Phylogenetic identification of symbiotic protists of five Chinese Reticulitermes species indicates a cospeciation of gut microfauna with host termites.

Symbiotic protists play important roles in the wood digestion of lower termites. Previous studies showed that termites generally possess host-specific flagellate communities. The genus Reticulitermes is particularly interesting because its unique assemblage of gut flagellates bears evidence for transfaunation. The gut fauna of Reticulitermes species in Japan, Europe, and North America had been investigated, but data on species in China are scarce. For the first time, we analyzed the phylogeny of protists in the hindgut of five Reticulitermes species in China. A total of 22 protist phylotypes were affiliated with the family Trichonymphidae, Teranymphidae, Trichomonadidae, and Holomastigotoididae (Phylum Parabasalia), and 45 protist phylotypes were affiliated with the family Pyrsonymphidae (Phylum Preaxostyla). The protist fauna of these five Reticulitermes species is similar to those of Reticulitermes species in other geographical regions. The topology of Trichonymphidae subtree was similar to that of Reticulitermes tree. All Preaxostyla clones were affiliated with the genera Pyrsonympha and Dinenympha (Order Oxymonadida) as in the other Reticulitermes species. The results of this study not only add to the existing information on the flagellates present in other Reticulitermes species but also offer the opportunity to test the hypotheses for the coevolution of symbiotic protists with their host termites.

The protective role and mechanism of liriodendrin in rats with myocardial infarction.

BACKGROUND: Liriodendrin is a therapeutic constituent of sargentgloryvine stem which is a famous Chinese traditional medicine. Previous studies have suggested liriodendrin could inhibit different pathways to treat inflammation in lung and intestinal tract. But whether it can treat myocardial infarction (MI) is unknown. We investigated the protective effect of liriodendrin on acute MI in rats and explored the specific mechanisms to expand the use of this traditional Chinese medicine. METHODS: The rats were randomized into the sham group (sham operation), control group (ligation of the left anterior descending artery), and liriodendrin group. The liriodendrin group was intragastrically administered with a liriodendrin solution (100 mg/kg). The control group and the sham group were intragastrically administered with normal saline. Before all rats were euthanized, echocardiography was used to detect their cardiac function. Hematoxylin and eosin (HE) staining and the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method were performed. Further quantitative detection of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) levels in tissues were also detected. Western Blot and real-time polymerase chain reaction (RT-PCR) were used to detect the apoptosis and nuclear factor kappa-B (NF-κB) pathway in tissues. H9C2 cells were used to detect the related mechanisms in vitro. RESULTS: Echocardiography showed that, compared to control group, the cardiac function of the liriodendrin group was significantly improved. histopathological staining of the control group showed that the myocardial tissue was severely damaged, and inflammatory cells were infiltrated. Compared to the control group, the apoptosis index of the liriodendrin group was significantly lower (P<0.05). Enzyme-linked immunosorbent assay (ELISA) results showed that the levels of IL-1ß and TNF-α in the control group were higher than those in the liriodendrin group (P<0.05). Meanwhile, apoptosis and the NF-κB pathway were inhibited after liriodendrin administration (P<0.05). Moreover, the mRNA transcriptional activity in the control group was also higher than that in the liriodendrin group (P<0.05). Because of the effect of liriodendrin, NF-κB pathway and apoptosis were downregulated in H9C2 cells which were exposed to ischemia-hypoxia. CONCLUSIONS: Liriodendrin may protect myocardial cells after myocardial infarction in rats by inhibiting the release of inflammatory factors, activation of the NF-κB pathway, and apoptosis.

[Improvement of cardiac function by recombinant adenovirus Ad-hBNP in rats with chronic heart failure].

