Immune suppressive signaling regulated by latent transforming growth factor beta binding protein 1 promotes metastasis in cervical cancer

Although metastasis is the major cause of death in cervical cancer, the mechanism of metastasis is still unclear. The mRNA expression and protein level of latent transforming growth factor beta binding protein 1 (LTBP1) were detected in tumor tissues and paracancerous tissues from in-house samples. Cell proliferation, cell cycle, migration, and in vivo metastasis were determined after LTBP1 was knocked down. Then, 13 drugs were screened, and the changes in cell apoptosis and proliferation and tumor metastasis were detected after drug treatment in shRNA cells. In our in-house samples, LTBP1 was lowly expressed in cervical cancer tissues. After LTBP1 knockdown, cell proliferation was increased, and the ability of in vitro migration and in vivo metastasis was enhanced. At the same time, the proportion of myeloid derived suppressor cells (MDSC) in situ increased, the proportion of T cells decreased, and transforming growth factor beta-1 (TGFβ1) signaling was activated. After carboplatin treatment, LTBP1 shRNA cell line apoptosis increased, metastasis in vivo was limited, and the proportion of MDSC in situ decreased. LTBP1 was lowly expressed in cervical cancer, and the inhibition of LTBP1 can improve the malignant degree of the tumor, and this process can be blocked by carboplatin.

Meta-analysis of external fixator and open reduction and internal fixation for the treatment of distal radius fracture / 中国骨伤

OBJECTIVE@#To compare clinical efficacy and safety of plate internal fixation (ORIF) and external fixator (EF) in treating distal radius fractures by Meta-analysis.@*METHODS@#From establishment of database to August, 2019, randomized controlled trial (RCT) about open reduction and internal fixation (ORIF) and external fixation (EF) in treating distal radius fractures was conducted by using computer-based databases, including CNKI, VIP, Wanfang Data, Medline, Cochrane library databases. Data extraction and quality evaluation of included study according to inclusion and exclusion criteria, RevMan 5.3 software was used to perform Meta-analysis. Palm angle, ulnar deflection angle, radius height, grip strength, ulnar variation, disabilities of arm, shoulder and hand (DASH) score, total complication rate, infection rate and tendon rupture between two groups were compared.@*RESULTS@#Totally 19 RCT were included with 1 730 patients, 873 patients in ORIF group and 857 patients in EF group. Meta analysis result showed that after operation at 12 months, there were no significant difference in radial height [@*CONCLUSION@#Compared with EF in treating distal radius fracture, ORIF has better clinical effects in postoperative complications, palm angle, ulnar deviation angle, ulnar variation rate and infection rate. While there were no significant difference between in DASH score, radial height, tendon rupture and carpal tunnel syndrome better EF and ORIF. For the patient pursue rapid recovery of function, ORIF is better choice.

Clinical features of acute kidney injury in patients with nephrotic syndrome and minimal change disease: a retrospective, cross-sectional study / 中华医学杂志(英文版)

