血细胞化学染色理论及固定问题的教与学

血细胞化学染色是近代组织化学的分支，对血细胞的各种生化成分，代谢产物，作定性，定位和定量的观察，为研究血细胞生理，病理提供科学依据。临床上亦常用来协助鉴别诊断血液病和其它疾病，并观察其疗效和预后。可见血细胞化学染色在临床医学检验中的重要性。

慢性肾炎患者外周血细胞化学ANAE变化与中医辨证关系的研究

运用细胞化学方法及显微分光光度技术对24例慢性肾炎患者和30例正常人进行外周血淋巴细胞及单核细胞的α-醋酸萘酯酶(ANAE)定性定量检测,结果正常人淋巴细胞阳性率为65.5±6.8%,其中点状颗粒型占53.6±6.7%,弥散颗粒型占11.9±3.4%,其单核细胞 ANAE 平均相对含量为54.78±3.34;肾阳虚组依次为39.7±7.4%、31.2±5.8%、8.5±2.7%及41.84±2.66;肾阴虚组依次为47.8±7.6%、42.4±6.3%、5.4±2.2%和51.26±2.59。经统计学处理,肾炎组与正常人、肾阴虚与肾阳虚之间差异均有不同程度的显著性意义,经中医辨证治疗后,各项指标均有一定程度的改善。并就慢性肾炎的中医辨证与 T 细胞亚群的关系作了讨论。

《血细胞化学染色》视听教材的制作及应用效果

血细胞化学染色是实验教学中的重点和难点,本课题组应用现代教育技术编制了《血细胞化学染色》视听教材,并应用到实验教学中,促进了实验教学形式的变革,增强了学生学习效果,提高了血细胞化学染色的教学质量。

全血白细胞吞噬功能测定的改进及临床应用

目的建立一个同时测定多个样品的全血白细胞吞噬功能测定的方法,缩短临床标本检测的时间。方法在全血白细胞吞噬功能测定实验中,通过编程和进样的改进,实现多个循环的连续测定的时间优化。增加一个分配器为进样器,满足大体积一次进样的要求。结果改进后的方法与原方法的对比实验相关性很好(R2=0.99),而效率提高了24倍。21例肺癌患者手术治疗恢复后,其CL的CL60min,和CPM和Pt强度明显增加(P<0.05)。结论该文探讨的改进方法是可行的,使全血CL测定可能成为肺癌患者白细胞活性功能及观察疗效的一个指标,增强了全血白细胞吞噬功能检测的临床实用性。

INHIBITION OF FARNESYL PROTEIN TRANSFERASE AND P21^(RAS) MEMEBRANE ASSOCIATION BY D-LIMONENE IN HUMAN PANCREAS TUMOR CELLS IN VITRO

The monoterpene d limonene inhibit the plasma membrane associated P21 ras expression and the posttranslational isoprenylation of P21 ras , a mechanism that may contribute to its efficacy in the chemoprevention and therapy of chemically induced rodent cancers and some human solid tumor cells. In the present study,the relative abilities of d limonene to inhibit membrane associated P21 ras expression in pancreas tumor cell(PaCa) was carried out with Western blotting, and the inhibition of farnesyl protein transferase (FTPase) activity during the Ras protein isoprenylation and cell proliferation were determined.Concomitantly,the effects of d limonene on P21 ras localization by immunohistochemistry and H ras oncogene expression in PaCa tumor cell line by Northern blotting were observed. The results showed that d limonene inhibited FPTase activity, thus to reduce P21H ras isoprenylation. d limonene could decrease P21 ras membrane association and increase cytosolic accumulation of P21 ras . This phenomenon was also noted when d limonene treated PaCa cells were stained immunohistochemically with anti P21 ras antibody. It is suggested that the inhibition of FPTase activity was closely related with the inhibiton of P21 ras membrane association and the alteration of P21 ras localization. Inhibition of farnesylation of P21 ras altered their intracellular localization and, hence, disrupted their biological activity,but no relationship with H ras oncogene expression was found.