OBJECTIVE: To evaluate the therapeutic effect of hBNP on rats with chronic heart failure (CHF). METHODS: Thirty CHF rats defined by echocardiography at 12 weeks post abdominal aortic constriction were randomly divided into Ad-hBNP group (2.5 × 10(10) VP/ml NS Ad-hBNP 1 ml/week × 4, n = 14), Ad-Track group (n = 8), placebo group (NS, n = 8), 10 sham-operated rats served as control group. After 4 weeks treatment, cardiac function was evaluated by echocardiography and hemodynamic measurements. Heart weight (HW) and HW/body weight (BW) ratio were determined. RESULTS: IVS, LVPW, LVEDD and LVESD were significantly reduced in the Ad-hBNP group [(2.34 ± 0.29) mm, (2.28 ± 0.18) mm, (6.50 ± 0.42) mm, (3.54 ± 0.59) mm] than those in the Ad-Track group [(2.71 ± 0.35) mm, (3.02 ± 0.85) mm, (7.71 ± 0.83) mm, (4.72 ± 0.80) mm] and in the NS group [(2.78 ± 0.23) mm, (2.83 ± 0.53) mm, (7.34 ± 0.97) mm, (4.55 ± 0.77) mm, all P < 0.05]. The LVEF and LVFS of the Ad-hBNP group [(79.27 ± 7.01)%, (43.38 ± 6.73)%] were significantly higher than in the Ad-Track group [(70.85 ± 4.81)%, (35.72 ± 3.68)%] and in the NS group [(69.67 ± 6.90)%, (34.91 ± 5.10)%, all P < 0.01]. HR [(417.48 ± 32.57) beats/min, (446.85 ± 61.49) beats/min, P < 0.05; (440.83 ± 32.18) beats/min, P < 0.05], LVEDP [(-4.24 ± 4.00) mm Hg (1 mm Hg = 0.133 kPa); (21.99 ± 6.80) mm Hg, P < 0.01; (18.00 ± 12.25) mm Hg, P < 0.01] were significantly decreased and while LVSP [(131.79 ± 15.76) mm Hg; (112.99 ± 32.35) mm Hg, P < 0.05; (117.13 ± 15.26) mm Hg], +dP/dt(max) [(5037.20 ± 430.41) mm Hg/s; (4217.40 ± 1354.15) mm Hg/s, P < 0.05; (4310.50 ± 1293.97) mm Hg/s, P < 0.05] and -dP/dt(max) [(-4382.00 ± 1304.79) mm Hg/s; (-3725.00 ± 791.34) mm Hg/s, P < 0.05; (-3890.00 ± 1043.73) mm Hg/s, P < 0.05]were significantly increased in Ad-hBNP group than in Ad-Track group and NS group (all P < 0.05). HW and HW/BW were also decreased in Ad-hBNP group than in the Ad-Track group and the NS group. CONCLUSION: Exogenous hBNP improved the cardiac function and attenuated remodeling in CHF rats.

Mechanism of action of resolvin D1 in inhibiting the progression of aortic dissection in mice.

BACKGROUND: To investigate the protective effect of resolvin D1 (RvD1) on aortic dissection (AD) in mice and explore the related mechanisms. METHODS: Mice were randomly divided into a blank group, model group, and RvD1 group. The RvD1 and model groups were administered 0.4% ß-aminopropionitrile (BAPN) solution, while the blank group was administered distilled water. When the experiment began, whether mice had AD was determined by echocardiogram. The RvD1 group was also administered RvD1 (30 µg/kg), while the model and blank groups were administered saline intraperitoneally. After 21 d, body weight trend and survival rate in the three groups were compared. The diameter of the ascending aorta of mice was detected by echocardiography. Then, the mice were sacrificed, and histopathological staining procedures were performed. Enzyme-linked immunosorbent assay (ELISA) was used to detect cytokines and chemokines in blood and tissue, respectively. RESULTS: At 21 d, there was no statistically significant difference in body weight between three groups (P>0.05). The survival rate showed a significant difference between the RvD1 and model group (P<0.05). Echocardiography revealed that compared with the RvD1 and blank groups, aortic dilatation was significant in the model group. Pathological staining showed that the destruction of the aortic wall structure and inflammatory cell infiltration were more noticeable in the model group than in the RvD1 group. A slight disintegration of elastic fibers and collagen in the aorta was observed in the RvD1 group, and the aortic structure was clear. The results of ELISA showed that the inflammatory factors levels in the RvD1 group, although higher than those in blank group, were significantly decreased compared with the model group. The ELISA results of AD tissue showed that at 21 d, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels in the aorta were significantly decreased in the RvD1 group compared with the model group (P<0.05). CONCLUSIONS: Administration of RvD1 significantly delayed aortic dilation and disintegration and inhibited local macrophage and neutrophil infiltration in the early stages of aortic injury. Moreover, RvD1 significantly downregulated the expression of cytokines and chemokines in aortic tissues and serum and improved aortic remodeling.

[Association of tryptophan hydroxylase gene A218C and serotonin transporter gene polymorphism with essential hypertension in Chinese northern Han population].

