Positive Unit Research Articles

Analysis of the Pathogenesis of Gram-Negative Bacterial Sepsis in Rats Under Nano-Body

This work aimed to evaluate the effectiveness of several anti-TLR4 nanobody administration techniques in a gram-negative bacterial sepsis (GNBS) rat model. The targeting proteins for TI-Nb2 and TC-Nb6 anti-TLR4 nanobodies were TLR4203-348 and TLR4349-582, respectively. The survival times (STs) of 44 Sprague-Dawley (SD) rats were tracked in the TI-Nb2, TC-Nb6, TI-Nb2+TC-Nb6, and D0 groups (saline control). Besides, the ELISA was utilized to measure the levels of TNF-, IL-1, IL-8, and IL-10 in different groups. An automatic biochemical analyzer was employed to determine the levels of AST, ALT, AMS, CRE, and Urea. Furthermore, the rat liver and kidney tissue samples were stained with hematoxylin-eosin (HE). Cleaved-caspase-3 (CC3) protein expression (PE) in rat tissues was discovered using immunohistochemistry, and the positive unit (PU) value was computed. The TI-Nb2+TC-Nb6 group exhibited a longder ST, lower TNF-α, IL-1β, IL-8, ALT, AST, AMS, CRE, and Urea levels, and a smaller CC3 protein PU value in nucleus and cytoplasm than the TI-Nb2, TC-Nb6, and D0 groups (all P <0.05). The above findings indicated that the combined usage of TI-Nb2 and TC-Nb6 can successfully reduce the expression levels of CC3 protein, biochemical markers, and inflammatory factors. This could protect the liver, kidneys, and other organs and prolong the ST of sepsis rats.

Quantitative analysis of EZH2 expression and its correlations with lung cancer patients’ clinical pathological characteristics

Elevated Enhancer of Zeste Homologue 2 (EZH2) expression is involved in many human malignancies through epigenetically silencing related genes. However, the study of the EZH2 protein expression in lung cancer remains at the qualitative or semi-quantitative level. The present study is to elucidate the roles of EZH2 in the progression and metastasis of different subtypes of lung cancer at quantitative level. Lung carcinoma tissue microarray was constructed containing 32 normal adult lung tissues, 113 lung carcinomas and 57 lymph-node metastases. EZH2 protein expression was detected by immunohistochemistry and assessed quantitatively with Leica Q500MC image analysis system. Positive unit (PU) value was used to evaluate the protein expression intensity of positive cells from systematically selected fields under the microscope. Elevated Enhancer of Zeste Homologue 2 PU in lung carcinomas was significantly greater than that in normal lung tissues (p = 0.001). Increased EZH2 expression was correlated with histological subtypes, differentiation, TNM stage, and lymph-node metastases (p < 0.05). EZH2 PU of primary lung carcinomas was smaller than that of lymph-node metastasis (p = 0.002). EZH2 PU was not associated with patients' gender, age, smoking status, tumor location, and tumor size (p > 0.05). Elevated Enhancer of Zeste Homologue 2 PU is increased with the development of lung cancer. EZH2 may play an important role in the progression and metastasis of lung cancer.

Expression of Tiam1 in Lung Cancer and its Clinical Significance

The aim of this study was to analyze T-cell lymphoma invasion and metastasis-inducing factor 1 (Tiam1) expression in lung cancer patients. A total of 204 patients with lung cancer tissue lesions were enrolled in the present study, along with 40 cases of normal lung tissue and 40 of normal fetal lung tissue. Tiam1 protein expression level was determined using intensity quantitative analysis, for comparison in lung cancer, metastatic, normal lung, and fetal lung tissue. The positive unit (PU) of Tiam1 was 13.5 ± 5.42 in lung cancer, 5.67 ± 1.56 in normal epithelial cells, and 5.89 ± 1.45 in fetal lung epithelial cells. The value in the lung cancer tissue was significantly higher than that in the normal lung tissue and the fetal lung tissue (P<0.01). The Tiam1 PU values with lymph node metastasis and without lymph node metastasis were 15.2 ± 4.34 and 12.5 ± 4.23, respectively, and the difference was statistically significant (P<0.05). The Tiam1 PU values in different tumor, nodes, metastasis (TNM) stages, III-IV period, and I-II phase were 14.7 ± 4.14 and 11.0 ± 5.34 (P <0.05). A correlation was found between Tiam1 expression and the age of patient, tumor size, tumor type, and tumor differentiation. Tiam1 protein expression in the lung tumor tissue is significantly higher than that in the normal lung tissue and fetal lung tissue. Tiam1 expression may be closely related to lung cancer development and metastasis.

