In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

OBJECTIVETo explore the expression of MICA/B in human esophageal cancer, and to analyze its correlation with clinicopathological features.

METHODSThe expression of MICA/B in 40 cases of esophagus carcinoma and corresponding normal esophageal mucosa tissues were examined by immunohistochemistry.

RESULTSThe positive rate of expression of MICA/B protein in the esophageal carcinoma was 75.0% (30/40), and that in the corresponding normal esophageal mucosa was 0 (0/40). Up-regulation of MICA/B expression was found in the esophageal carcinomas. The expression of MICA/B was related with histological grade of the esophageal carcinoma (P = 0.012).

CONCLUSIONMICA/B protein plays an important role in the esophageal carcinogenesis, and my become a useful molecular marker for the diagnosis of esophageal carcinoma.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Esophageal Neoplasms ; diagnosis ; metabolism ; pathology ; Female ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Grading ; Up-Regulation

Objective To explore psychological stress factors of patients in the ophthalmology department and psychological nursing interventions as well as effect, to promote the rehabilitation of patients. Methods A total of 289 patients from our ophthalmology department was investigated with the self-designed questionnaire. Results Major psychological stress factors of Ophthalmology inpatient were knowing nothing a variety of points for attention(92％), fearing and worrying the disease and surgery(85％), worrying about the level of health care technology of medical staff (84％). Psychological stress of patients released through psychological nursing intervention measures such as cognitive intervention and environmental intervention. 243 patients of fearing and worrying the disease and surgery reduced to 35 before and after intervene, The difference was statistically significant (X2 = 299.84, P ＜ 0. 01). Conclusions We should properly guide and inspire positive mental patients based on patients' mental state, give mental nursing intervention, promote the rehabilitation of eye disease.

To explore differentially expressed genes in leukemia gene expression profile and identify main related genes in acute leukemia, gene expression profiles were analyzed in bone marrow/leucopheresis peripheral blood stem cells samples from 9 acute leukemia patients and their sibling donors with the use of oligonucleotide microarrays. 163 reported leukemia-related genes were involved in the study. The oligonucleotide primers were designed, synthesized and spotted on the chemical-material-coated-glass plates in array. The total RNAs were isolated from nine patients' bone marrow or leucopheresis peripheral blood cells and from nine their sibling donors peripheral blood stem cells treated by G-CSF, then collected by CS-3000 cell selection machine, and were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the oligonucleotide microarray. The results showed that in four patient/donor pairs with B-ALL, 5 up-regulated (RIZ, STK-1, T-cell leukemia/lymphoma 1A, Cbp/p300, