. After 24 h of postoperation, the 48 athymic mice were distributed randomly into 4 groups： A, B, C, D, each group had 12 athymic mice. Group A were were treated by intragastric administration with FT207 （Tegafur） for 4 wk. Group B, C and D were treated by intragastric administration with FJD （complex prescription of Chinese crude drug） that had been delegated into 3 kinds of density as the low, middle, and high for 4 wk. At last, athymic mice were put to death, live time, volume of tumors, exponent of tumors and the tumor metastasis in livers were observed; and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry. RESULTS： Four weeks later, the total survival rate in treatment group （A ＋ B ＋ C） was 50% and higher than the control group （0%） treated by FT207, （P 〈 0.01）. The survival rate in group A, B, C was higher than in group D, and except group A with D, there was significant differentces （Fisher＇s Exact Test P = 0.05 or 0.01）. And no differences were observed between the treatment groups and the control group in volume of tumors and exponent of tumors （P 〉 0.05）. Tumor metastasis in livers of the treatment group was less than the controls （Fisher＇s Exact Test, P = 0.021）. The result of immunohistochemistry showed that the intensity of PTEN in latero-cancer tissue was the highest, and then the hepatic tissue, the lowest was cancer tissue （Kruskal- Wallis test, X^2 = 60.67, P = 0.000）. It also showed that the intensity of PTEN in treatment groups （A, B, C） was higher than the control group （D） （F = 5.90, P = 0.002 in hepatic tissue and F = 15.99, P = 0.000 in latero-cancer tissue and X^2 = 26.08, P = 0.000 in cancer tissue）, and group B is the highest in the treatment groups （P 〈 0.05, r = 0.01. respectively）. However, there was no significant statistic difference between group A and group C （P 〉 0.05）. CONCLUSION： FJD can prolong the survival time and decrease tumor metastasis in livers of these experimental mice. Mechanisms of FJD healing HCC may partially be explained by enhancing the expression of PTEN in liver.

FSCE用于乳腺肿瘤术中诊断的研究

目的：探讨快速染色刮取涂片细胞学检查用于乳腺肿瘤术中快速诊断。方法：采用FSCE对90例乳腺肿瘤术中送检快速检查标本进行刮取涂片细胞学诊断，再与快速病理诊断对照，看其是否相吻合，最后与常规病理检查相比较，看其准确率。结果：FSCE制片全过程不超过1min，准确率98．7％，其中60例恶性肿瘤无一例假阳性，29例良性肿瘤无一例假阴性，1例未确诊者为粘液腺癌。结论：该方法用于乳腺肿瘤术中快速诊断，其时间快，设备简单，操作方便，准确率高，未见假阳性和假阴性。

Clinical cytogenetics and molecular cytogenetics

The short report will be focused on helping our students to understand commonly used conventional and cutting edge cytogenetic techniques and their clinical applications, the advances and drawbacks of each technique, and how to pick the right test(s) for a specific patient in order to achieve a proper diagnosis efficiently and economically.

Enhanced apoptosis during early neuronal differentiation in mouse ES cells with autosomal imbalance

Although particular chromosomal syndromes are phenotypicaUy and clinically distinct, the majority of individuals with autosomal imbalance, such as aneuploidy, manifest mental retardation. A common abnormal phenotype of Down syndrome （DS）, the most prevalent autosomal aneuploidy, shows a reduction in both the number and the density of neurons in the brain. As a DS model, we have recently created chimeric mice from ES cells containing a single human chromosome 21. The mice mimicked the characteristic phenotypic features of DS, and ES cells showed a higher incidence of apoptosis during early neuronal differentiation in vitro. In this study, we examined the induction of anomalous early neural development by aneuploidy in mouse ES cells by transferring various human chromosomes or additional mouse chromosomes. Results showed an elevated incidence of apoptosis in all autosome-aneuploid clones examined during early neuronal differentiation in vitro. Further, cDNA microarray analysis revealed a common cluster of down-regulated genes, of which eight known genes are related to cell proliferation, neurite outgrowth and differentiation. Importantly, targeting of these genes by siRNA knockdown in normal mouse ES cells led to enhanced apoptosis during early neuronal differentiation. These findings strongly suggest that autosomal imbalance is associated with general neuronal loss through a common molecular mechanism for apoptosis.

In vitro cultivation and differentiation of fetal liver stem cells from mice

During embryonic development, pluripotent endoderm tissue in the developing foregut may adopt pancreatic fate or hepatic fate depending on the activation of key developmental regulators. Transdifferentiation occurs between hepato- cytes and pancreatic cells under specific conditions. Hepatocytes and pancreatic cells have the common endodermal progenitor cells. In this study we isolated hepatic stem/progenitor cells from embryonic day (ED) 12-14 Kun-Ming mice with fluorescence-activated cell sorting (FACS). The cells were cultured under specific conditions. The cultured cells deploy dithizone staining and immunocytochemical staining at the 15th, 30th and 40th day after isolation. The results indicated the presence of insulin-producing cells. When the insulin-producing cells were transplanted into alloxan- induced diabetic mice, the nonfasting blood glucose level was reduced. These results suggested that fetal liver stem/ progenitor cells could be converted into insulin-producing cells under specific culture conditions. Fetal liver stem/ progenitor cells could become the potential source of insulin-producing cells for successful cell transplantation therapy strategies of diabetes.