OBJECTIVE: To investigate the relationship between tryptophan hydroxylase (TPH) gene A218C in intron 7 and 5-hydroxytryptamine transporter (5-HTT) gene variable number tandem repeat (VNTR) in intron 2 and gene-linked polymorphic region (LPR) deletion/insertion polymorphism and essential hypertension (EH) in Chinese northern Han population. METHODS: A total of 280 EH patients and 200 normotensive controls were genotyped using polymerase chain reaction-restriction fragment length polymorphism technique. RESULTS: There were no significant differences in the frequencies of the genotypes and alleles of TPH gene A218C and 5-HTTVNTR between EH patents and controls (all P > 0.05). The genotype frequencies of SS, LS and LL in the 5-HTTLPR polymorphism was 68%, 29% and 3% in EH patients, 53%, 37% and 10% in the controls respectively (P < 0.01). The frequencies of allele S and L of the 5-HTTLPR were 82% and 18% in EH patients, 72% and 28% in the controls respectively (P < 0.01). Compared with the carriers of L allele (LS + LL), the EH risk was significantly higher in the SS homozygote (OR = 1.90, 95%CI = 1.31 - 2.77, P = 0.001). After adjustment of age, gender, body mass index, blood lipids, fasting blood glucose and blood uric acid level, the binary logistic regression analysis demonstrated that SS genotype in the 5-HTTLPR polymorphism was significantly related to occurrence of EH (OR = 1.47, 95%CI = 1.06 - 2.04, P = 0.021). CONCLUSIONS: The SS genotype of the 5-HTTLPR might be a susceptible gene to EH, while the TPH gene A218C and 5-HTTVNTR polymorphism is not associated with EH in Chinese northern Han population.

Differentially expressed long noncoding RNAs and regulatory mechanism of LINC02407 in human gastric adenocarcinoma.

BACKGROUND: Long noncoding RNAs (lncRNAs) have been identified to play important roles in the development and progression of various tumors, including gastric cancer (GC). However, the molecular role of lncRNAs in GC progression remains unclear. AIM: To investigate the differential expression of lncRNAs in human GC and elucidate the function and regulatory mechanism of LINC02407. METHODS: The Cancer Genome Atlas database was used to investigate the involvement of lncRNAs in GC. Quantitative real-time polymerase chain reaction was used to estimate the relative expression level of LINC02407 in GC tissues and cells. Functional experiments including CCK8 assay, apoptosis assay, wound healing assay, and transwell assay were used to investigate the effect of LINC02407 on GC cells. Some microRNAs were predicted and verified via bioinformatics analysis and the luciferase reporter system. Predictive analysis and Western blot assay were used to analyze the expression of related proteins. RESULTS: Many differentially expressed lncRNAs were identified in GC, and some of them including LINC02407 can affect the survival. LINC02407 was upregulated in tumor tissues compared with adjacent tissues. HGC-27 cells showed the highest LINC02407 expression and HaCaT cells exhibited the lowest expression. Different experiment groups were constructed using LINC02407 overexpressing plasmids and related siRNAs. The results of functional experiments showed that LINC02407 can promote the proliferation, migration, and invasion of GC cells but inhibit apoptosis. Luciferase reporter assay showed that hsa-miR-6845-5p and hsa-miR-4455 was downstream regulated by LINC02407. Western blot analysis showed that adhesion G protein-coupled receptor D1 (ADGRD1) was regulated by the LINC02407-miR-6845-5p/miR-4455-ADGRD1 pathways. CONCLUSION: LINC02407 plays a role in GC through the LINC02407-miR-6845-5p/miR-4455-ADGRD1 pathways, and thus, it may be an important oncogene and has potential value in GC diagnosis and treatment.

Protective effects of N(2)­L­alanyl­L­glutamine mediated by the JAK2/STAT3 signaling pathway on myocardial ischemia reperfusion.

