Analysis on Factors Influencing Posttransfusion Effectiveness in Plasma Transfusion of 4423 Cases-Times / 中国实验血液学杂志

OBJECTIVE@#To investigate the related factors influencing plasma transfusion efficacy so as to improve the plasma transfusion efficiency.@*METHODS@#According to the clinical symptoms and the laboratorial results, the patients were divided into transfusion efficient and inefficient groups. A total of13090.8 units of plasma were transfused to 4423 patients. The clinical symptoms and the hemorrhage related index per- and pro-transfusion, plasma components sorts, storage time, and the dose of plasma (kg/ml) transfusion were analyzed.@*RESULTS@#The largest transfusion volume of plasma were in intensive care unit (ICU) accounted for 30.36%, the largest blood plasma per patient transfusion was in cardiac surgery (3.96 U). The analysis of transfusion efficiency showed that in terms of patient age, there were difference in transfusion efficiency among the patients with different ages (P＜0.001). The effective transfusion rate in the group of age ＜18 was 53%, which was higher than that in group of age 18-60(41%) and group of age ＞60 (30%); in terms of sex, the effective transfusion rate in female group was higher than that in male group (42% vs 37%) (P＜0.001); in terms of transfusion plasma volume/body weight, there were differences in transfusion efficiency (P＞0.05). The multi-factor logistic regression analysis showed that there was no significant correlation among the plasma sorts, storage time of the plasma pre-transfusion and transfusion efficiency(P＞0.05). The analysis of the non-hemolytic fever reaction caused by plasma transfusion revealed that there was no statistical difference between the plasma and the leukocyte-depleted plasma groups (P＞0.05).@*CONCLUSION@#The plasma transfusion effectiveness relates with age and sex, but not relates with the transfusion plasma voume/body weight, plasma sorts, and the duration of storage.

Effects of propofol sedation on BDNF-TrkB/p75 signal and cognitive function in rat hippocampus / 上海交通大学学报（医学版）

Objective·To detect the effects of propofol sedation on cognitive function in rats and its mechanism. Methods?·?Forty-eight SD rats were randomly divided into three groups, i.e. control group, 100?mg/kg group and 300?mg/kg group. Rats were administrated intraperitoneally with propofol (10?mg/mL, 100?mg/kg or 300?mg/kg). The mRNA levels of brain derived neurotropic factor (BDNF)-TrkB/p75 signal molecules in rat hippocampus were evaluated by realtime PCR 45 min after propofol treatment. Learning and memory ability was examined by inhibitory avoidance (IA) test after propofol treatment. Results?·?The mRNA levels of BDNF in the hippocampal tissue were (1.20±0.13) fold (P=0.002) and (88±12) % (P=0.044) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group after injection of propofol. The mRNA levels of TrkB were (1.01±0.11) fold ( P=0.982) and (86±11) % (P=0.018) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group. The mRNA levels of p75 were (1.02±0.10) fold (P=0.778) and (1.59±0.18) fold (P=0.000) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group. There was no significant difference of the 24 h IA memory retention latency between 100?mg/kg group and control group. The 24 h IA memory retention latency in 300?mg/kg group was significantly decreased compared with control group (P=0.028) and 100?mg/kg group (P=0.020). Conclusion?·?Propofol dose-dependently regulates the expression of BDNF-TrkB/p75 signal molecules, and high dose propofol may reduce cognitive function via BDNF-TrkB/p75 signal.

Propofol activates rat hippocampal astrocytes time-dependently via ERK signaling pathway / 上海交通大学学报（医学版）

Objective·To detect the effects of propofol on rat hippocampal astrocytes and clarify its mechanism.Methods·According to the time after propofol injection,twenty-four SD rats were randomly divided into three groups,i.e.0 min,45 min and 90 min group.Rats were administrated intraperitoneally with propofol (10 mg/mL,100 mg/kg body weight).The levels of glial fibrillary acidic protein (GFAP) and S100β mRNA in rat hippocampus were evaluated by realtime PCR.And cell viabilities and levels of GFAP mRNA were examined in primary cultured hippocampal astrocytes induced by 10 μmol/L propofol with or without 10 μmol/L extracellular signal-regulated kinase (ERK) inhibitor PD98059 pretreatment.Results·The mRNA levels of GFAP in the hippocampal tissue were (1.32±0.12) times (P=0.000) and (1.12±0.09) times (P=0.012) that in 0 min group,respectively,45 min and 90 min after injection of propofol.The mRNA levels of S100β in the hippocampal tissue were (1.14±0.11) times (P=0.005) and (1.05±0.10)times (P=0.284) that in 0 min group,respectively,45 min and 90 min after injection of propofol.The mRNA levels of GFAP and S100β were timedependently altered,first increasing,and then decreasing.In vitro,the cell viabilities (P=0.041) and levels of GFAP mRNA (P=0.026) in primary cultured hippocampal astrocytes were significantly elevated after propofol treatment,and these effects of propofol were reversed by ERK inhibitor PD98059.Conclusion·Propofol time-dependently upregulated the expression of GFAP and S100β via ERK signaling pathway in rat hippocampal astrocytes,so as to activate astrocytes.