BACKGROUND@#Minimal change nephropathy (MCD) is a common pathological type of nephrotic syndrome and is often associated with acute kidney injury (AKI). This study aimed to investigate the clinical characteristics and related factors of AKI in patients with MCD and nephrotic syndrome.@*METHODS@#Patients from Chinese People's Liberation Army General Hospital who were diagnosed with pathological renal MCD with clinical manifestations of nephrotic syndrome were included from January 1, 2013 to December 31, 2017. Patients diagnosed with membranous nephropathy (MN) by renal biopsy from January 1, 2013 to December 31, 2017 are included as a control population. We retrospectively analyzed the clinical and pathological characteristics of patients as well as the percentages and clinical characteristics of AKI in different age groups. We assessed the correlation of pathological characteristics with serum creatinine using multivariate linear regression analysis.@*RESULTS@#A total of 367 patients with MCD were included in the analysis, with a sex ratio of 1.46: 1 (male: female) and an age range of 6 to 77 years. Among all the patients, 109 developed AKI (29.7%), and of these patients, 85 were male (78.0%). In the 586 patients with MN, 27 (4.6%) patients developed AKI. The percentage of AKI in MCD patients was significantly higher than that in MN patients (χ2 = 41.063, P < 0.001). The percentage of AKI increased with age in the MCD patients. The percentage of AKI in patients aged 50 years or older was 52.9% (46/87), which was significantly higher than that [22.5% (63/280)] in patients under 50 years (χ2 = 6.347, P = 0.013). We observed statistically significant differences in age (43 [27, 59] years vs. 28 [20, 44] years, Z = 5.487, P < 0.001), male (78.0% vs. 51.4%, χ2 = 22.470, P < 0.001), serum albumin (19.9 ± 6.1 g/L vs. 21.5 ± 5.7 g/L, t = 2.376, P = 0.018), serum creatinine (129.5 [105.7, 171.1] μmol/L vs. 69.7 [57.7, 81.9] μmol/L, Z = 14.190, P < 0.001), serum urea (10.1 [6.2, 15.8] mmol/L vs. 4.7 [3.6, 6.4] mmol/L, Z = 10.545, P < 0.001), IgE (266.0 [86.7, 963.0] IU/ml vs. 142.0 [35.3, 516.5] IU/ml, Z = 2.742, P = 0.007), history of diabetes (6.4% vs. 1.2%, P = 0.009), and history of hypertension (23.9% vs. 5.1%, χ2 = 28.238, P < 0.001) between the AKI group and the non-AKI group. According to multivariate linear regression analysis, among the renal pathological features analyzed, renal tubular epithelial cell damage (β = 178.010, 95% CI: 147.888-208.132, P < 0.001) and renal interstitial edema (β = 28.833, 95% CI: 11.966-45.700, P = 0.001) correlated with serum creatinine values.@*CONCLUSIONS@#The percentage of AKI in MCD patients is significantly higher than that in MN patients. Patients over 50 years old are more likely to develop AKI. Renal tubular epithelial cell injury and renal interstitial edema may be the main pathological lesions that are associated with elevated serum creatinine in patients with MCD.

Observation of tigecycline treatment for children with severe infection / 中国实用儿科杂志

OBJECTIVE: To evaluate the efficacy and safety of tigecycline therapy in children with severe infection.METHODS: We conducted a restrospective chart review of 114 children with severe infection in a tertiary hospital from May 1,2012 to April 30,2017. Inclusion criteria: receiving tigecycline administration for at least 2 days(4 doses). Clinical data and laboratory results were recorded before and after the therapy. RESULTS: Totally 114 children were enrolled,including 47 patients diagnosed with Acinetobacter baumanmii infection,with 52 Acinetobacter baumanmii strains. The in-hospital mortality was 23.4%. Median duration of tigecycline treatment was 13 days(2.5-13.5 days). Median duration of antibiotics prior to tigecycline treatment was 9 days(2-27 days). The total clinical improvement rate was 47.3%,and the etiological eradication rate was 38.9%. After treatment 24 cases got clinically improved in 47 patients and 26 strains were eradicated. No serious adverse effect was reported. CONCLUSION: The efficacy and safety of tigecycline should not be overvalued. Additional data from randomized controlled trials are required to assess the administration of tigecycline.

Reviews on Chemical Compositions and Pharmacological Effect of Gelsemium elegans / 中国实验方剂学杂志

Gelsemium elegans,the evergreen woody vine of the genus Gelsemium of the Loganiaceae family in China,is a traditional Chinese medicinal plant. It is spicy,bitter,warm,highly toxic and commonly used for dispelling wind-evil,attacking poison,reducing swelling and relieving pain. In this article, the researches on the chemical constituents and pharmacological effects of G. elegans in recent years were retrieved,reviewed and summarized. So far,alkaloids,iridoids,triterpenes,phenolic acids,steroids,coumarins,lignans,megastigmane glycosides and other ingredients have been separated from G. elegans. Alkaloids, mostly of indole alkaloids,are the main active ingredients there of which can significantly inhibit central nervous activity. Modern pharmacological studies have shown that alkaloids have a variety of pharmacological activities,which can achieve the purpose of anti-tumor by regulating cell cycle; enhance macrophage phagocytosis,protect white blood cells,and promote immune regulation;relieve cancer pain and long-term pain;reduce the myocardial contractility,get vasodilation to achieve antihypertensive effect;and also play an important role in the treatment of anxiety disorder and dermatosis. In the future research on G. elegans,it is still necessary to further study the chemical constituents,develop promising lead compounds,conduct in-depth research on its toxicology and clinical pharmacology,and clarify its mechanism of action,make it used more fully and reasonably,and lay the foundation for the application and safe use of G. elegans.

