BACKGROUND:Although tumor stem cel s and their differentiated vascular cel s, which are not sensitive to chemotherapy and molecular targeted therapy, reduce overal blood supply of the tumor, these cel s facilitate the invasion and metastasis of the tumor, eventual y resulting in treatment failure. OBJECTIVE:To investigate the effect of gastric cancer stem cel s in stomach cancer invasion and metastasis as wel as its mechanism in angiogenesis. METHODS:A total of 20 gastric cancer stem cel s and 20 normal gastric cancer cel s SGC7901 were cultured in the medium with DMEM/F12 (1:1), 2%B27 and 20μg/L epidermal growth factors or cultured in 10%PBS, respectively. Invasion of gastric cancer stem cel s was detected by sphere formation assay, colony formation assay and Transwel invasion assay. Levels of mRNA and protein expression of CD44 and E-cadherin were measured to observe tumor metastasis. Besides, angiogenesis was observed by cel scratch test, Transwel migration assay and ring test. RESULTS AND CONCLUSION:Gastric cancer cel s adhered to the medium wal , and gastric cancer stem cel s suspended in the medium, which appeared to be spindle interstitial form. Number of gastric cancer tumor spheres was significantly lower than that of gastric cancer stem cel spheres in the sphere formation assay and colony formation assay (P<0.05). Number of gastric cancer stem cel s through the basement membrane was significantly higher than that of gastric cancer cel s (P<0.05). And the cel migration distance and ring number were significantly increased in the gastric cancer stem cel s as compared with the gastric cancer cel s (P<0.05). In conclusion, the invasion, migration and angiogenesis abilities of gastric cancer stem cel s are stronger than those of gastric cancer cel s, which can further promote the occurrence and progression of gastric cancer.

Objective To investigate the effect of Mlot-4 cells onset with Roscovitine (ROSC)as some Cyclin-dependent kinases(CDK),inhibitor.Methods The logarithmic growth phase Molt-4 cells treated with ROSC at a final concentration ranging between 1～20 μmol/L and harvested in different time point,DNA assay of single cells by flow cytometry was used to detect the effect of cell cycle arrest and Annexin-V/FITC assay was used to detect the effect of cell apoptosis. Results It showed that ROSC exerted strong inhibitory effect on proliferation and cell cycle progression of Molt-4 Accumulation of G2/M arrested cells starting 6 h after onset of 10 μmol/L and 20 μmol/L ROSC;at the same time,the cell apoptosis of Molt-4 would be detected,According with the time and concentration changing,the cell apoptosis rate would rise.Conclusion It is concluded that Roscovitine(ROSC)as some Cyclin-dependent kinases(CDK),inhibitor,It has dual effects to Molt-4 cells,not only the effect of cell cycle arrest but also the effect of cell apoptosis.

Objective To explore the correlation between serum hs-CRP and β-cell finction in patients with gestational diabetes mellitus(GDM).Methods The levels of hs-CRP in 60 patients with GDM(GDM group)and 30pregnant women with normal glucose tolerance(NGT group)were detected.Insulin resistance was assessed by the homeostasismodel insulin resistance index(HOMA-IR),Insulin secretion by the homeostasis β-cell funetiOn index(HOMA-β).Results The levels of hs-CRP and HOMA-IR were higher in GDM group than in NGT group.There was significant difference between two groups(P＜0.01);HOMA-β was lower in GDM group than in NGT group,there was significant difference between two groups(P＜0.05).The level of hs-CRP was correlated with age,pre-pregnant bodymass-index(BMI),screening BMI,fasting blood glucose(FBG),fasting insulin(Fins),and HOMA-IR(r=0.222,0.649、0.862、0.923、0.935、0.941,P＜0.05 or P＜0.01),but was inversely related with HOMA-β(r=-0.872,P＜0.01).Multiple stepwise regression analysis indicated that HOMA-IR and HOMA-β was the most important effect factors of hs-CRP.Conclusion The level of hs-CRP increased in women with GDM.which was related to insulin resistance(IR)and insulin secretion,and it maybe participate in the pathogenesis of GDM.