Clinicopathological characteristics and prognosis of salivary gland secretory carcinoma / 临床与实验病理学杂志

Purpose To investigate the clinicopathological features and prognosis of salivary gland secretory carcinoma(SC). Methods A retrospective study was performed including reviewing the clinical documents and pathological sections of 10 cases of SC. Immunohistochemical EnVision study and histochemical staining were performed in the 10 cases. Fluorescence in situ hybridization (FISH) was used to detect the ETV6-NTRK3 fusion gene. Results There were 8 men and 2 women included in this study. The mean age was 45 years(ranged from 22 to 62 years).9 cases occurred in the parotid and 1 case in submandibular gland. Local painless masses were common first symptoms. Tumor size ranged from 1 cm to 3.5 cm in maximum diameter (average 1.8 cm) and the cut surface of most tumors was solid with dusty red or grayish yellow color, of which 1 case showed cystic degeneration. Histologically, the tumors usually pushed and were invasive to the adjacent tissues. Sometimes, the tumors showed expansive growth pattern. Tumor cells demonstrated microcystic, cystic papillary and alveolar patterns. Eosinophilic secretions could be observed in both microcysts and lumen of alveolus. Immunohistochemically, 10 cases revealed diffuse positivity of cytokeratin, and 9 cases were diffusely and strongly positive for S-100, Mammaglobin, vimentin and CK7, whereas all cases were negative for CD117, Dog-1, p53, p63, SMA, and GATA3. The tumor cells were positive for PAS staining and negative for mucicarmine staining. The detection ETV6-NTRK3 fusion gene was carried out in 4 cases by FISH analysis, among which 3 cases were positive. Follow-up data were available in the 10 patients (ranged from 2 months to35 years), among which 9 patients were alive, except for 1 patient died of tumor recurrence and metastasis 16 years after surgery. Conclusion SC is a newly recognized rare malignant tumor of salivary gland with a lowgrade malignancy, slow growth pattern and favorable prognosis. The histological structures of microcysts and eosinophilic secretions are (he crucial histological characteristics of SC. Diffusely strong positive expression of S-100 and mammaglobin is helpful for the diagnosis and differential diagnosis of the tumor. The diagnosis of SC could be confirmed when ETV6-NTRK3 fusion gene could be identified.

Propofol activates rat hippocampal astrocytes time-dependently via ERK signaling pathway / 上海交通大学学报（医学版）

Objective: To detect the effects of propofol on rat hippocampal astrocytes and clarify its mechanism. Methods: According to the time after propofol injection, twenty-four SD rats were randomly divided into three groups, i. e. 0 min, 45 min and 90 min group. Rats were administrated intraperitoneally with propofol (10 mg/mL, 100 mg/kg body weight). The levels of glial fibrillary acidic protein (GFAP) and S100β mRNA in rat hippocampus were evaluated by realtime PCR. And cell viabilities and levels of GFAP mRNA were examined in primary cultured hippocampal astrocytes induced by 10 μmol/L propofol with or without 10 μmol/L extracellular signal-regulated kinase (ERK) inhibitor PD98059 pretreatment. Results: The mRNA levels of GFAP in the hippocampal tissue were (1.32±0.12) times (P=0.000) and (1.12±0.09) times (P=0.012) that in 0 min group, respectively, 45 min and 90 min after injection of propofol. The mRNA levels of S100β in the hippocampal tissue were (1.14±0.11) times (P=0.005) and (1.05±0.10) times (P=0.284) that in 0 min group, respectively, 45 min and 90 min after injection of propofol. The mRNA levels of GFAP and S100β were timedependently altered, first increasing, and then decreasing. In vitro, the cell viabilities (P=0.041) and levels of GFAP mRNA (P=0.026) in primary cultured hippocampal astrocytes were significantly elevated after propofol treatment, and these effects of propofol were reversed by ERK inhibitor PD98059. Conclusion: Propofol time-dependently upregulated the expression of GFAP and S100β via ERK signaling pathway in rat hippocampal astrocytes, so as to activate astrocytes.

Effects of propofol sedation on BDNF-TrkB/p75 signal and cognitive function in rat hippocampus / 上海交通大学学报（医学版）

Objective • To detect the effects of propofol sedation on cognitive function in rats and its mechanism. Methods • Forty-eight SD rats were randomly divided into three groups, i.e. control group, 100 mg/kg group and 300 mg/kg group. Rats were administrated intraperitoneally with propofol (10 mg/mL, 100 mg/kg or 300 mg/kg). The mRNA levels of brain derived neurotropic factor (BDNF)-TrkB/p75 signal molecules in rat hippocampus were evaluated by realtime PCR 45 min after propofol treatment. Learning and memory ability was examined by inhibitory avoidance (IA) test after propofol treatment. Results • The mRNA levels of BDNF in the hippocampal tissue were (1.20±0.13) fold (P=0.002) and (88±12)% (P=0.044) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group after injection of propofol. The mRNA levels of TrkB were (1.01±0.11) fold (P=0.982) and (86±11)% (P=0.018) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group. The mRNA levels of p75 were (1.02±0.10) fold (P=0.778) and (1.59±0.18) fold (P=0.000) of that in control group, respectively, in 100 mg/kg group and 300 mg/kg group. There was no significant difference of the 24 h IA memory retention latency between 100 mg/kg group and control group. The 24 h IA memory retention latency in 300 mg/kg group was significantly decreased compared with control group (P=0.028) and 100 mg/kg group (P=0.020). Conclusion • Propofol dose-dependently regulates the expression of BDNF-TrkB/p75 signal molecules, and high dose propofol may reduce cognitive function via BDNF-TrkB/p75 signal.

