Aim To study the effect of sibutramine hydrochloride on hypothalamus orexin system gene in high-fat dietary obesity rats.Methods SD rats were fed with high-fat diet to induce obesity model.Then,the rats were divided into two groups,the treatment group with sibutramine(8 mg?kg-1,ig)and control group with NaCl(ig)for 4 weeks.All rats' weight,blood fat and glucose were measured,prepro-orexin and its receptors mRNA expression in the hypothalamus were assessed by semi-quantity RT-PCR.Results Sibutramine not only decreased the obesity rats' weight and the level of serum lipid and glucose,but also increased the level of prepro-orexin mRNA expression in obesity rats.There was no difference in mRNA expression of both orexin receptors in each group.Conclusions Sibutramine hydrochlorid has an anti-obesity effect in high-fat obesity rats,which may be related to the increase of prepro-orexin gene mRNA expression level.

Fasting blood ghrelin level was decreased,and postprandial ghrelin descent was suppressed in high-fat diet induced obese rats.Fasting ghrelin concentration,but not postprandial ghrelin concentration,was negatively correlated with body weight,insulin resistance index,and glucose level.

Objective To observe glucose metabolism in C57 mice treated with different doses of fluoride.Methods Forty male C57 mice (body weight 20-24 g) were divided into four groups which were exposed to 0,50,100 and 150 mg/L sodium fluoride (NaF) by random number table according to body weight,each group had 10 mice.At 2,4,6,8,10 and 12 weeks after fluoride exposure,body weight was measured,blood glucose and glycosylated hemoglobin were detected by blood glucose meter and glycosylated hemoglobin meter,serum insulin and glucagon were detected by enzyme-linked immunosorbent assay (ELISA).Results At 10 and 12 weeks after fluoride exposure,the differences of fasting glucose between groups of C57 mice were statistically significant (F =35.12,21.92,all P ＜ 0.05),the fasting glucose of 100,150 mg/L fluoride groups [(7.7 ± 0.2),(7.3 ± 0.3),(8.6 ± 0.5),(9.1 ± 0.7)mmol/L] were higher than those of the control group [(5.4 ± 0.3),(5.0 ± 0.3)mmol/L,all P ＜ 0.01].The differences of glycosylated hemoglobin,glucagons between groups were statistically significant (F =3.85,8.74,all P ＜ 0.05).The glycosylated hemoglobin of 100,150 mg/L fluoride groups [(7.73 ± 0.76),(7.80 ± 1.15) mmo]/L] were higher than those of the control group [(5.43 ± 1.27) mmol/L,all P ＜ 0.05]; serum glucagon levels of 50,100,150 mg/L fluoride groups [(19.15 ± 11.84),(26.55 ± 15.97),(20.05 ± 7.29)ng/L] were lower than that of the control group [(48.35 ± 2.79)ng/L,all P ＜ 0.01].Conclusion Long term excess fluoride intake can reduce the function of sugar metabolism in C57 mice.

Objective To observe gene different expression of unfolded protein response signaling pathway in human osteoblasts under the excessive fluoride ,and explore the role of endoplasmic reticulum stress in fluorosis .Methods Human osteoblasts were cultured with fluoride ,intervening for 24 h .Cell viability and apoptosis were inspected by MTS assay and flow cytometer respective‐ly .The UPR signaling pathway was examined by real time PCR array ,and protein expressions were detected by Western blot .Re‐sults T he cell survival rates w ere (100 .678 5 ± 2 .830 3 )% ,(105 .393 4 ± 2 .538 4 )% ,(106 .125 7 ± 2 .048 3 )% ,(77 .977 3 ± 2 .544 3)% (P<0 .05) ,(30 .237 7 ± 0 .632 73)% (P<0 .05) treated with sodium fluoride at the concentration 0 ,5 ,10 ,20 ,40 ,80 mg/L respectively .Apoptosis rate inspected by flow cytometer was 4 .8% in 5 mg/L group ,13 .8% in 10 mg/L group ,37 .0% in 20 mg/L group ,58 .9% in 40 mg/L group ,63 .2% in 80 mg/L group (P<0 .05) .Only 1 gene was down regulated and 14 genes were up regulated .Western blot analysis showed BIP ,ATF4 ,CHOP and IRE1 both showed their protein expression gradually up regula‐ted with fluorine dose .XBP1 expression gradually increased in NaF 5-20 mg/L ,and its expression decreased at 40 and 80 mg/L . Conclusion Sodium fluoride can cause osteoblasts endoplasmic reticulum stress pathway through PTEN and IRE1 pathway ,and at high concentrations can cause apoptosis of osteoblast .

OBJECTIVETo optimize pre-coated multiple-probe fluorescence in situ hybridization (FISH) to improve its efficiency in cytogenetic diagnosis of acute leukemia.

METHODSThe original multiple-probe FISH techniques were optimized by adjusting the cell density and adding a process of protease digestion. Cytogenetic anomalies were detected in 141 patients with acute lymphocytic leukemia (ALL) or acute myeloid leukemia/ myelodysplastic syndromes (AML/MDS) using the modified technique, and 35 of the patients were also examined using the original technique. The successful detection rate and positive site detection rate were compared between the modified and original techniques.