Predictive value of endometrial receptivity and pregnancy outcome by hysteroscopy examination at the phase of implantation window in unexplained infertile women

To explore predictive value of endometrial receptivity and pregnancy outcome by hysteroscopy examination at the phase of implantation window in unexplained infertile women. From Oct. 2007 to Mar. 2009, 93 unexplained infertile women underwent hysteroscopy examination at 7 approximately 9 days after a spontaneous ovulation in Family Planning Research Institute of Guangdong Province. According to the endometrial glandular openings and vascular shape, 79 cases without pathological endometrial changes were divided into 60 cases in good endometrium group and 19 cases in poor endometrium group. The following clinical parameters were analyzed and compared between two groups, including endometrial configuration, thickness, secretion, the development and number of pinopodes, vascular distribution, and the level of sex hormone, leukemia inhibitory factor (LIF) and glycodelin in the uterine flushing, and pregnancy outcome. (1) There was no statistical difference in the level of serum estrogen and progesterone at the phase of implantation window, which were (518 +/- 176) pmol/L, (40 +/- 20) nmol/L in good group and (513 +/- 244) pmol/L, (37 +/- 19) nmol/L in poor group (P < 0.05). The endometrium thickness at periovulatroy and implantation window days (1.06 +/- 0.10) cm/(1.16 +/- 0.08) cm in good group did not show significant difference with (0.93 +/- 0.12) cm/(1.02 +/- 0.10) cm in poor group (P > 0.05). The proportion of type A, B and C endometrium at periovulatory days were 63% (12/19), 37% (7/19) and 0 (0/19) in good group and 23% (14/60), 77% (46/60) and 0 (0/60) in poor group. When compared with those of type A or B between two groups respectively, it all showed statistical difference (P < 0.05). However, at phase of implantation window, endometrium configurations were all type B at both groups. (2) 90% (17/19) of women in good group and 7% (4/60) of women in poor group showed normal endometrial secretion function, which showed significant differences (P < 0.01). (3) The percentage of fully developed pinopodes and abundant pinopodes [84% (16/19) and 90% (17/19)] in good group were significantly higher than 42% (25/60) and 57% (34/60) in poor group (P < 0.05). (4) The level of CD(34) expression and microvessel density [MVD; (40.1 +/- 1.2) positive unit (PU) and (21.7 +/- 4.0)/high power field (HP)] in good group were significantly higher than (18.1 +/- 1.3) PU and (8.5 +/- 1.3)/HP in poor group (P < 0.01). (5) The level of LIF and glycodelin in uterine flushing [(72 +/- 54) ng/L and (196 +/- 20) microg/L] in good group were significantly higher than (15 +/- 16) ng/L and (116 +/- 26) microg/L in poor group (P < 0.05). (6) The rate of clinical pregnancy, spontaneous abortion and term delivery were 74% (14/19), 0 (0/14) and 100% (14/14) in good group and 23% (14/60), 14% (2/14) and 86% (12/14) in poor group, the rate of clinical pregnancy and term delivery in good group were significantly increased when compared with those in poor group (P < 0.01). Hysteroscopy examination at the phase of implantation window could reflect the development of glandular openings and vasculature. It is a preferable method to evaluate the endometrial receptivity and predict pregnancy outcome.

The relationship between transforming growth factor β1 expression and cold ischemia injury of rat donor kidney

Objective Cold ischemia injury represents an independent risk factor which favors chronic allograft nephropathy (CAN). In order to investigate the role of transforming growth factor-beta 1( (TGF-β1) in the progression of CAN, we studied the relationship between the expression of TGF-β1 and cold ischemia injury in the renal tubular epithelia of rat donor kidney. Methods A total of 24 Wistar rats were used in this study. In terms of the time of cryopreservation of donor kidney, the 24 Wistar rats were randomly divided into 3 groups: 0h group(control group), 24h group, 48h group. A block removal of donor kidney with in situ perfusion of cooling HC-A preservation solution was adopted. The rat kidney was preserved 0h, 24h and 48h at 0-4 °C respectively. The morphologic changes of proximal tubular epithelial cells in different cryopreservation time were observed under light microscope and transmission electron microscope. The expression of TGF-β1 mRNA and protein in proximal tubular epithelial cells of different cryopreservation time group were detected by in situ hybridization and immunohistochemistry analysis. Results 1. In 24h group, part of the proximal tubular epithelial cells showed slight degeneration. In 48h group, the proximal tubular epithelial cells demonstrated severe hydropic degeneration and part of the cells developed necrosis and effluxion. 2. Only a small amount of TGF-β1 protein and mRNA were expressed in the renal tubular epithelial cells of 0h group. The positive unit (PU) value of TGF-β1 protein and mRNA were 6.37 ± 2.77 and 5.29 ± 2.15, respectively. As the cold ischemia time prolonged, the PU value of TGF-β1 protein increased at 24h group (10.20 ± 3.27) and 48h group (17.17 ± 3.96) . The PU value of TGF-β1 mRNA also increased at 24h group (11.31 ± 3.34) and 48h group (19.01 ± 3.53). There was the significant difference of TGF-β1 protein PU value or mRNA PU value among these groups(P < 0.05). Conclusion There was the significant correlation between the expression of TGF-β1 and the degree of cold ischemia injury. The results suggest that TGF-β1 might play the key role in regeneration and reparation of renal tubular epithelial cell injury. The overexpression of TGF-β1 might be one of the mechanisms that initiate chronic allograft nephropathy.