Value of combined detection of Cyclin D1 and claudin7 in differential diagnosis of renal epithelial tumors / 临床与实验病理学杂志

Purpose To investigate the value of Cyclin D1 and claudin 7 expression in the differential diagnosis of renal epithelial tumors.Methods The expression of Cyclin D1 and claudin7 was detected by immunohistochemical staining with tissue microarray.Totally 309 cases of renal epithelial tumors and 20 cases of normal renal tissues were collected for immunohistochemical staining.Comparative analysis for the values of Cyclin D1 and claudin 7 was performed on the diagnosis and differential diagnosis of renal tumors.Results The positive rates of Cyclin D1 in renal oncocytoma (RO),chromophobe renal cell carcinoma (ChRCC),clear cell renal cell carcinoma (CCRCC),papillary renal cell carcinoma (PRCC),Xp11.2 translocation/TFE3 gene fusion associated renal cell carcinoma (Xp11.2/TFE3) and clear cell tubulopapillary renal cell carcinoma (CCTPRCC) were 86.2％ (50/58),8.2％ (4/49),70.0％ (98/140),8.8％ (3/34),42.9％ (6/14),and 71.4％ (10/14),respectively.The positive rates of RO were significantly different from those of ChRCC and PRCC (x2 =64.72,52.56,P ＜0.000 1).Significant differences in positive rates of CCRCC and ChRCC (x2 =55.87,P ＜ 0.000 1),PRCC and Xp11.2/TFE3 (X2 =4.28,P=0.039),CCTPRCC and ChRCC (x2 =21.69,P ＜0.000 1)were also observed.The positive rates of claudin 7 in RO,ChRCC,CCRCC,PRCC,Xp11.2/TFE3 and CCTPRCC were 20.7％ (12/58),87.8％ (43/49),8.6％ (12/140),50％(17/34),14.3％ (2/14),and 57.1％ (8/14),respectively.There were significant differences in positive ratios of RO and ChRCC (x2 =47.82,P ＜ 0.000 1),CCRCC and CCTPRCC(x2 =26.57,P ＜0.000 1),PRCC and Xp11.2/TFE3 (x2 =5.29,P ＜ 0.05).The sensitivity and specificity of Cyclin D1 +/claudin 7-immunophenotype for RO were 69％ and 95.9％ respectively.The diagnostic sensitivity and specificity of Cyclin D1-/claudin 7 + for ChRCC were 83.7％ and 96.6％.In the identification of CCRCC and CCTPRCC,the sensitivity and specificity of Cyclin D1 +/claudin 7-for CCRCC were 65.7％ and 71.4％,and the diagnostic sensitivity and specificity of Cyelin D1 +/claudin7 + for CCTPRCC were 42.9％ and 95％.Conclusion The differential expression of Cyclin D1 and claudin7 in RO and ChRCC,CCRCC and ChRCC,PRCC and Xp11.2/TFE3 suggests that combined detection of two proteins provides an important assistance for the identification of these renal epithelial tumors.

Therapeutic Efficacy of VICP+L-ASP/TKI on Adult Patients with B-ALL / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic efficacy of VICP+L-ASP/TKI on adult patients with B-ALL and to explore the influence factors.</p><p><b>METHODS</b>Forty-one adult B-ALL patients treated with VICP+L-ASP/TKI from August 2008 to June 2014 were following-up. The complete remission(CR) rate, toxicity, overall survival(OS) and event free survival(EFS) after induction treatment were analyzed, the therapeutic outcome of patients between different risk stratification subgroups was compared, the influence of standardized consolidatory and maintaining treatment as well as allogeneic hematopoietic stem cell transplantation(allo-HSCT) on survival time was analyzed.</p><p><b>RESULTS</b>The early death not occurred in 41 patients with B-ALL including 37 cases with CR; the CR rate of 1 course treatment was 90.2%. The follow-up time lasted to March 17, 2015, the median follow-up time was 25(9-79) months; the 1 year OS rate was 75.3%, the EFS rate was 58.3%. Analysis of risk factors showed that the initial WBC count over 30×10/L, LDH over 250 U/L and minimal residual disease(MRD) over 10after treatment were poor prognostic factors. After remission, the standardized consolidatory treatment or allo-HSCT according to the "2012 China adult ALL diagnosis and treatment expert consensus" could improve long-term survival, 3 years OS rate was 73.8% and 61.5% respectively, 3 years EFS were 63.5% and 65.7% respectively. The main toxic and side effects were hematologic reactions, the hematologic adverse reaction of IV grade was observed in 97.6%(40/41) during induction treatment.</p><p><b>CONCLUSION</b>Induction chemotherapy based on VICP+L-ASP/TKI and standardized consolidatory after remission according to the "2012 China adult acute lymphoblastic leukemia diagnosis and treatment expert consensus" can improve the therapeutic efficacy. The allo-HSCT should be actively performed for B-ALL paients with high risk(elevated initial WBC count and LDH level); at some time, the regularly monitoring MRD and adjusting therapeutic protocol according to monitoring result can promote the prognosis of adult B-ALL patients.</p>