Effect of Montelukast on T-lymphocyte subsets and advanced oxidation protein products in immune thrombopenic purpura model mice / 中国免疫学杂志

Objective:To investigate the effect of Montelukast on T-lymphocyte subsets, cytokines and advanced oxidation protein products ( AOPP ) in immune thrombopenic purpura ( ITP ) model mice. To analyze the principle of the treatment by Montelukast. Methods: Forty ITP mice were randomly divided into control group,model group,Montelukast low dose group(3 mg/kg) and Montelukast high dose group(12 mg/kg). ITP model mice were successive administration for 14 days after building models for 7 days. Platelet counts,the index of thymus and spleen were calculated. T-lymphocyte subsets were detected by flow cytometry. IL-6,TNF-α,AOPP were detected by enzyme linked immunosorbent assays. Results: Comparison with control group,the PLT,thymus index and spleen index,CD8+,IL-6,TNF-α,AOPP of model group mice were significantly increased (P＜0. 05) while CD3+,CD4+,CD4+/CD8+were significantly decreased (P＜0. 05). Comparison with model group,PLT,thymus index and spleen index,CD8+,IL-6,TNF-α,AOPP of low dose group and high douse group mice were significantly decreased (P＜0. 05) while CD3+,CD4+,CD4+/CD8+were significantly increased (P＜0. 05). Conclusion: Montelukast can cure ITP regulate immune disorders,eliminate accumulation of AOPP and reduce level of IL-6 and TNF-α.

Effect of DEK targeting silence on proliferation and cell cycle of human hepatoma HepG2 cells / 新乡医学院学报

Objective To study the influence of targeted silencing of DEK on the proliferation and cell cycle of human hepatoma cell lines.Methods The human hepatoma cells line HepG2 were routinely cuhured and the cells were divided into blank control group,siRNA control group and DEK siRNA group when the cells grew to 90％ tusion.The cells in blank control group were cultured normally without any treatment;the cells in siRNA control group and DEK siRNA group were transfected with siRNA expression vector and DEK siRNA expression vector mediated by LipofectamineTM2000 liposomes,respectively.The expression of DEK mRNA in HepG2 cells was detected by real-time polymerase chain reaction;the expression of DEK and CyclinD1 protein in HepG2 cells was detected by Western blot;the proliferation of HepG2 cells was detected by methyl thiazolyl tetrazolium method,and the cell cycle was observed by flow cytometry.Results The expression of DEK mRNA in the blank control group,siRNA control group and DEK siRNA group was 0.826 ±0.052,0.776 ±0.051 and 0.420 ±0.050 respectively;the expression of DEK protein in the blank control group,siRNA control group and DEK siRNA group was 0.691 ± 0.073,0.726±0.061 and 0.311 ±0.038 respectively;the expression of CyclinDl protein in the blank control group,siRNA cuntrol group and DEK siRNA group was 0.712 ± 0.069,0.780 ± 0.074 and 0.434 ± 0.039 respectively.The expressions of DEK mRNA,DEK protein and CyclinD1 protein in DEK siRNA group were significantly lower than those in the blank control group and siRNA control group (P ＜ 0.05);there was no statistic difference in the expression of DEK mRNA,DEK protein and CyclinD1 protein between the blank control group and siRNA control group(P ＜0.05).The proliferation ability of HepG2 cells in DEK siRNA group after transfection of 24,48,72,96,120 h was significantly lower than that in the blank control group and siRNA control group(P ＜0.05);there was no statistic difference in the proliferation ability of HepG2 cells between the blank control group and siRNA control group at each time point(P ＜ 0.05).The proportion of G0 + G1 phase cells in DEK siRNA group was significantly higher than that in the blank control group and siRNA control group(P ＜ 0.05);the proportions of S phase and G2 + M phase cells in DEK siRNA group were significantly lower than those in the blank control group and siRNA control group(P ＜ 0.05);there was no statistic difference in the proportion of G0 + G1 phase,S phase and G2 + M phase cells between the blank control group and siRNA control group (P ＜ 0.05).The result of Pearson correlation analysis showed that the expression of CyclinD1 protein was positively correlated with the expression of DEK mRNA and protein(r =0.909,0.899;P ＜ 0.05).Conclusion DEK siRNA can inhibit the proliferation of HepG2 cells,and change the cell cycle distribution through down regulating the expression of DEK gene in HepG2 cells.This process may be related to the down regulation of the expression of CyclinD1.