Expression of p14(ARF) and E2F-1 in lung cancer in Gejiu and Xuanwei regions of Yunnan Province

To investigate the correlations between p14(ARF) and E2F-1, and the role of their alterations in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province for providing the important experiment basis in revealing the molecular mechanism and looking for new markers for early diagnosis of lung cancer. The expression of p14(ARF) and E2F-1 was detected at theirs protein level by Immunohistochemistry S-P method in 30 specimens of lung cancer of Gejiu tin miners, 30 specimens of lung cancer of Xuanwei peasants and 20 specimens of normal lung tissue. E2F-1 mRNA was detected by ISH in 25 specimens of lung cancer of Gejiu tin miners, 25 specimens of lung cancer of Xuanwei peasants and 10 specimens of normal lung tissue. The positive signals were quantitatively analysed by HPIAS-100. The positive unit (PU) of p14(ARF) and E2F-1 was 16.44 +/- 4.85 and 47.39 +/- 5.43 in Gejiu group, and 16.79 +/- 3.55 and 48.15 +/- 9.11 in Xuanwei group. Expression of p14(ARF) and E2F-1 protein in lung cancer of Gejiu and Xuanwei were statistically different compared with that in the normal lung (P < 0.01) respectively; The PU of E2F-1 mRNA was 48.58 +/- 7.75 in Gejiu group, and 49.41 +/- 8.53 in Xuanwei group, which was higher than that in normal tissue group. The differences were significant (P < 0.01). There was positive correlation between the expression of E2F-1 protein and E2F-1 mRNA in Gejiu group, Xuanwei group and normal group (P < 0.01, r = 0.833). The expression of p14(ARF) protein was significantly negatively correlated with the expression of E2F-1 protein (P < 0.01, r = -0.830). There is the over-expression of E2F-1 gene and the deletion of p14(ARF) gene in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province. Over-expression of E2F-1 protein in lung cancer may be caused by enhanced transcription.

Expression of a novel apoptosis inhibitor-survivin in colorectal carcinoma

To investigate the role of survivin expression in the pathogenesis of colorectal carcinoma. Immunohistochemistry S-P method and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were used to detect the expression of survivin and apoptotic cell in situ in colorectal cancerous tissues, para-cancerous tissues and normal tissues of 48 cases of colorectal carcinoma. The survivin positive unit (PU) was higher in cancerous tissues (38.76+/-5.14) than in para-cancerous (25.17+/-7.26) or normal tissues (0.57+/-0.03) (P<0.05). The apoptosis index (AI) of para-cancerous tissues was (7.51+/-2.63%) higher than cancerous tissues (4.65+/-1.76%). The expression of survivin was associated with pathological grade, lymph node metastasis and Dukes stage of colorectal carcinoma. Survivin expression may play an important role in carcinogenesis of colorectal carcinoma and may be associated with malignant biological behaviors of colorectal carcinoma.

Expression of Smads in lung tissue of quartz-induced pulmonary fibrosis in mice

To observe Smads protein expression in lung tissue of quartz exposed mice and to explore its association with pulmonary fibrosis in silicosis. The experimental mice were divided into control and quartz groups. 0.2 g/kg weight of quartz was injected intratracheally in quartz group. Samples were collected at the 1st, 3rd, 5th, 7th, 14th and 28th day after injection. Immunohistochemical methods with quantitative image analysis were used to assay the protein expression of transforming growth factor beta(1) (TGF-beta(1)), Smad 2/3, Smad 4, and Smad 7 protein levels. Protein expression level is presented by positive unit (PU). Smad 2/3 protein expression increased from day 3, reaching its peak level in day 14 [(42.2 +/- 2.4) PU], and decreased gradually. The elevation of Smad 4 protein level began from day 5, and the highest degree came into day 14 [(40.0 +/- 1.8) PU], decreased thereafter. The expression of Smad 7 presented a decreasing tendency at the beginning and reaching the lowest level in day 14 [(33.5 +/- 3.3) PU]. It seemed to elevate in day 28, but was still lower than the controls. There were positive correlation between Smad 2/3, Smad 4 and TGF-beta(1) (r = 0.91, r = 0.71, respectively, P < 0.05) and also between Smad 2/3 and hydroxyproline contents of lung tissue (r = 0.85, P < 0.05) except Smad 7. Smad protein may have certain association with pulmonary fibrosis in silicosis.

Quantitative study on expression of p16 multiple tumor suppressor gene in salivary gland neoplasm.

The expression of p16 gene in normal salivary acini, benign tumors and carcinomas were microscopically and quantitatively observed by using immunohistochemical method LSAB and CMIASWIN methods. The expression of p16 gene were found in all 3 groups. The positive unit (PU) was higher in tumor group and cancer group than that in normal group (P < 0.01). Furthermore, the PU of p16 was stronger in cytoplasm than in nucleus. Malignant tumors and acini surrounding the tumor revealed strong positives and weak positives respectively. The PU of p16 gene was higher in deep lobe of recurrent parotid neoplasm with incomplete capsule than that in shallow lobe of primary parotid neoplasm with complete capsule. The findings suggests that p16 gene plays an equally important role in the salivary gland tumors and tumors in other part of the body.