Plasmacytoid urothelial carcinoma of the urinary bladder: a clinicopathologic study of 16 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features and prognosis of plasmacytoid urothelial carcinoma (PUC) of the urinary bladder.</p><p><b>METHODS</b>The clinical and pathologic findings of 16 cases of PUC were retrospectively reviewed. Immunohistochemical study (MaxVision method) was carried out. The follow-up data were analyzed.</p><p><b>RESULTS</b>There were altogether 15 males and 1 female. The age of patients ranged from 40 years to 85 years (median = 64 years). Most patients (15/16) presented with hematuria. The tumor cells were small to medium in size and contained eccentric nuclei and moderate to abundant eosinophilic cytoplasm, assuming a plasmacytoid appearance. The architectural pattern varied from loosely cohesive sheets to cords, papillae, small nests or gland-like structures. Most tumors invaded into the lamina propria or muscularis propria. Twelve of the 16 cases had concurrent conventional urothelial carcinoma component. Immunohistochemical study showed that the tumor cells in all cases were strongly positive for AE1/AE3, epithelial membrane antigen, CK7 and CK18. CK20 and uroplakin III were also expressed in 9 cases. CEA, p53, CD138, p63 and E-cadherin were positive in 12, 13, 15, 11 and 10 cases, respectively. Ki-67 index ranged from 5% to 70% (mean = 30%). All tumors were negative for vimentin, LCA, kappa/lambda light chains, S-100 protein, HMB 45,Melan A, smooth muscle actin and desmin. Follow-up information was available in 13 patients. The duration of follow up ranged from 3 months to 10 years. Three patients died of distant metastasis at 3, 27 and 60 months after the operation, respectively. One patient was alive with disease at 25 months. One was alive at 43 months with a prior recurrence. Another 8 patients were alive and disease free at 7 to 120 months.</p><p><b>CONCLUSIONS</b>PUC of the urinary bladder is a rare variant of high-grade urothelial carcinoma. Immunohistochemical study with positivity for CK7, CK20, p63 and uroplakin III and negative staining for vimentin and LCA may be helpful in the differential diagnosis. PUC is a malignant tumor with high invasiveness, high recurrence rate and poor prognosis. Radical cystectomy is considered as the first line treatment for PUC.</p>

Expression of carbonic anhydrase IX, PAX2 and PAX8 and their association with clinicopathologic characteristics in renal epithelial tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the expression of carbonic anhydrase IX (CAIX), PAX2 and PAX8 in different types of renal epithelial tumor and their association with clinicopathologic characteristics.</p><p><b>METHODS</b>Immunohistochemical study by EnVision method was performed in order to assess the expression of CAIX, PAX2 and PAX8 in 155 cases of renal cell carcinoma and 4 cases of metastatic clear cell renal cell carcinoma (CCRCC). Ninety-six cases of non-neoplastic renal parenchymal tissue adjacent to CCRCC, 8 cases of clear cell hepatocellular carcinoma and 2 cases of clear cell hidradenoma were used as controls.</p><p><b>RESULTS</b>CAIX was commonly expressed in CCRCC (94.0%, 63/67), of which 77.8% (49/63) showed strong positivity. CAIX was focally positive in papillary renal cell carcinoma, collecting duct carcinoma and urothelial carcinoma of renal pelvis. It was negative in chromophobe renal cell carcinoma, oncocytoma and adjacent non-neoplastic renal tissue. CAIX was also strongly expressed in the 4 cases of metastatic CCRCC. Focal expression of CAIX was demonstrated in the 8 cases of clear cell hepatocellular carcinoma and 2 cases of clear cell hidradenoma. The expression of CAIX in CCRCC did not correlate with tumor grading, clinical staging and presence of distal metastasis. On the other hand, PAX2 showed positive expression in different types of renal epithelial tumor, clear cell hepatocellular carcinoma and clear cell hidradenoma in various degrees. In contrast, PAX8 was commonly expressed in all types of renal epithelial tumor, with the exception of urothelial carcinoma of renal pelvis. PAX8 was not expressed in clear cell hepatocellular carcinoma and clear cell hidradenoma. Regarding diagnosis of CCRCC, CAIX demonstrated high sensitivity and specificity. PAX2 showed high specificity but low sensitivity. PAX8 was sensitive and specific in the diagnosis of renal epithelial tumor.</p><p><b>CONCLUSIONS</b>CAIX is a useful immunohistochemical marker with high specificity and sensitivity in distinguishing CCRCC from other types of renal epithelial tumor and clear cell tumors of non-renal origin. PAX2 is a marker with high sensitivity and low specificity for diagnosis of renal epithelial tumors. PAX8 is typically expressed in renal epithelial tumors. The combined detection of CAIX, PAX2 and PAX8 is useful in the diagnosis and differential diagnosis of renal epithelial tumors.</p>