To explore the protective effect of N(2)-L-alanyl-L-glutamine (NLAG) on myocardial ischemia-reperfusion injury (IRI), and observe the influence of NLAG on the Janus activated kinase signal transducer 2 and activator of transcription 3 (JAK2/STAT3) signaling pathway­associated molecules. Wistar rats were randomly divided into three groups: Sham, IRI and NLAG. In the IRI rat model, the cardiac hemodynamics, the maximum rate of left ventricular pressure (+dP/dtmax) and the left ventricular end­diastolic pressure (LVDP) were recorded. Hematoxylin­eosin and Masson staining were used to detect myocardial histological changes. The levels of plasma interleukin (IL)­1ß and ­6, tumor necrosis factor (TNF)­α, lactase dehydrogenase (LDH), troponin (cTn)I, creatine kinase (CK), heart type fatty acid binding protein (hFABP), malondialdehyde (MDA) and succinate dehydrogenase (SDH) were determined with ELISA. The protein expression levels of B­cell lymphoma (Bcl)­2, Bcl2­associated X protein (Bax), Caspase­3, JAK2, phosphorylated (p)­JAK2, STAT3 and p­STAT3 were detected by western blot analysis. The IRI model demonstrated notable myocardial injury; myocardial cells were arranged disorderly with some nuclei disappearing, and cardiac muscular fibers were degenerated. Following 60 min of reperfusion, LVDP, HR and +dP/dtmax were 31.3±4.53 mmHg, 239.17±8.45 beats/min and 615.17 mmHg/sec, respectively. Compared with the Sham group, the levels of LDH, cTnI, CK, hFABP release, inflammatory factors (IL­1ß, IL­6 and TNF­α) and oxygen free radical (MDA and SDH) levels were increased in the IRI group. In the NLAG group, myocardial injury was improved, the concentrations of LDH, cTnI, CK, hFABP, IL­1ß, IL­6, TNF­α, MDA were decreased, and SDH release was increased compared with the IRI group. In addition, NLAG significantly increased Bcl­2, JAK2, p­JAK2, STAT3 and p­STAT3 protein expression, and decreased Bax protein expression compared with the IRI group. In conclusion, myocardial ischemia­reperfusion can lead to myocardial cell apoptosis and myocardial injury and NLAG attenuates the IRI­induced mitochondrial oxidative stress injury and apoptosis by activating the JAK2/STAT3 signaling pathway, thus exerting protective effects against IRI.

Nanosizing of a drug/carrageenan complex to increase solubility and dissolution rate.

In this study, we present a novel approach of nanosizing a drug/polymeric complex to increase both solubility and dissolution rate of poorly water-soluble compounds. A hydrophilic polymer, lambda-carrageenan, was first complexed with a model poorly water-soluble compound to increase the compound's aqueous solubility. The compound/carrageenan complex was further nanosized by wet-milling to enhance the dissolution rate. By complexing with carrageenan, the compound became amorphous in the complex. Using additional carrageenan as a stabilizer for nanosizing, a nanosuspension of a compound/carrageenan complex with a median particle size of about 0.3 microm was successfully developed. The particle size of the nanosuspension did not increase significantly during the lyophilization process and was stable for at least 39 days at room temperature after lyophilization. This approach of nanosizing a drug/carrageenan complex increased the aqueous solubility of the compound from less than 1 microg/mL to 39 microg/mL. In addition to increasing aqueous solubility, a nanosized compound/carrageenan complex had a faster dissolution rate than the complex, the free compound, and the nanosuspension of the free compound.

Proprotein convertase subtilisin/kexin type 9 levels and aortic valve calcification: A prospective, cross sectional study.

OBJECTIVE: To investigate the possible association between plasma proprotein convertase subtilisin/kexin type 9 (PCSK9) and the incidence and severity of calcific aortic valve disease (CAVD). METHODS: This prospective, cross sectional study involved patients with and without (controls) aortic valve calcification diagnosed by transthoracic echocardiography and dual source computed tomography (DSCT) scan. Aortic valves calcification scores were calculated from DSCT scans and patients were graded: grade 1, no calcification; grade 2, mildly calcified; grade 3, moderately calcified; grade 4, heavily calcified. Plasma PCSK9 levels were measured using an enzyme-linked immunosorbent assay. RESULTS: Forty patients were grade 1 (controls), 32 were grade 2, 48 were grade 3 and 32 were grade 4. Plasma levels of PCSK9 were significantly different between the four groups and the highest value was observed in the patients with grade 2 calcification. Only low-density lipoprotein cholesterol and lipoprotein (Lp)(a) were associated with the severity of CAVD. Regression analysis showed that age, Lp(a) and PCSK9 were independent predictors of CAVD. CONCLUSION: Data from this cross sectional study in a small sample of patients showed that plasma PCSK9 was correlated with the presence of CAVD but not its severity.