Extramedullary plasmocytoma associated with a massive deposit of amyloid in the duodenum

We report a rare case of extramedullary plasmocytoma associated with a massive deposit of amyloid in the duodenum. A 72-year-old Japanese man was admitted to our hospital presenting with a 3-mo history of epigastric pain, vomiting and weight loss. On computed tomography （CT） a wall thickening of the fourth part of the duodenum was observed. Multiple biopsies obtained from the lesion showed infiltration of plasma cells and lymphocytes, but they were not conclusive. The patient underwent resection of the lesion and, on histopathological examination, the lesion consisted of a dense and diffuse infiltrate of plasma cells and a few admixed lymphocytes with reactive follicles extending to the muscular propria. An extensive deposition of amyloid was also observed. Immunohistochemical stains revealed that a few plasmacytoid cells showed λ light chain staining, though most were κ： light chain positive. These cells also were positive for CD138 and CD56 but negative for CD20 and CD79. The findings were consistent with extramedullary plasmocytoma associated with a massive deposit of amyloid in duodenum. A subsequent workup for multiple myeloma was completely negative. The patient showed no signs of local recurrence or dissemination of the disease after 12 mo follow-up. Because of the association of plasmocytoma and amyloidosis, the patient must be followed up because of the possible systemic involvement of the neoplasm and amyloidosis in future.

Polytene chromosomes of an Indian Himalayan black fly Simulium (Nevermannia) praelargum (Diptera:Simuliidae)

High quality polytene chromosome maps （n=3） of a Himalayan Simuliid Simulium praelargum Datta, 1973 are presented and represent the first cytological description of a taxon found in the feuerborni group, subgenus Nevermannia. Polytene chromosomes one （I） and two （II） are metacentric, chromosome three （III） is submetacentric with the length of each chromosome occupying 37.25 %, 31.36 % and 31.34 % of the total complement length, respectively. Typical simuliid diagnostic intergeneric chromosomal markers are found within the polytene complement of this species. The nucleolar organizer （N.O.） is found at the base of the short arm of chromosome one （IS）, the Ring of Balbiani （R.B.）, double bubble （D.B.） and triad occur in the short arm of chromosome two （IIS）, the Parabalbiani Ring （EB.） and grey band （gb） occur in the long arm of chromosome two （IIL） and the Blister （BL） and Capsule （Ca） occur in the short arm of chromosome three （IIIS）.Terminal bands at the end of IIIS are heterochromatinized and present atypically with respect to other simuliid fauna. Populations studied so far are unique among the Simuliidae in that they exhibit chromosome structural monomorphism. These high resolution polytene chromosome maps will form the basis for future cytological characterization and phylogenetic comparisons amongst members of the feuerborni group

Cytogenetic and Clinical Analysis of 340 Chinese Patients with Primary Amenorrhea

Objective To analyze the relationship between karyotypes and clinic features of patients with primary amenorrhea.Methods G banding was done for 340 patients with primary amenorrhea to facilitate individual chromosome identification,and if specific staining for certain portions of the chromosome was necessary,C banding was used.The clinical data were recorded by physical examination and ultrasound scanning.Results Karyotype analysis of the 340 patients revealed that 180(52.94%) patients had normal female karyotypes and 160(47.06%) patients had abnormal karyotypes.The abnormal karyotypes included abnormal X chromosome(150 patients),mosaic X-Y chromosome(4 patients),abnormal autosome(5 patients),and X-autosome translocation(1 patient).The main clinical manifestations in patients with primary amenorrhea were primordial or absent uterus(95.9%),invisible secondary sex features(68.8%),little or absent ovary(62.6%),and short stature(30.0%).The incidence of short stature in patients with X chromosome aberration(46%,69/150) was significangly higher that in patients with 46,XX(9.44%,17/180) as well as 46,XY(6.67%,3/45;χ2=146.25,P=0.000).All primary amenorrhea patients with deletion or break-point at Xp11.1-11.4 were short statures.Conclusions One of the main reasons of primary amenorrhea is choromosome abnormality,especially heterosome abnormality.It implies the need to routinely screen chromosomal anomalies for such patients.There might be relationship between Xp11.1-11.4 integrity and height improvement.

Granular cell tumor of the urinary bladder: A case report

We reported a case of a GCT of the urinary bladder and review the literature.A 23-year-old female presented with dysuria that had lasted for the previous 6 months.MRI revealed a 3×2.5 cm global mass in the anterior wall of urinary bladder.Cystoscopy showed a semispherical tumor approximately 3 cm in diameter that was covered with normal bladder mucosa and extended from the bladder neck to the anterior wall of the bladder.The patient underwent transurethral resection of the tumor.Histological examination and immunohistochemical staining showed a granular cell tumor(GCT).There were no features suggesting a malignant phenotype.On 6 months follow-up,the patient has remained free of bladder recurrence.We recommend careful pathologic assessment for establishing the appropriate diagnosis and either a conservative or aggressive surgical treatment for benign or localized malignant GCT of the urinary bladder,respectively.