ITF-2357 on inhibition myeloid leukemic cell lines cells proliferation in vitro and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of ITF2357, a novel histone deacetylase (HDAC) inhibitor, on the growth, differentiation and apoptosis of acute myeloid leukemic (AML) cells and its mechanism.</p><p><b>METHODS</b>AML cell lines kasumi-1 cells as a model for AML1-ETO positive, and THP1 cells for AML1-ETO negative, the leukemic cells proliferation was analyzed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining and flow cytometry. AML1-ETO, acetyl-histone, and caspase protein was analyzed by Western blot.</p><p><b>RESULTS</b>0.5 µmol/L ITF2357 treatment significantly inhibited kasumi-1 cells proliferation, with the 48 h half inhibitory concentration (IC(50)) of 0.1 µmol/L. The initial inhibitory concentration of THP1 cell line was 5 µmol/L. ITF 2357 induced apoptosis of kasumi-1 cells in a time- and dose-dependent manner. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment by ITF2357. Early apoptosis cells increased from (1.44 ± 1.52)% to (24.51 ± 5.79)%. Late apoptosis cells increased from (2.37 ± 2.8)% to (63.66 ± 1.56)%. ITF2357 induced AML1-ETO degradation by caspase-dependent pathway. 0.25 µmol/L ITF2357 induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD13 and CD15. 5 µmol/L ITF2357 blocked the cells at G(0)/G(1) phase, G(0)/G(1) cells increased from (39.69 ± 6.56)% to (79.2 ± 6.51)% and s-phase cells declined from (60.12 ± 3.29)% to (18.97 ± 6.62)%. Kasumi-1 cells incubated with 0.5 µmol/L of ITF2357, AML1-ETO protein began to decrease at 24 hours and could hardly be detected at 96 hours. ITF2357 induced AML1/ETO degradation through a caspase-dependent mechanism. At the same time, acetylated H3 and H4 increased.</p><p><b>CONCLUSION</b>Low-dose HDAC inhibitor ITF2357 can effectively inhibit the AML cells proliferation, especially for AML1-ETO positive AML cells. It inhibits Kasumi-1 cells proliferation degradation of AML1-ETO protein expression, blocks the cells at G(0)/G(1) phase, and induces apoptosis and differentiation of the cells.</p>

Primary cardiac neoplasms: a clinicopathologic analysis of 81 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the disease spectrum, clinical and pathologic features of primary cardiac neoplasms at a single medical institution during a period of eight years.</p><p><b>METHODS</b>The clinical and pathologic features of 81 cases of primary cardiac neoplasms encountered at the Affiliated Hospital of Medical College, Qingdao University from January, 2004 to December, 2011, either for cardiovascular surgery or as pathology consultation cases, were retrospectively reviewed. Immunohistochemical study was carried out in selected examples.</p><p><b>RESULTS</b>Amongst the 81 cases studied, 73 cases (90.1%) were benign and 8 cases (9.9%) were malignant. Cardiac myxomas accounted for 97.3% (71/73) of all the benign cases. Six of the 8 malignant cases represented soft tissue sarcomas. The commonest presenting symptom was dyspnea. The histologic subtypes included myxoma (number = 71), angiosarcoma (number = 2), malignant fibrous histiocytoma (number = 2), hemangioma (number = 1), liposarcoma (number = 1), lymphoma (number = 1), myxofibrosarcoma (number = 1) and papillary fibroelastoma (number = 1).</p><p><b>CONCLUSIONS</b>While the majority of primary cardiac neoplasms are myxomas, other tumor types do exist. Accurate histologic diagnosis and timely surgery are crucial in patient management.</p>

Chromophobe renal cell carcinoma: a clinicopathologic study and immunophenotypes of 42 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features, immunohistochemical profiles and prognosis of chromophobe renal cell carcinoma (ChRCC).</p><p><b>METHODS</b>Forty-two cases of ChRCC were retrieved from the archival files of the Affiliated Hospital of Qingdao University, 401 Hospital of PLA and Qingdao Municipal Hospital from 2003 to 2011. The clinical and pathologic features of the tumors were reviewed. Hale colloidal iron staining was performed and EnVision immunohistochemistry was used to detect the expression of a series of immunologic markers. Forty cases of clear cell renal cell carcinoma and 10 cases of renal oncocytoma were selected as controls.</p><p><b>RESULTS</b>The patients included 17 males and 25 females. The age of patients ranged from 39 years to 78 years (median age = 57 years). On gross examination, the tumors ranged from 2 cm to 19 cm in greatest dimension (mean size = 7.3 cm). Histologically, the tumors were mainly composed of solid sheets, acini or tubules of malignant cells. The tumor cells contained clear finely reticular ("chromophobe") and eosinophilic cytoplasm with perinuclear clearing. The nuclear outline was irregular and wrinkled. Nucleoli were inconspicuous and mitotic figures were barely seen. Hale colloidal iron stain was positive in all cases. Immunohistochemically, the tumor cells were variably positive for EMA (100%, 42/42), CK7 (95.2%, 40/42), Ksp-cad (92.9%, 39/42), CK18 (88.1%, 37/42), CD117 (61.9, 26/42), CD10 (31.0%, 13/42) and PAX2 (28.6%, 12/42). They were negative for vimentin, CA IX and TFE3. The follow-up period in 31 patients ranged from 2 to 77 months (average duration = 29 months). Three patients died of tumor metastasis 3, 8, 13 months respectively after the operation. Twenty-eight patients were still alive without evidence of tumor recurrence.</p><p><b>CONCLUSIONS</b>ChRCC predominantly occurs in middle-aged and elderly patients. It often carries a favorable prognosis. The presence of plant cell-like morphology, pale cells with uniform reticular microvesicular appearance and perinuclear clearing are characteristic histologic features. The diffuse positivity for Hale colloidal iron stain and EMA/CK7/Ksp-cadherin/CD117-positive immunoprofiles are also useful in differential diagnosis.</p>