RESULTSModification of the pre-coated multiple-probe FISH technique resulted in an significant increase of the successful detection rate (from 85.3% to 100%) and the positive site detection rate (from 5.1% to 8.6%) in ALL patients; in AML/MDS patients, the successful detection rate was significantly improved from 67.4% to 99.8% and the positive site detection rate from 3.5% to 6.0% (P<0.01).

CONCLUSIONThe modified pre-coated multiple-probe FISH technique can significantly increase the diagnostic efficiency of cytogenetic abnormalities in leukemic patients.

Chromosome Aberrations ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Myelodysplastic Syndromes ; diagnosis ; genetics

Objective: To analyze the association of cytogenetic abnormalities with the prognosis of chronic myeloid leukemia (CML) patients in tyrosine kinase inhibitors (TKI) era. Methods: Karyotype analysis of chromosome G-banding was carried out in 387 newly diagnosed CML patients by short-term culture of bone marrow cells. The correlation of cytogenetic abnormalities and CML progression was explored in combination with ABL tyrosine point mutations. Result: Of 387 patients with positive BCR-ABL fusion gene assayed by fluorescence in situ hybridization (FISH) technique, 94.1% (364/387) patients were Ph positive and 5.9% (23/387) Ph negative; 320 patients (87.9%) had a translocation t (9;22) (q34;q11) and 5 (1.4%) a variant translocation t (v;22) . Additional cytogenetic aberrations (ACA) at diagnosis were found in 10.7% (39/387) Ph⁺ patients, major route ACA in 22 (56.4%) cases and minor route ACA in 15 (38.5%) cases and 2 patients (5.1%) lacked the Y chromosome (−Y) ; 23.4% (71/303) patients occurred ACA during TKI treatment and the most frequent abnormalities were abnormal chromosome numbersd, which were likely associated with high proportion of disease progression (χ²=168.21, P<0.001) and ABL tyrosine point mutations (χ²=29.04, P<0.001) . Newly diagnosed CML-CP patients with t (9;22) (q34;q11) had a longer event-free survival (EFS) and disease-free survival (DFS) rates than that of patients with ACA (P=0.037; P=0.003) , while the overall survival (OS) had no significant differences (P=0.209) . As for CML-CP patients that occurred ACA during TKI therapy would have a marked low OS, EFS and DFS (all P<0.001) compared with no ACA occurred patients. Survival of advanced patients that occurred ACA were dramatically reduced. Conclusion ACA often emerged during the disease progress in CML patients, regular and timely detection of chromosomes karyotype and ABL tyrosine point mutations during TKI treatment was important for therapeutic evaluation, progress and prognosis of CML.

Objective: To investigate the clinical implications of p16 gene deletion in adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph⁺ ALL) . Methods: Retrospective analysis of clinical, immunophenotypic, cytogenetics, molecular characteristics and prognosis of 80 newly diagnosed Ph⁺ ALL patients with p16 deletion. Results: Of 80 adult Ph⁺ ALL, the prevalence of p16 gene deletion was 31.3%. p16 gene deletion carriers frequently accompanied with high WBC counts (WBC≥30×10⁹/L) and CD20 expression. The incidence of complex chromosome abnormality in p16 gene deletion group was higher than that in non-deletion group, with alternations in chromosome 7, 8, 19 and der (22) more frequently observed. There was no difference occurred between patients with or without p16 gene deletion in complete remission (CR) rate following induction chemotherapy combined with tyrosine kinase inhibitors (TKIs) . However, after three cycles of chemotherapy, the MMR and CMR rate in the p16 gene deletion group was lower than patients with wild-type p16 gene (P=0.034, P=0.036) . The p16 gene deletion patients showed no significant differences in MMR, CMR and relapse rate between Imatinib or Dasatinib plus chemotherapy (P>0.05) . Deletion of p16 gene was significantly associated with poor outcomes including worse overall survival (OS) (37.1% vs 54.1%, P=0.037) , lower disease free-survival (DFS) (12.4% vs 45.9%, P=0.026) , and increased cumulative incidence of relapse (P=0.033) . Among the 25 patients with p16 deletion, 14 underwent allo-HSCT and the median survival was 21 months, better than that of patients received chemotherapy alone (12 months) (P=0.030) . Conclusion This study indicated that deletion of p16 was associated with poor prognosis in adult Ph⁺ ALL, and the utility of second-generation TKI (Dasatinib) does not necessarily have an edge on efficacy over Imatinib, but allo-HSCT has the potential of elongating life expectancy. It is an important significance to define the status of p16 in Ph⁺ ALL for predicting prognosis and guiding therapy decision-making.

OBJECTIVETo investigate the cytogenetic differences between children and adults with acute lymphoblastic leukemia (ALL) using eight-probe fluorescence in situ hybridization and karyotype analysis.

METHODSEight-probe (MYC, P16, E2A, TEL/AML1, BCR/ABL , MLL , IGH, and hyperdiploidy) fluorescence in situ hybridization and karyotype analysis were performed for 86 adults and 39 children with acute lymphoblastic leukemia.