[Therapeutic effects of combination of arsenic trioxide with low-dose all-trans retinoic acid on induction of remission acute promyeloeytic leukemia].

OBJECTIVE: To observe the therapeutic efficacy and side effects of arsenic trioxide (As2O3)combined with low-dose all-trans retinoic acid (ATRA) on remission induction in newly-diagnosed and relapsed patients with acute promyeloeytic leukemia (APL). METHODS: 224 patients of APL, 156 newly diagnosed patients, aged 34 (13 approximately 62), with a male/female ratio of 1.56, and 28 relapsed patients, aged, aged 34 (12 approximately 63), with a male/female ratio of 1.89, underwent As2O3 + ATRA therapy. The therapeutic effects was compared with that of As2O3 alone treatment on 40 newly diagnosed patients and 25 relapsed patients and that of ATRA alone treatment on 36 newly diagnosed patients and 15 relapsed patients. The treatment protocol for the combination group was as following: As2O3 was administered intravenously at a dose of 10 mg/day and ATRA was given orally three times per day at a dose of 10 mg. The complete remission (CR) rate, period to CR, incidence of early death and side effects were observed in the three groups. RESULTS: In the newly-diagnosed patients, there was no significant difference in CR rate among the three groups (92.5% for the As2O3/LD-ATRA group, 83.8% for the ATRA group, and 90% for the As2O3 group respectively). In comparison with As2O3 alone, administration of LD-ATRA to the patients in the As2O3/LD-ATRA group significantly shortened the period to CR (the medium time to CR was 28 days for the As2O3/LD-ATRA group and 39 days for the As2O3 group respectively). As compared to ATRA alone, treatment with As2O3 with low-dose ATRA showed a significantly lower incidence of early death (2.5% for the As2O3/LD-ATRA group and 13.9% for the ATRA group respectively). In the relapsed patients, the CR rate was significantly higher in the group treated with As2O3/LD-ATRA (71.4% for the As2O3/LD-ATRA group, 32.0% for the ATRA group, and 43.0% for the As2O3 group respectively). The combined use of LD-ATRA with As2O3 did not further enhance toxic side effects as compared to As2O3 alone or ATRA alone. CONCLUSION: As2O3/LD-ATRA regimen is superior to either regimen given alone to patients with APL. It is an efficient therapeutic approach to APL patients using a combination of As2O3 with low-dose ATRA.

Ultrasonographic features of triple-negative breast cancer: a comparison with other breast cancer subtypes.

BACKGROUND: Triple-negative breast cancer (TNBC) is known to be associated with aggressive biologic features and a poor clinical outcome. Therefore, early detection of TNBC without missed diagnosis is a requirement to improve prognosis. Preoperative ultrasound features of TNBC may potentially assist in early diagnosis as characteristics of disease. PURPOSE: To retrospectively evaluate the sonographic features of TNBC compared to ER (+) cancers which include HER(-) and HER2 (+), and HER2 (+) cancers which are ER (-). MATERIALS AND METHODS: From June 2012 through June 2014, sonographic features of 321 surgically confirmed ER (+) cancers (n=214), HER2 (+) cancers (n=66), and TNBC (n=41) were retrospectively reviewed by two ultrasound specialists in consensus. The preoperative ultrasound and clinicopathological features were compared between the three subtypes. In addition, all cases were analyzed using morphologic criteria of the ACR BI-RADS lexicon. RESULTS: Ultrasonographically, TNBC presented as microlobulated nodules without microcalcification (p=0.034). A lower incidence of ductal carcinoma in situ (p<0.001), invasive tumor size that is>2 cm (p=0.011) and BI-RADS category 4 (p<0.001) were significantly associated with TNBC. With regard to morphologic features of 41 TNBC cases, ultrasonographically were most likely to be masses with irregular (70.7%) microlobulated shape (48.8%), be circumscribed (17.1%) or have indistinct margins (17.1%) and parallel orientation (68.9%). Especially TNBC microlobulated mass margins were more more frequent than with ER (+) (2.0%) and HER2 (+) (4.8%) cancers. CONCLUSIONS: TNBC have specific characteristic in sonograms. Ultrasonography may be useful to avoid missed diagnosis and false-negative cases of TNBC.

MiR-300 regulate the malignancy of breast cancer by targeting p53.