Generation of trans-arachidonic acid under nitrative stress is associated with upregulation of thromponsdin-1 in diabetic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Trans-arachidonic acids (TAAs), newly discovered markers of nitrative stress and the major products of nitrogen dioxide (NO2(·))-mediated isomerization of arachidonic acid (AA), represent a new mechanism of NO2(·)-induced toxicity. It has been reported that TAAs were generated in oxygen-induced microvascular degeneration model and TAAs were also generated in a diabetic retinopathy (DR) model. In this study, we examined high glucose-induced nitrative stress damage and TAAs levels and explored the possible mechanisms for DR caused by reactive nitrogen species.</p><p><b>METHODS</b>Diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ) at 60 mg/kg. Bovine retinal capillary endothelial cells (BRECs) were selectively cultured and incubated with normal or high glucose. The serum TAAs and AA in diabetic rats were measured by the gas chromatography and mass spectrometry (GC/MS) method. The ratio of peak area of TAAs to AA with selected ion of 79 was estimated by a group t-test. Thrombospondin-1 (TSP-1) in the rat retinas and BRECs extracts were examined by Western blotting. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) protein was examined by Western blotting in BRECs incubated with high glucose.</p><p><b>RESULTS</b>The TAAs to AA ratio (TAAs/AA) was significantly increased in the serum at 8, 12 and 16 weeks after STZ injection (P < 0.05), with no noticeable change found at 2 or 4 weeks (P > 0.05). Expression of TSP-1 in the retina of diabetic rats was progressively elevated according to the duration of diabetes. TSP-1 expression was increased in BRECs incubated with high glucose at 48 hours. Moreover, high glucose also increased ERK1/2 expression, which peaked at 30 minutes and then decreased in the following 48 hours.</p><p><b>CONCLUSION</b>An elevation of TAAs/AA is associated with high glucose-induced nitrative stress, which probably involves upregulation of TSP-1 through activating ERK1/2.</p>

Screening of phosphoprotein associated with glycosphingolipid microdomains 1 (PAG1) by cDNA microarray and influence of overexpression of PAG1 on biologic behavior of human metastatic prostatic cancer cell line in vitro / 中华病理学杂志

<p><b>OBJECTIVE</b>To screen for novel gene(s) associated with tumor metastasis, and to investigate the effect of overexpression of phosphoprotein associated with glycosphingolipid microdomains 1 (PAG1) on the biological behaviors of human prostatic cancer cell line PC-3M-1E8 in vitro.</p><p><b>METHODS</b>Four cDNA microarrays were constructed using cDNA library of prostatic cancer cells PC-3M-1E8 (high metastatic potential), PC-3M-2B4 (low metastatic potential), lung cancer cells PG-BE1 (high metastatic potential)and PG-LH7 (low metastatic potential)to screen genes which were differentially expressed according to their different metastatic properties. From a battery of differentially expressed genes, PAG1, which was markedly downregulated in both high metastatic sublines of PC-3M and PG was chosen for further investigation. Real-time PCR and Western blot were used to confirm the gene expression of PAG1 at mRNA and protein levels. Full-length coding sequence of human PAG1 was subcloned into plasmid pcDNA3.0 and the recombinant plasmids were stably transfected into PC-3M-1E8. The cell proliferation ability, anchorage-independent growth, cell cycle distribution, apoptosis rates and invasive ability were detected by MTT, and in addition, soft agar colony formation, flow cytometry analysis and matrigel invasion assay using Boyden chamber were also carried out respectively. All experiments contained pcDNA3.0-PAG1-transfected clones, vector transfected clones and non-transfected parental cells.</p><p><b>RESULTS</b>A total of 327 differentially expressed genes were obtained between the high and low metastatic sublines of PC-3M cells, including 123 upregulated and 204 downregulated genes in PC-3M-1E8. A total of 281 genes, including 167 upregulated and 114 downregulated genes were obtained in PG-BE1 cells. Nine genes were simultaneously downregulated and 8 genes were upregulated in both high metastatic cell lines of PC-3M and PG. The expression of PAG1 at mRNA and protein level were decreased in the high metastatic subline PC-3M-1E8. Western blot revealed that PAG1 protein was downregulated in PC-3M-1E8 cell line which was in agreement with the gene expression at mRNA level. The proliferation ability and clonogenicity of PAG1 overexpression cells by stable transfection were markedly decreased in comparing with that of the control cells (P < 0.05). Colonies formed in stably PAG1-transfected cells, the vector-transfected clones and parental cells were 26.7 ± 5.2, 47.2 ± 3.2 and 52.3 ± 3.4 respectively (P < 0.05). Flow cytometry analysis showed that the stable PAG1-transfected cells at G₀-G₁ phase were significantly more than that of the control cells (P < 0.05). However, no difference of the apoptosis rate was found between PAG1-transfected cells and control cells (P > 0.05). The number of cells passing through the matrigel and multipore membrane was also decreased in the stable PAG1-transfected cells (35.1 ± 4.9) compared with those of the vector-transfected clones (127.6 ± 6.6) and parental cells (135.0 ± 5.0, P < 0.05).</p><p><b>CONCLUSIONS</b>Using cDNA microarray technique and differential gene expression analysis of sublines of the parental cancer cell lines enable of revealing the metastasis-related genes, among which PAG1 represents one of those under-expressed genes in the high metastatic subline PC-3M-1E8. Transfection expression of PAG1 suppresses cell proliferation, anchorage-independent growth and invasive ability of PC-3M-1E8 cells in vitro. Conclusively, PAG1 may play an important role in inhibiting the proliferation, invasion and metastasis of the cancer cells.</p>