First Report in China on the Identification and Drug Sensitivity of Mycobacterium elephantis Isolated from the Milk of a Cow with Mastitis / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In this study, milk from a cow with mastitis was analyzed to determine the presence of mycobacterial infection. Milk quality and security problems pertaining to the safe consumption of dairy products were also discussed in this study.</p><p><b>METHODS</b>Milk was preprocessed with 4% NaOH. Then, mycobacteria were isolated from the milk sample on L-J medium. The isolate was identified using multiple loci Polymerase Chain Reaction (PCR) and multi-locus sequence analysis with 16S rRNA, sodA, hsp65, and ITS genes. The drug sensitivity of the isolate to 27 antibiotics was tested through alamar blue assay.</p><p><b>RESULTS</b>Smooth, moist, pale yellow colonies appeared on the L-J medium within a week after inoculation. Based on the results of multiple loci PCR analysis, the isolate was preliminarily identified as non-tuberculous mycobacteria. The 16S rRNA, SodA, hsp65, and ITS gene sequences of the isolate exhibited 99%, 99%, 99%, and 100% similarities, respectively, with those of the published reference strains of Mycobacterium elephantis (M. elephantis). The drug sensitivity results showed that the strain is resistant to isoniazid, p-aminosalicylic acid, and trimesulf but is sensitive to ofloxacin, rifampicin, amikacin, capreomycin, moxifloxacin, kanamycin, levofloxacin, cycloserine, ethambutol, streptomycin, tobramycin, rifabutin, ciprofloxacin, linezolid, cefoxitin, clarithromycin, and minocycline.</p><p><b>CONCLUSION</b>To the best of our knowledge, this study is initially to report the isolation of M. elephantis from the milk of a cow with mastitis in China.</p>

Effect and mechanism of danshensu on hepatitis B virus reverse transcriptase and antigen expression / 中国中药杂志

MTT assay was used in this study to investigate the inhibitory effect of danshensu on the activity of 2.2.15 cells among human hepatoma cell line (HepG2); indirect fluorescence labeling method was used to measure the changes of reactive oxygen levels in the cells; ELISA method was used to determine hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels in cellular supernatants; HBV DNA level was measured with fluorogenic quantitative PCR method. The inhibitory effect of danshensu on HBV RT(hepatitis B virus reverse transcriptase) was studied by using enzyme inhibition dynamics, and the effect of danshensu on secondary structure of HBV reverse transcriptase was monitored by using circular dichroism. The results showed that danshensu had a good inhibitory effect on the growth of HepG2.2.15 cells, with a half inhibitory concentration (IC₅₀) of (15.35±2.43) μmol•L⁻¹; danshensu could significantly inhibit HBsAg and HBeAg expressions, and showed an inhibitory effect on HBV DNA replication. In addition, danshensu was an effective inhibitor for HBV reverse transcriptase [IC₅₀ (21.32±2.43) μmol•L⁻¹]. The fluorescence labeling results showed that the reactive oxygen levels in the cells were increased with the increase of danshensu concentration. Circular dichroism analysis showed that danshensu could induce partial change of conformation of HBV reverse transcriptase and gradually increased α-helical content. These results indicated that danshensu could make the structure of the enzyme become closer by binding to HBV reverse transcriptase, which was not conducive to the formation of the active center, so it could finally decrease the activity of HBV reverse transcriptase. Such decrease in enzyme activity would directly affect the HBV DNA replication, and combined with the decrease of the antigen levels, the effect of danshensu on HBV was increased.