Genetic Correction and Hepatic Differentiation of Hemophilia B-specific Human Induced Pluripotent Stem Cells

Objective To genetically correct a disease-causing point mutation in human induced pluripotent stem cells （iPSCs） derived from a hemophilia B patient. Methods First, the disease-causing mutation was detected by sequencing the encoding area of human coagulation factor IX （F IX） gene. Genomic DNA was extracted from the iPSCs, and the primers were designed to amplify the eight exons of F IX. Next, the point mutation in those iPSCs was genetically corrected using CRISPR/Cas9 technology in the presence of a 129-nucleotide homologous repair template that contained two synonymous mutations. Then, top 8 potential off-target sites were subsequently analyzed using Sanger sequencing. Finally, the corrected clones were differentiated into hepatocyte-like cells, and the secretion of F IX was validated by immunocytochemistry and ELISA assay.Results The cell line bore a missense mutation in the 6th coding exon （c.676 C〉T） of F IX gene. Correction of the point mutation was achieved via CRISPR/Cas9 technology in situ with a high efficacy at about 22% （10/45） and no off-target effects detected in the corrected iPSC clones. F IX secretion, which was further visualized by immunocytochemistry and quantified by ELISA in vitro, reached about 6 ng/ml on day 21 of differentiation procedure. Conclusions Mutations in human disease-specific iPSCs could be precisely corrected by CRISPR/Cas9 technology, and corrected cells still maintained hepatic differentiation capability. Our findings might throw a light on iPSC-based personalized therapies in the clinical application, especially for hemophilia B.

Ankaferd hemostat in the management of gastrointestinal hemorrhages

Gastrointestinal (GI) bleeding refers to any hemorrhage ascribed to the pathologies of the gastrointestinal tract,extending from the mouth to the anal canal.Despite the recent improvements in the endoscopic,hemostatic and adjuvant pharmacologic techniques,the reported mortality is still around 5%-10% for peptic ulcer bleeding and about 15%-20% for variceal hemorrhages.Although endoscopic management reduces the rates of re-bleeding,surgery,and mortality in active bleeding;early recurrence ratios still occur in around 20% of the cases even with effective initial hemostatic measures.In this quest for an alternative pro-hemostatic agent for the management of GI bleedings,Ankaferd blood stopper (ABS) offers a successful candidate,specifically for "difficult-to-manage" situations as evidenced by data presented in several studies.ABS is a standardized mixture of the plants Thymus vulgaris,Glycyrrhiza glabra,Vitis vinifera,Alpinia officinarum,and Urtica dioica.It is effective in both bleeding individuals with normal hemostatic parameters and in patients with deficient primary and/or secondary hemostasis.ABS also modulates the cellular apoptotic responses to hemorrhagic stress,as well as hemostatic hemodynamic activity.Through its effects on the endothelium,blood cells,angiogenesis,cellular proliferation,vascular dynamics,and wound healing,ABS is now becoming an effective alternative hemostatic medicine for gastrointestinal bleedings that are resistant to conventional anti-hemorrhagic measurements.The aim of this review is to outline current literature experience suggesting the place of ABS in the management of GI bleeding,and potential future controlled trials in this complicated field.

Deformation and Motion of a Red Blood Cell in a Shear Flow Simulated by a Lattice Boltzmann Model

A lattice Boltzmann model is presented to simulate the deformation and motions of a red blood cell （RBC） in a shear flow. The curvatures of the membrane of a static RBC with different chemical potentiM drops calculated by our model agree with those computed by a shooting method very well. Our simulation results show that in a shear flow, biconcave RBC becomes highly flattened and undergoes tank-treading motion. With intrinsically parallel dynamics, this lattice Boltzmann method is expected to find wide applications to both single and multi-vesicles suspension as well as complex open membranes in various fluid flows for a wide range of Reynolds numbers.

Expression of caveolin-1 in clear cell renal cell carcinoma and its correlation with microvessel density

Objective:The aim of the study was to investigate the clinicopathologic significance of caveolin-1 expression in clear cell renal cell carcinoma (CCRCC) and its correlation with microvessel density (MVD).Methods:The expression of caveolin-1 was detected by the immunohistochemistry method,while the microvessel density was detected by the immunohistochemistry expression of CD34.Results:In the CCRCCs,the positive rate of caveolin-1 was 67.4%,the over expression of caveolin-1 was not related with sex and age,but related with clinicopathologic parameter,such as tumor sizes,clinical TMN stage,nuclear stage and survival time (P < 0.05).The MVD of positive caveolin-1 cases was significantly higher than that without caveolin-1 expression (P < 0.05).Conclusion:The expression of caveolin-1 is helpful in the prognostic evaluation of CCRCCs and it may be involved in the tumor angiogenesis.