Influence of phosphoprotein associated with glycosphingolipid microdomains 1 on biologic behavior of human prostatic cancer cell line in-vitro / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of influence of phosphoprotein associated with glycosphingolipid microdomains 1 (PAG1) on the biological behavior of human prostatic cancer cell line PC-3M-1E8.</p><p><b>METHODS</b>The expression of GST-Raf1-RBD recombinant protein, a specific binding domain of active Ras (GTP-Ras), was induced by IPTG in JM109 bacterium. SDS-PAGE and coomassie brilliant blue staining were performed using cleaved product of the bacterium to determine the expression of the fusion protein. GST-pull down essay was designed to detect the level of active Ras in PGA1 and vector transfected, respectively and in the native PC-3M-1E8 cells. Western blotting was used to detect the expression levels of downstream proteins of Ras signal pathway which may be related to the function of PAG1. Phalloidine labeled by tetramethylrhodamine-5-(and-6) isothiocyanate (TRITC) was used for the staining of intracellular F-actin, Laser passing confocal microscopy was adopted for observing change of the cell morphology and the arrangement of F-actin.</p><p><b>RESULTS</b>After IPTG induction, the GST-Raf1-RBD recombinant protein, with a molecular weight of 33 000, was noticed to be highly expressed in JM109 bacterium. GST-pull down assay revealed that the expression level of active Ras markedly decreased after PAG1-transfection while the total Ras remained unchanged. The expression of p-ERK and cyclin D1 in the PAG1-transfected cells decreased in accordance with the level of active Ras. However, the expression of p21(WAF1/CIP1) and p-Akt didn't show any variation. Additionally, the structure of F-actin was turbulent and the pseudopodia of cells diminished conspicuously after PAG1-transfection. There was a high expression of PAG1 in normal human prostate tissue, however, the positive rate of PAG1 immuno-staining decreased in cases of prostatic adenocarcinoma, correspondent with increasing of the grading index of cell differentiation established in the Gleason grading system for the diagnosis of prostate adenocarcinoma.</p><p><b>CONCLUSIONS</b>An over-expression of PAG1 in PC-3M-1E8 cells effectively suppresses the activation of Ras and ERK, as well as the cyclin D1 expression, leading to an inhibition of the proliferation ability of tumor cells. The turbulence of F-actin and reduction of pseudopodia of cells result in an impairment of cell mobility, invasiveness and metastatic capability. In human prostate and prostatic adenocarcinoma tissues, the expression of PAG1 is related to the cellular differentiation and malignancy.</p>

Analysis of chemotherapeutic results and prognostic factors of adult acute lymphoblastic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical characteristics of adult acute lymphoblastic leukemia (ALL), compare the efficacy of different induction regimens and analyze the prognostic factors.</p><p><b>METHODS</b>Data of 149 adult ALL patients hospitalized in our institute between June 1998 and December 2005 were retrospectively reviewed. The results were analyzed with the SPSS11.5 software.</p><p><b>RESULTS</b>1) Out of 133 patients available immunophenotype data, 118 (88.7%) were B-ALL and 15 (11.3%) T-ALL. Cytogenetic analysis was performed in 105 patients, 40 cases (38.1%) of them had a normal karyotype and 65 (61.9%) chromosome aberrations. 2) 149 patients completed the VDCP, VDLP or VDCLP induction therapies (at least 4 weeks treatment for each), 140 (93.7%) of them achieved complete remission (CR) with the first course CR rates of 80.8%, 92.3% and 81.4% , respectively (P=0.618). CR rates in patients after the induction regimens with or without asparaginase were 95.5% versus 92.1% (P=0.566). With a median follow-up of 14.5 (1-75) months, the median disease free survival (DFS) was 12 (1-74) months and median overall survival (OS) 17.5 (1-97) months. DFS of the three regimen groups at 3 and 5 years were 18.5% and 14.8%, 24.7% and 9.9%, 39.5% and 39.5%, respectively (P=0.0066). 3) COX regression analysis showed that the age (over 40 years), white blood cell (WBC) count ( > 40 x 10(9)/L) , t(9;22) (q34;q11)-positive and less than 4 courses consolidation chemotherapy were the unfavorable prognostic factors.</p><p><b>CONCLUSIONS</b>Most adult ALL patients are B-ALL and karyotype have more changed. More than 90% patients can achieve CR with induction regimens consisting of 4 or 5 drugs. Induction regimens containing L-asparaginase may not affect the CR rate, but can improve DFS and OS. Age and WBC at diagnosis, presence of t(9;22) (q34;q11) and the courses of post-remission treatment are important prognostic factors.</p>