Testicular malignant Leydig cell tumor: A case report / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological features of testicular malignant Leydig cell tumor (TMLCT) and improve the non-invasive diagnosis of the disease.</p><p><b>METHODS</b>We retrospectively analyzed the clinicopathological data on a case of TMLCT, detected the circulating tumor cells (CTC) in the peripheral venous blood, and reviewed the related literature.</p><p><b>RESULTS</b>The patient, a 47-year-old male, underwent radical orchidoepididymectomy under general anesthesia. Postoperative pathology confirmed the lesion to be TMLCT, which was mainly composed of Leydig cells and suspected with vessel carcinoma embolus. Immunohistochemistry showed the tumor cells to be positive for α-inhibin, Ki67, CD30, vimentin, EMA, and PLAP, but negative for CK, CK7, S100, CD10, SMA, Des, AFP, hCG, CEA, CK19, CD117, Oct-4, LCA, CD20, Pax-5, CD3, and CD43. Two CTCs were detected in the peripheral venous blood. The patient received 3 courses of chemotherapy for retroperitoneal multiple lymph nodes metastasis post-operatively. Subsequent CT imaging manifested no obvious reduction of the retroperitoneal lymph nodes and consequently the patient again underwent retroperitoneal lymphadenectomy and cryoablation. At 8 months after treatment, CT examination revealed notably enlarged retroperitoneal lymph nodes with the right adrenal gland evidently invaded.</p><p><b>CONCLUSION</b>TMLCT is an extremely rare sex-gonad stromal tumor with high malignancy and poor prognosis, and CTCs may be used for its early diagnosis and prognostic prediction.</p>

One Case of Multiple Myeloma with Central Never System Infiltration / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyses and summarize a case of multiple myeloma with disseminated infiltration in central nervous system.</p><p><b>METHODS</b>The results of laboratorial examination and clinical data were analyzed and compared in the light of published literatures.</p><p><b>RESULTS</b>The headache and diplopia were caused by infiltration of multiple myeloma cells to the central nervous system. Unlike those reported in the literatures, this case was a rare case of disseminated infiltration inside the brain, and plasma cells were CD56+, this patient has not yet accepted any multiple myeloma-associated treatment as like that reported in the literatures. And different from cases reported, this patient showed a good response to the intrathecal chemotherapy.</p><p><b>CONCLUSION</b>Whether this good response is due to a heterogeneity of MM or effect of treatment-associated drug is still to be decided.</p>

Expression and clinical significance of MMP-2 and MMP-9 in B acute lymphoblastic leukemia / 中国实验血液学杂志

This study was purposed to investigate the expression and clinical significance of MMP-2 and MMP-9 in patients with B-acute lymphoblastic leukemia (B-ALL). The expression of MMP-2 and MMP-9 in bone marrow mononuclear cells of B-ALL patients and normal controls was detected by RT-PCR. The gelatinolytic activity was detected by zymography. The results showed that the expression of MMP-2 in de novo and relapsed B-ALL patients was markedly higher than that in normal controls (P < 0.05). The expression of MMP-9 in de novo and relapsed B-ALL patients was markedly lower than that in normal controls (P < 0.05). The expression of MMP-2 and MMP-9 in patients with extramedullary infiltration was significantly higher than that in patients without extramedullary infiltration. The incidence of extramedullary infiltration in patients with MMP-2/MMP-9 (+) was markedly higher than that in patients with MMP-2/MMP-9 (-). The expression of MMP-9 was markedly higher in high-risk patients than that in standard-risk patients (P < 0.05), but the expression of MMP-2 had no significant difference between the high-risk and standard-risk patients (P > 0.05). It is concluded that MMP-2 and MMP-9 may be secreted by B lymphoblasts and may involve in the extramedullary infiltration. MMP-9 may correlate with poor prognosis.

Evolutionary analysis of neuraminidase gene of A/H7N9 influenza virus / 病毒学报

In 2013, the World Health Organization reported the first case of human infection with a new influenza A (H7N9) virus in China. This has caused damage and panic within certain areas in China. Therefore, analysis of this virus with bioinformatics technology is very necessary. Neuraminidase (NA) is one of the most important antigens of the influenza virus and an important target for anti-flu drugs. In this study, the nucleotide and protein sequences of NA gene of A/H7N9 influenza viruses were retrieved from the NCBI database, and MEGA 5.0 software was employed to construct a phylogenetic tree based on the nucleotide coding sequence; BioEdit software was used to align the nucleotide and protein sequences of NA and calculate the homologies of nucleotides and amino acids and then to analyze the important mutation sites of NA gene. The results demonstrated that the spread of influenza virus H7N9 showed certain geographical and temporal relations. The H7N9 virus isolated from China in 2013 belonged to Euroasiatic serotype, and its NA stalk region hadobvious variation, which may be one of the reasons that this virus infects human. These analyses may be very helpful for understanding the evolutionary relationship and mutation trend of A/H7N9 influenza viruses.