OBJECTIVE: In this study, we investigated the role of miR-300 in regulating cell proliferation and invasion of breast cancer (BC) cells. METHODS: MicroRNA and protein expression patterns were compared between breast cancer tissue and normal tissue and between two different prognostic groups. The up-regulation of miR-300 was confirmed by real-time reverse transcription polymerase chain reaction and its expression was analyzed in MCF-7 breast cancer cells. RESULTS: We observed that miR-300 expression was frequently and dramatically up-regulated in human breast cancer tissues and cell lines compared with the matched adjacent normal tissues and cells. We further showed that transient and stable over-expression of miR-300 could promote cell proliferation and cell cycle progression. Moreover, p53, a key inhibitor of cell cycle, was verified as a direct target of miR-300, suggesting that miR-300 might promote breast cancer cell proliferation and invasion by regulating p53 expression. CONCLUSION: Our findings indicated that miR-300 up-regulation might exert some sort of antagonistic function by targeting p53 in breast cancer cell proliferation during breast tumorigenesis.

Association of annexin A2 with cancer development (Review).

Annexin A2 (ANXA2) is a well-known calcium-dependent phospholipid binding protein widely distributed in the nucleus, cytoplasm and extracellular surface of various eukaryotic cells. It has been recognized as a pleiotropic protein affecting a wide range of molecular and cellular processes. Dysregulation and abnormal expression of ANXA2 are linked to a large number of prevalent diseases, including autoimmune and neurodegenerative disease, antiphospholipid syndrome, inflammation, diabetes mellitus and a series of cancers. Accumulating data suggest that ANXA2 is aberrantly expressed in a wide spectrum of cancers, and exerts profound effects on tumor cell adhesion, proliferation, apoptosis, invasion and metastasis as well as tumor neovascularization via different modes of action. However, despite significant research, our knowledge of the mechanism by which ANXA2 participates in cancer development remains fragmented. The present review systematically summarizes the effects of ANXA2 on tumor progression, in an attempt to gain an improved understanding of the underlying mechanisms and to provide a potential effective target for cancer therapy.

Cutaneous metastasis of cholangiocarcinoma.

AIM: To investigate the clinical characteristics and prognostic factors of cutaneous metastasis of cholangiocarcinoma by a retrospective analysis of published cases. METHODS: An extensive search was conducted in the English literature within the PubMed database using the following keywords: cutaneous metastasis or skin metastasis and cholangiocarcinoma or bile duct. The data of 30 patients from 21 articles from 1978 to 2014 were analyzed. Patient data retrieved from the articles included the following: age, gender, time cutaneous metastasis occurred, number of cutaneous metastases throughout life, sites of initial cutaneous metastasis, anatomic site, pathology and differentiation of cholangiocarcinoma, and immunohistochemical results of the cutaneous metastasis. The assessment of overall survival after cutaneous metastasis (OSCM) was the primary endpoint. RESULTS: The median age at diagnosis of cutaneous metastasis of cholangiocarcinoma was 60.0 years (range: 35-77). This metastasis showed a predilection towards males, with a male to female ratio of 3.29. In 8 cases (27.6%), skin metastasis was the first sign of cholangiocarcinoma. Additionally, 18 cases (60.0%) manifested single cutaneous metastasis, while 12 cases (40.0%) demonstrated multiple skin metastases. In 50.0% of patients, the metastasis occurred in the drainage region, while 50.0% of patients had distant cutaneous metastases. The scalp was the most frequently involved region of distant skin metastasis, occurring in 36.7% of patients. The median OSCM of cholangiocarcinoma was 4.0 mo. Patient age and cutaneous metastatic sites showed no significant relation with OSCM, while male gender and single metastasis of the skin were associated with a poorer OSCM (hazard ratio: 0.168; P = 0.005, and hazard ratio: 0.296; P = 0.011, respectively). CONCLUSION: The prognosis of cutaneous metastasis of cholangiocarcinoma is dismal. Both male gender and single skin metastasis are associated with a poorer OSCM.

[Establishment of K562/NOD-SCID mouse model with leukemia].