Study on signal transduction pathway in differentiation and apoptosis of leukemia cells induced by heat shock protein inhibitor / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the signal transduction pathway in the differentiation and apoptosis of leukemia cells induced by heat shock protein 90 (HSP90) inhibitor 17-Allyl amide-17-demethoxygeldanamycin (17AAG).</p><p><b>METHODS</b>Kasumi-1 cells were treated with increasing concentrations or exposure time of 17AAG. The total kit protein (CD117), phosphorylated kit protein and its downstream signaling molecules were measured by Western blot analysis. Mutated kit protein from control and 17AAG-treated Kasumi-1 cells was immunoprecipitated and immunoblotted for associated chaperones.</p><p><b>RESULTS</b>Total kit protein and kit activity were decreased in 17AAG treated cells, but c-kit mRNA level was not. Total AKT protein and phospho-AKT, as well as phospho-STAT3 were rapidly down-regulated in Kasumi-1 cell after treatment with 17AAG. There was no change in total STAT3 protein. Immunoprecipitation showed that 1 microM 17AAG treatment for 1 hour caused kit associated HSP90 decrease and HSP70 increase.</p><p><b>CONCLUSION</b>17AAG-induced apoptosis of Kasumi-1 cells is associated with a decline in Asn822Lys mutated kit protein level and phosphorylated kit, and with a downregulation in its downstream activated signaling molecules involved in proliferation. AKT is a client protein of HSP90. The changes of kit associated HSP90 and HSP70 satisfy the circulation mode of molecular chaperone complex.</p>

Effectiveness analysis of HA based triple-drug regimen as induction chemotherapy in the treatment of acute myeloid leukemia and its relationship with karyotype / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the complete remission (CR) rate, disease free survival (DFS) and overall survival (OS) of de novo acute myeloid leukemia (AML) patients treated with HA based three drugs induction chemotherapy and to explore the impact of cytogenetic abnormalities on the prognosis.</p><p><b>METHODS</b>Two hundred and forty-three untreated de novo AML patients were treated with HA based three drugs induction therapy. CR rate, DFS and OS were calculated. One hundred and eighty-four patients who had karyotype results were divided into four or three groups according to SWOG or MRC criteria respectively. Differences in CR rate, DFS and OS among different groups were evaluated.</p><p><b>RESULTS</b>The CR rate of all the 243 cases was 77.4%. The median DFS of the 188 CR patients was 28.5 (ranged from 1.0 to 153.0) months, DFS rates at 3 and 5 years were 45.4% and 40.2% respectively. The median OS of the 243 patients was 18.4 (range from 0.5 to 154.0) months. OS rates at 3 and 5 years were 36.9% and 31.4% respectively. According to SWOG criteria, CR rate, median DFS and OS were 97.8%, 87.4 months and 89.0 months for the favorable group; 81.9%, 17.6 months and 22.3 months for the intermediate group; 61.5%, 9 months and 11.5 months for the adverse group; and 79.3%, 29.0 months, 19.9 months for the unknown group, respectively. The differences among the four groups were statistically significant (P < 0.001). According to MRC criteria, CR rate, median DFS and OS were 96.1%, 79.9 months, 72.2 months for the favorable group; 80%, 17.6 months, 19.7 months for the intermediate group; and 43.8%, 16.5 months, 12 months for the adverse group, respectively. The differences among the three groups were statistically significant excepting for DFS between intermediate and adverse groups.</p><p><b>CONCLUSIONS</b>HA based triple-drug induction regimens are highly effective in obtaining higher CR rate and longer survival time. Cytogenetics is the important prognostic factor for AML patients and SWOG karyotype subtyping criteria is more appropriate than that of MRC, the differences among the three groups being statistically significant.</p>

The heat shock protein 90 inhibitor induces apoptosis and differentiation of Kasumi-1 and its mechanisms / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of 17-allylamide-17-demethoxygeldanamycin (17AAG), a heat shock protein 90 (HSP90) inhibitor, on the growth, differentiation and apoptosis of leukemic Kasumi-1 cells.</p><p><b>METHODS</b>Kasumi-1 cells were treated with 17AAG at different concentrations in suspension culture. Cell proliferation was analysed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining, agarose gel electrophoresis and flow cytometry. KIT protein was analysed by Western blot and c-kit mRNA by RT-PCR.</p><p><b>RESULTS</b>17AAG treatment caused a dose-dependent inhibition of the cell proliferation with the IC(50) of 0.62 micromol/L. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment. 17AAG induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD11b and CD15, a progressive decline in S-phase cell fraction and an increase in G(0)/G(1) cells. When Kasumi-1 cells were incubated with 1 micromol/L of 17AAG, KIT protein began to decrease at 2 hours and KIT protein could hardly be detected at 20 hours, but c-kit mRNA was not decreased.</p><p><b>CONCLUSION</b>17AAG treatment of Kasumi-1 cells could lower KIT protein expression, inhibit cell proliferation, induce cell partial differentiation, apoptosis and accumulation in G(0)/G(1) phase.</p>