RESULTSEight-probe fluorescence in situ hybridization showed significant differences in the positivity rate of TEL/AML1, BCR/ABL, and hyperdiploidy between adult patients and children with ALL. By karyotype analysis, the positivity rate of t(9;22) and hyperdiploidy differed significantly between the children and adult patients (P<0.05).

CONCLUSIONAdults and children with ALL have different expression profiles of the fusion genes. Eight-probe fluorescence in situ hybridization is time-saving, accurate and efficient in detecting common genetic abnormalities in ALL patients, and can be well complementary to karyotype analysis in clinical diagnosis of ALL.

Adolescent ; Adult ; Child ; Child, Preschool ; Cytogenetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotype ; Karyotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; Young Adult

As essential part of teachers' teaching practice, the design and arrangement of assignments is the main carrier of interaction between teachers and students, and is also an important learning way for students. According to the discipline characteristics and teaching modes of histology and embryology courses, we have developed a systematic assignment design model that covers theory, practice, and application with four types of assignments—pre-class assignments, exploratory assignments, practical assignments, and innovative assignments distributed before class, in class, after class, and throughout the teaching process. Behind this assignment design strategy is the concept of learner-centered teaching, which aims to help students learn professional knowledge and develop comprehensive literacy.

OBJECTIVES: During pregnancy, pregnant women are prone to stress reactions due to external stimuli, affecting their own health and fetal development. At present, there is no good treatment for the stress reactions from pregnant women during pregnancy. This study aims to explore the effect of probiotics on abnormal behavior and hippocampal injury in pregnant stressed offspring. METHODS: SD pregnant rats were divided into a control group, a stress group, and a probiotics group, with 6 rats in each group. The control group was untreated; the stress group was given restraint stress on the 15th-20th day of pregnancy; the probiotics group was given both bifidobacterium trisporus capsules and restraint stress on the 15th-20th day of pregnancy, and the offspring continued to be fed with probiotics until 60 days after birth (P60). The offspring rats completed behavioral tests such as the open field test, the elevated plus maze test, the new object recognition test, and the barnes maze test at 60-70 d postnatally. Nissl's staining was used to reflect the injury of hippocampal neurons; immunohistochemical staining was used to detect the expression of microglia marker ionized calcium binding adapter molecule 1 (IBA-1) which can reflect microglia activation; ELISA was used to detect the content of plasma TNF-α and IL-1β; Western blotting was used to detect the expression of Bax, Bcl-2, and caspase-3. RESULTS: The retention time of offspring rats in the stress group in the central area of the open field was significantly less than that in the control group (P<0.01), and the retention time of offspring rats in the probiotic group in the central area of the open field was significantly more than that in the stress group (P<0.05). The offspring rats in the stress group stayed in the open arm for a shorter time than the control group (P<0.05) and entered the open arm less often than the control group (P<0.01); the offspring rats in the probiotic group stayed in the open arm for a longer time than the stress group and entered the open arm more often than the stress group (both P<0.05). The discrimination ratio for new to old objects in the offspring rats of the stress group was significantly lower than that of the control group (P<0.01), and the discrimination ratio for new to old objects in the offspring rats of the probiotic group was significantly higher than that of the stress group (P<0.05). The offspring rats in the stress group made significantly more mistakes than the control group (P<0.05), and the offspring rats in the probiotic group made significantly fewer mistakes than the stress group (P<0.05). Compared with the control group, the numbers of Nissl bodies in CA1, CA3, and DG area were significantly reduced in the offspring rats of the stress group (all P<0.001), the number of activated microglia in DG area of hippocampus was significantly increased (P<0.01), the contents of TNF-α and IL-1β in peripheral blood were significantly increased (P<0.05 or P<0.01), the protein expression level of Bcl-2 was significantly down-regulated, and the protein expression levels of Bax and caspase-3 were significantly up-regulated (all P<0.001). Compared with the stress group, the numbers of Nissl bodies in CA1, CA3, and DG area were significantly increased in the probiotic group offspring rats (P<0.001, P<0.01, P<0.05), the number of activated microglia in the DG area of hippocampus was significantly reduced (P<0.05), and the TNF-α and IL-1β levels in peripheral blood were significantly decreased (both P<0.05), the protein expression level of Bcl-2 was significantly up-regulated, and the protein expression levels of Bax and caspase-3 were significantly down-regulated (all P<0.001). CONCLUSIONS Probiotic intervention partially ameliorated anxiety and cognitive impairment in rats offspring of pregnancy stress, and the mechanism may be related to increasing the number of neurons, inhibiting the activation of hippocampal microglia, and reducing inflammation and apoptosis.
Animals ; Caspase 3/metabolism* ; Female ; Hippocampus/physiopathology* ; Humans ; Pregnancy ; Probiotics/therapeutic use* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Rats ; Stress, Psychological/therapy* ; Tumor Necrosis Factor-alpha/metabolism* ; bcl-2-Associated X Protein/metabolism*