The feasibility of antitumor drugs chemosensitivity testing by flow cytometry

Objective: To investigate the feasibility of chemosensitivity testing of antitumor drugs by flow cytometry in clinical applications so as to provide experimental and theoretical basis for the establishment of a novel antitumor drugs sensitivity testing and the screening of particular antitumor drugs. Methods: Detect the apoptosis rate of 12 cases of Molt-4 cell line, 57 cases of fresh clinical gastrointestinal tumor cells by Sub-G1 and Annexin V assay of flow cytometry under the effects of antitumor drugs at different times and the outcomes were compared with the ones of the MTT (3-(4,5-dimethylthiazolyl-2) -2,5-diphenyltetrazolium bromide) assay. Results: The lethality of drugs on Molt-4 cell, clinical gastrointestinal tumor cells had a positive correlation with the acting time of antidrugs by employing Annexin V, Sub-G1 and MTT assay. Drug-incurring maximum lethality of Annexin V assay was higher than MTT colorimetric assay, that of Sub-G1 was lower than MTT assay, the virtual times of Annexin V and Sub-G1 assay were obviously earlier than that of MTT colorimetric assay. Conclusion: Annexin V and Sub-G1 assay of flow cytometry can be taken as potent protocols testing anti-tumor drug chemosensitivity. Annexin V assay is featured by more sensitive, concise, reliable compared with the classical chemosensitivity testing assay of MTT colorimetric assay and it possesses clinical applied value.

Constructed Model of Cost/Benefit Analysis Strategy for Stem Corn Borer Sesamia cretica

An integrated pest management model of Cost/Benefit analysis strategy was constructed for stem corn borer Sesamia cretica using complementary control measures of different resistant genotypes of corn with the chemical insecticide diazinon 60%. Based on Cost/Benefit analysis result, the resistant genotype （SAKHA 9433） provides maximum economic value of production at the model point where no spray of insecticide is applied. The applications of one or two sprays do not justify the use of chemical insecticide but rather result in economic loss since the reduction in borer damage value, due to diazinon use, is lower than the cost of control （spray）. However, the applications of one spray for the moderate resistant genotype （IPA 2052） and two sprays for the sensitive genotype （CML 323） during corn growing season would be of value to cover the cost of control （spray） but do not achieve similar economic value of revenue comparing with the resistant genotype.

AGING OF HUMAN MATURE ERYTHROCYTES IS LIKE A PROCESS OF APOPTOSIS IN ENUCLEATED CELL

Apoptosis of nucleated cells is well known, but how about the unnucleated cells is still not elucidated. In the present paper, the morphological and biochemical features of the aged erythrocytes were observed and compared with the characteristic events of apoptosis. Membrane of aged erythrocytes tends to shrink, protrude, from vesicle and lose lipid asymmetry. Aged erythrocytes were removed by phagocytosis. Both of the events are very similar to the apoptotic nucleated cells. The authors suggested that aging of erythro- cytes is also a process of apoptosis.

Combining proteomics, serum biomarkers and bioinformatics to discriminate between esophageal squamous cell carcinoma and pre-cancerous lesion

Objective: Biomarker assay is a noninvasive method for the early detection of esophageal squamous cellcarcinoma (ESCC). Searching for new biomarkers with high specificity and sensitivity is very important for the earlydetection of ESCC. Serum surface-enhanced laser desorption/ionization-time of flight mass spectrometry(SELDI-TOF-MS) is a high throughput technology for identifying cancer biomarkers using drops of sera. Methods: Inthis study, 185 serum samples were taken from ESCC patients in a high incidence area and screened by SELDI. Asupport vector machine (SVM) algorithm was adopted to analyze the samples. Results: The SVM patterns success-fully distinguished ESCC from pre-cancerous lesions (PCLs). Also, types of PCL, including dysplasia (DYS) and basalcell hyperplasia (BCH), and healthy controls (HC) were distinguished with an accuracy of 95.2% (DYS), 96.6% (BCH),and 93.8% (HC), respectively. A marker of 25.1 kDa was identified in the ESCC patterns whose peak intensity wasobserved to increase significantly during the development of esophageal carcinogenesis, and to decrease obviously after surgery. Conclusions: We selected five ESCC biomarkers to form a diagnostic pattern which can discriminateamong the different stages of esophageal carcinogenesis. This pattern can significantly improve the detection ofESCC.