MNPs-Fe₃O₄mediates malignant Hematolpoectic cell apoptosis / 中国实验血液学杂志

This study was purposed to evaluate whether the safe concentration of magnetic nanoparticles of Fe₃O₄(MNPs-Fe₃O₄) for monocytes could induce the SKM-1 cell apoptosis. The average size and Zeta potential of MNPs-Fe₃O₄were determined by transmission electron microscopy and the Malvern Zetasizer 3000 HS, respectively. The cell viability after being exposed to MNPs-Fe₃O₄for 12, 24, 48, and 72 hours was detected by using cell count Kit-8. The cell apoptosis was evaluated by flow cytometry with Annexin V/PI double staining and Wright-Giemsa staining. The cell cycle was measured by flow cytometry. The levels of active caspase-3, survivin and bcl-rambo in cells treated with MNPs-Fe₃O₄and/or trolox for 48 hours were detected with Western blot. The results showed that the cell viability decreased in SKM-1 cells after exposure to 50 µmol/L and 100 µmol/L MNPs-Fe₃O₄(P < 0.05), but did not in monocytes (P > 0.05), compared with that of each non-MNPs-Fe₃O₄-treated group. This exposure also induced the SKM-1 cells to be arrested in G0/G1. Annexin V/PI staining assay showed that cell apoptotic rate induced by 100 µmol/L MNPs-Fe₃O₄was significantly high in SKM-1 cells while not so high in monocytes, and the pretreatment with trolox could attenuate the apoptosis. Moreover, the active caspase-3 increased in SKM-1 cells after the exposure to MNPs-Fe₃O₄, while that was not in monocytes, and the increased expression of BCL-rambo and the decreased expression of survivin involved in the process were also observed. It is concluded that MNPs-Fe₃O₄can induce the caspase 3-dependent SKM-1 cell apoptosis by increasing the BCL-rambo expression and decreasing the survivin expression, but this cytotoxic effect can not be observed in monocyte's.

Effects of Baicalin on an orthotopic transplantation mouse model of mismatch repair gene deficient colorectal cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To study the anticancer effects of Baicalin on an orthotopic transplantation mouse model of mismatch repair gene deficient colorectal cancer.</p><p><b>METHODS</b>Sixty orthotopic transplantation mice model of human colon cancer cell line HCT-116 expressing eGFP were established, which were divided randomly into negative controlled group (5% NaHCO3) and 50, 100, 200 mg/kg Baicalin groups. The nude mice were treated with intragastric infusion twice a day. Nude mice growth state, average weigh, inhibition rate of transplanted tumor, tumor metastasis and survival state were observed.</p><p><b>RESULTS</b>At 14, 21 and 28 days after treatment with different dose of Baicalin, tumor growth velocity was significantly slower in the treatment groups, and tumor volume was significantly smaller than the controlled group (there were (832 ± 637), (2012 ± 1566) and (2494 ± 1557) mm(3) respectively in 14, 21 and 28 days) (F = 4.433, P < 0.05). At the end point of study, survival state of 100 mg/kg group (13/15) was superior to controlled group (8/15) and 200 mg/kg group (8/15) (χ(2) = 4.665 and 3.980, P < 0.05).However, there were no significant differences in tumor metastasis and tumor surface vessel density.</p><p><b>CONCLUSIONS</b>Baicalin has statistically significant effects in inhibiting tumor growth in an orthotopic transplantation mouse model of mismatch repair gene deficient colorectal cancer, and 100 mg/kg may be an ideal treatment dose.</p>