Objective To investigate apoptosis of gastric cancer cells induced by taxol,and its specific apoptotic cell cycle phase. Methods The apoptosis rate of gastric cancer cell line MKN-28 induced by taxol was detected with Sub-G1 method.The specific apoptotic cell cycle phase Was detected with API and PSC method.The sensitivity of 20 cases clinic gastric cancer specimens induced by taxol was detected with the method MTT.Results With the Sub-G1 method the MKN-28 cells were induced by 10 μg/ml taxol,after 10 h,the apoptosis rate reached its top apex;with the API method and the PSC method,the specific apoptosts took place in G_2/M phase;the chemotherapy sensitivity of 16 cases out of 20 cases clinic gastnc cancer specimens exceed 50%with the method MTT. Conclusion Taxol could induce gastric cancer cells to aimptosis and the apoptosis takes place in G_2/M phase;Taxol is sensitive to clinic gastric cancer speclmens.

Objective To investigate the phosphorylation and dephosphorylation of CDK1 based on the specific cell cycle apoptosis in Molt-4 cells and active variety of CDK1 in cell cycle specific apoptosis.Methods The exponential phase of growth Molt-4 cells (the human acute lymphoblastic leukemia cell line) were induced with dose response and time course of Camptothecin (CPT).The specific cell cycle apoptosis was detected with API method,then cell apoptosis was verified with post sorting confocal method.Meanwhile,the phosphorylation and dephosphorylation of CDK1 were detected by the protein electrophoretic analysis (Western blot).Results The specific cell cycle apoptosis occurred on exponential phase of growth Molt-4 cells after CPT treatment.When Molt-4 cells occured S-phase apoptosis, the apoptosis cell phosphorylation of CDK1-Thr161 band was more narrow than that of control cells, the apoptosis cell phosphorylation of CDK1-Thr15 band almost had the same band with control cells.Conclusion Cell apoptosis frequently developed in different cell cycle phase. API assay is quick and efficient method to analyze specific cell cycle apoptosis. When cell apoptosis take place in S-phase,the phosphorylation activity of CDK1 is inhibited.

Objective To strengthen the voluntary blood donation publicity through the analysis of the voluntary blood donation.Methods The ratio of voluntary blood donation was statistically analyzed between staffs in some Medical institutions in Hangzhou and Ordinary people.Results The ratio of voluntary blood donation in ordinary people is 1.82％,which is higher than the national data (1.05％).The ratio of medical institutions in Hangzhou is 9.54％,which is far higher than ordinary people.The data of provincial,civic and private medical institutions are 10.20％,6.11％ and 15.42％,respectively.Conclusion The ratio of voluntary blood donation is different between different level hospitals,and the staffs in medical institutions are more willing to donate blood than ordinary people.

OBJECTIVE To study the drug-resistance of commonly encountered pathogens and the manifestations of irrational use of antibiotics,in order to provide the gist for clinical therapy.METHODS The retrospective analysis on the drug-resistance of commonly encountered pathogens that isolated from clinical samples and records of(irrational) use of antibiotics from Jan 2004 to Jon 2005 were carried out.RESULTS Among 870 isolates,the first five kinds of pathogens were Pseudomonas aeruginosa(17.47%),Klebsiella pneumoniae(9.66%),Acinetobacter baumannii(8.85%),Escherichia coli(7.36%)and Candida albicans(11.95%).They had resistance to the commonly used(antibiotics) in various degrees.In 4 462 records,there were 659 records of irrational use of (antibiotics),occupied 14.77%.CONCLUSIONS It is important that to strengthen the management of antibiotics(usage) and institutions in hospitals,to inform doctor the drug-resistance trend,based on the result of drug(sensitivity) test,in order to use antibiotics more rationally and reduce the appearance of drug-resistant bacteria.