The study was aimed to establish a K562/NOD-SCID leukemia mouse model and to explore its growth characteristics. Nude mice exposed to total body irradiation were inoculated subcutaneously with K562 cells, then the local K562 tumor was taken out and the tumor tissues without necoosis were selected for preparing single cell suspension which was inoculated into irradiated NOD/SCID mice by intraperitoneal injection. The results indicated that the systemic disseminated leukemia model was established successfully by intraperitoneal injection. On the fourth week after inoculation the leukemia cells were found on peripheral blood smear, and the leukemia cell infiltration was observed in liver, spleen and bone marrow. On the brink of death, the count of peripheral blood WBC was 8 - 10 times as much as that before inoculation. The leukemia cells on peripheral blood smear accounted to 20% - 30% of the total WBCs on average. The local tumors appeared in the abdominal cavity or on the greater omentum, less were involved in other organs. It is concluded that the established K562/NOD-SCID mouse model with leukemia well imitates the process of leukemia in human body, so it is a good model for the research on the effects of new drugs and target or gene therapy.

[Inhibition of leukemic cell proliferation by human soluble VEGF-R1].

The current study was purposed to investigate the inhibitory effect of human soluble vascular endothelial growth factor-1 (sFLT-1) on the proliferation of leukemic cells in vitro. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA and VEGF-R1 (FLT-1) mRNA in K562, HL60, U937 leukemic cell lines and bone marrow LTC-IC. Flow cytometry was used to detect the VEGF and VEGF-R1 (FLT-1) in all above-mentioned cells. VEGF concentrations in the cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Cell proliferation was determined by MTT after adding sFLT-1 to K562, HL60 and LTC-IC culture system. The result showed that expression of VEGF could be detected in K562, HL60, U937 leukemic cell lines and LTC-IC, especially K562, K562 and HL60 cell lines also expressed FLT-1, but a little expression was found in U937 and LTC-IC. sFLT-1 could effectively inhibit the growth of K562 and HL60 cell lines in dose-dependent manner. The highest inhibition rate was found at 48 hours after adding sFLT-1. It is concluded that sFLT-1 can inhibit the growth of some leukemic cell lines, and the inhibition effect enhances as the concentration of the sFLT-1 increase, but sFLT -1 not influence the proliferation of normal marrow cells.

[Monitoring of the therapeutic response of interferon to chronic myeloid leukemia by duel-color fluorescence in situ hybridization].

To investigate the significance of duel-color fluorescence in situ hybridization (D-FISH) in monitoring the response to interferon alpha (IFN-alpha) therapy in patients with chronic myeloid leukemia (CML), the D-FISH method was employed to detect the proportion of the interphase nuclei cells with bcr/abl fusion gene in the bone marrow of patients with CML before and after IFN-alpha therapy, and the results were compared with those of bcr/abl fusion mRNA by RT-PCR and Philadephia chromosome (Ph) by conventional cytogenetic analysis. The results showed that the mean detectable rate of bcr/abl fusion gene before and after IFN-alpha therapy was 96.4% and 58.6% respectively, in 22 patients who were bcr/abl-positive before IFN-alpha therapy by D-FISH method, was 94.0% and 70.1% respectively, in 2 patients of Ph-negative before treatment. Major, minor and no responses were seen respectively in 4, 4 and 14 cases from 22 patients by D-FISH method. The results also showed a good correlation with the analysis of RT-PCR and conventional cytogenetics. In conclusion, D-FISH method could directly detect the bcr/abl fusion gene of the interphase cells in bone marrow of patients with CML. It can overcome the defect of conventional cytogenetic methods which analyze only the cells in metaphase and the drawback of RT-PCR unable to quantify the bcr/abl fusion gene. D-FISH provides a more convenient and reliable method for evaluating the degree of clone remission to patients with CML after IFN-alpha therapy.

[Study on bone marrow microvascular density in hematological diseases].

To investigate the state and significance of bone marrow angiogenesis in hematological diseases, bone marrow microvascular density (BM-MVD) in plastic-embedded section was examined using acetone-fixed bone marrow tissues embedded in glycol-methacrylate (GMA) resin and by the method of immunohistochemistry. The results showed that bone marrow MVD increased greatly in newly diagnosed hematological malignancies before treatment. BM-MVD in patients with acute leukemia decreased down to the normal range as the controls at the time of complete remission. In the non-remission group, BM-MVD decreased less, but when relapsed it increased again up to the same range as the newly diagnosed hematological malignancies, significant increase of BM-MVD was found in patients with anemia, but in less degree than that in hematological malignancies. It is concluded that bone marrow angiogenesis plays a key role in the pathogenesis and development of hematological malignancy. Antiangiogenic therapy may be able to constitute a novel strategy for the treatment of hematological malignancies including leukemia.