Effect of SHIP mutation on invasion and migration of K562 leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of mutation in PxxP domain of SHIP on migration and invasion of leukemia cells and its mechanism.</p><p><b>METHODS</b>The lentiviral vector mediated wild type SHIP (wtSHIP) and mutant SHIP (muSHIP) plasmids were transfected into K562 cells through gene transfection techniques. Expression of SHIP at mRNA and protein level was detected by real-time PCR and Western blot, respectively. Transwell assay was used to analyze the difference between the migration and invasion ability of the K562/wtSHIP and the K562/muSHIP cells after transfection. Primary migration associated factor FAK, MMP and NF-κB were assayed by Western blot.</p><p><b>RESULTS</b>After transfection, the SHIP expression in transfected K562 cells were significantly increased. Compared with the migration ability of K562/wtSHIP\[(15.8 ± 1.4)%\], that of K562/muSHIP cells \[(54.3 ± 2.4)% \] increased greatly and almost at the same level of that of K562/pFIV\[(50.3 ± 3.8)%\] (P < 0.01). The invasion assay also showed that K562/wtSHIP\[(32 ± 6)/HP\] has a lower invasion ability than that of the K562/muSHIP group \[(83 ± 16)/HP\] and K562/pFIV group \[(78 ± 13)/HP\] (P < 0.01). Western blot analysis showed that the expression of p-FAK and NF-κB was up-regulated in K562/muSHIP group compared to that of the K562/wtSHIP group.</p><p><b>CONCLUSIONS</b>The results confirmed that mutation in PxxP domain of SHIP gene played an important role in negative regulating function of SHIP gene. The mutation affects the cell migration and invasion ability through increase in MMP-9 expression, FAK phosphorylation and NF-κB activation. It suggested that the mutation of PxxP domain in SHIP gene might be pathogenic, and be one of the reasons for SHIP abnormality in leukemia.</p>

Evaluation of 99Tcm-DTPA nuclear dynamic inaging in renal occupied disease / 肿瘤研究与临床

Objective To evaluate 99tcm-DTPA nuclear dynamic imaging in distinguishing the renal occupied disease.Methods A total of 164 in-patients with renal occupied disease who underwent surgery were included.According to the pathological diagnosis,119 patients had malignant tumors,and 45 patients had benign diseases.All patients’ imaging was retrospectively analyzed.Application of 99Tcm-DTPA nuclear dynamic imaging in renal occupied disease was compared with ultrasonography (US),computed tomography (CT),magnetic resonance imaging (MRI),intravenous pyelogram (IVP),and positron emission tomography (PET)-CT.Results The accuracy rates of different imaging methods in distinguishing between renal malignant and benign disease were 99Tcm-DTPA (84 ％,45 ％),US (72 ％,64 ％),CT ( 91％,92 ％),MRI (50 ％,67 ％),IVP (50 ％, 17 ％), respectively.The diagnostic accuracy rate of PET-CT for malignant tumors was 67 ％.The accuracy rates of 99Tcm-DTPA in distinguishing different phases of renal cell carcinoma were statistically significant (x 2 =83.4, P ＜ 0.01), while the accuracy rates in distinguishing renal cyst from renal angiomyolipoma were not statistically different.With the greater diameter, the diagnostic accordance rate is higher (x 2 =16.05,P ＜ 0.05).Conclusion 99Tcm-DTPA could be used not only to evaluate the renal function quantificationally,but also be helpful to distinguish renal malignant tumor from benign disease.

Effect of aging on pulmonary ICAM-1 and MCP-1 expressions in rats with lipopolysaccharide- induced acute lung injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of aging on the expressions of monocyte chemoattractant protein 1 (MCP-1) and intercellular adhesion molecule 1 (ICAM-1) in the lung tissue of rats with lipopolysaccharide (LPS)-induced acute lung injury (ALI).</p><p><b>METHODS</b>Both young (3 months old) and aged (27 months old) female Wistar rats were randomly divided into two groups (n=8), namely the normal control and LPS-induced ALI groups. Immunohistochemistry for of ED-1 was used to detect the infiltrating inflammatory cells. Western blot and Northern blot analyses were employed for evaluating the expressions of MCP-1 and ICAM-1 at the protein and mRNA levels.</p><p><b>RESULTS</b>Virtually no ED-1-positive cells were found in the lung tissue of the control rats in the young and aged groups. After LPS-induced ALI, ED-1-positive cells in the lung tissues increased significantly in both young and aged groups (P<0.05), and the increment was more obviously in the aged group (P<0.05). In the two normal control groups, the aged rats showed significantly higher expressions of MCP-1 and ICAM-1 than the young rats (P<0.05); LPS significantly up-regulated their expression in the young and aged groups (P<0.05), but the latter showed greater increments (P<0.05). The aged rats with ALI also showed significantly greater MCP-1 and ICAM-1 increments than those of the young rats (P<0.05).</p><p><b>CONCLUSIONS</b>Aging may upregulate lung MCP-1 and ICAM-1 expressions and enhance LPS-induced increments of MCP-1 and ICAM-1 expressions to exacerbate the pulmonary inflammation in rats.</p>