AIM: To investigate the role of fluoxetine in the hippocampal synaptic plasticity in chronic unpre-dictable mild stress (CUMS) depression rats and its effect on mTOR and autophagy signaling pathways.METHODS:Male Sprague-Dawley rats (n =60) were randomly divided into normal control group, CUMS group and fluoxetine group. The CUMS rat model was established through CUMS combined with solitary raising, and fluoxetine (20 mg? kg -1? d -1 ) was administered via intragastric gavage.The changes of body weight, the ratio of sugar intake and the results of the behav-ioral test were recorded to identify the modeling.Moreover, the expression of synaptic plasticity-related proteins glial fibril-lary acidic protein (GFAP) and synaptophysin (SYP), apoptosis-related proteins Bcl-2 and caspase-3, mTOR signaling proteins mTOR and 4EBP1, and autophagy-related proteins beclin 1 and LC3 were examined by RT-PCR and Western blot. RESULTS: Compared with control group, the body weight, sucrose intake, and total distance and intermediate residence time in the open field test were significantly decreased in CUMS group.The results of RT-PCR and Western blotting showed that the mRNA and protein levels of SYP and GFAP in CUMS group were significantly down-regulated compared with con-trol group.The expression of Bcl-2 in CUMS group was downregulated, while the protein level of cleaved caspase-3 in-creased.Decreased phosphorylation levels of mTOR and its downstream target molecule 4EBP1 were observed in CUMS group.Besides, the autophagy-related proteins beclin 1 and LC3 were significantly upregulated at mRNA and protein lev-els.All these results(upregulation or downregulation) were attenuated by the treatment with fluoxetine, and the difference was statistically significant.CONCLUSION: Fluoxetine might improve hippocampal synaptic plasticity and alleviate symp-toms of depression by supressing apoptosis/autophagy signaling pathways and upregulating mTOR signaling pathway.

Objective To study the guiding role of ultrasonography to the operation of spontaneous constriction of radial nerve.Methods Six patients with spontaneous constriction of radial nerve were checked by ultrasound before operation to identify the number,position and morphological change of the constriction lesions of the radial nerve.The result was compared with the operation result.Results According to the ultrasonography results,2 to 3 constriction lesions were found on the radial nerve of all the 6 patients.The ultrasonography description of the number,position and morphological change of the lesions was consistent with the operation result.After the operation,the radial nerve function of all the patients got recovered.Conclusions Ultrasonography examination before operation can identify the number,position and morphological change of the constriction lesions of the radial nerve,which has great value when planning the operation.

Objective:To investigate the clinical features, the key point of diagnosis and treatment methods of X-linked hyper-IgM syndrome (XHIGM).Methods:The clinical characteristics and laboratory data of a patient aged 23 years who was diagnosed as XHIGM complicated with T-cell large granular lymphocytic leukemia (TLGLL) in Institute of Hematology & Blood Diseases Hospital in March 2020 were analyzed retrospectively, and the literatures were reviewed.Results:This male patient presented with recurrent infection when he was 17 years old, and was found neutropenia, anemia accompanied by obvious splenomegaly, lower level of IgG and IgA after the visit. The level of IgM was lower than the normal level and the typical XHIGM was manifested with the normal or increased level of IgM, however CD40L homozygous mutation (chromosome: chrX; location: 135730438; variation of amino acid: NM_000074:exon1:c.31C>T:p.R11X; nonsense mutation) was confirmed by next generation sequencing. CD40L heterozygous mutation was detected in his mother, but it was not in his father. The patient was diagnosed as XHIGM. Anemia and neutropenia were alleviated after splenectomy in the patient, who was diagnosed as T-cell large granular lymphocyte elevation and clonal proliferation by flow cytometry, TCR gene rearrangement positive and bone marrow histopathological immunohistochemistry results because of the increasing leukocyte. The patient was eventually diagnosed as XHIGM complicated with T-LGLL.Conclusions:A small number of patients with XHIGM may develop symptoms in adulthood and may present with atypical clinical features of significant reduction in IgG, IgA, and IgM. The confirmed diagnosis of XHIGM is established by identification of CD40L gene mutation. XHIGM gene screening is required in male patients with recurrent infection, IgG level lower than normal and neutropenia. A few XHIGM patients are complicated with T-LGLL.