Effect of a methylation inhibitor 5-aza-2'-deoxycytidine on SHP-1 gene expression, proliferation and apoptosis in K562 cells / 中国实验血液学杂志

The aim of this study was to investigate the regulation of 5-aza-CdR on transcription of SHP-1 gene and effects on the proliferation and apoptosis of K562 cells. Methylation-specific PCR (MSP) was used to detect CpG island methylation in SHP-1 promoter. MTT and flow cytometry were used to detect the growth and apoptosis of K562 cells after treatment with different concentration of 5-aza-CdR. The expressions of SHP-1 mRNA and protein were determined by FQ-PCR and Western blot. The expression of p-JAK2 was assayed by Western blot. The result showed that methylation of SHP-1 gene promoter was detected in K562 cells, and the SHP-1 mRNA and protein were expressed again in K562 cells after treatment with 5-aza-CdR, meanwhile the expression of phosphorylated P-JAK2 was down-regulated; 5-aza-CdR significantly inhibited the cell growth in dose and time dependent manners. AG490 inhibited the cell proliferation. 5-aza-CdR increased the apoptosis rate of K562 cells also in dose- and time-dependent manners. The apoptosis rates of K562 cells treated with 5-aza-CdR for 1, 3 and 5 days were 9.3%, 24.2% and 37.7% respectively. After treatment with 2 micromol/L 5-aza-CdR for 24 hours, cells in G(0)/G(1) phase increased gradually, cells in G(2)/M phase decreased gradually, cells were arrested in G(0)/G(1) phase. The cell ratios in G(2)/M phase at 1, 3 and 5 days after treatment with 5-aza-CdR were 30.7%, 23.45 and 19.3% respectively. It is concluded that the 5-aza-CdR, inhibitor of specific methylation transferase, can re-express the silent SHP-1 gene in K562 cells, inhibits the proliferation of leukemia cells and induces cell apoptosis by activating JAK/STAT pathway.

The role of PTEN-FAK signaling pathway in metastasis and invasive ability of leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of the wild type phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a tumor-suppressor gene on the proliferation and apoptosis of human chronic myeloid leukemia (CML) cells line (K562) in vitro and explore the influence of PTEN-FAK signaling pathway on invasion and metastasis of leukemia cells.</p><p><b>METHODS</b>The recombinant Ad-PTEN gene containing green fluorescent protein gene (Ad-PTEN-GFP) or the empty vector (Ad-GFP) was transfected into K562 cells and fresh leukemia cells from CML patients in blast crisis. The growth of K562 cells was assayed by MTT assay; the apoptosis rate was assessed by flow cytometry (FCM). PTEN and FAK mRNA levels were detected by real-time fluorescent relative- quantification reverse transcriptional PCR (FQ-PCR) and its protein levels by Western blot. The metastasis and invasive ability was examined by transwell chamber assay.</p><p><b>RESULTS</b>The growth of K562 cells was suppressed markedly when Ad-PTEN-GFP was transfected into K562 cells at the 200 multiplicity of infection (MOI). The maximum growth inhibition rate was 35.2%. Transwell results showed the number of cells entered the lower chamber in Ad-GFP group was 9.1 fold more than that in Ad-PTEN-GFP group;The ability of metastasis and invasion of fresh leukemia cells was also suppressed after transfection with Ad-PTEN-GFP. FAK and p-FAK proteins were down-regulated by 0.72 and 0.16 fold lower after transfected with Ad-PTEN-GFP compared with Ad-GFP group.</p><p><b>CONCLUSIONS</b>PTEN gene might inhibit the proliferation, metastasis and invasive ability of leukemia cells via down-regulating FAK expression.</p>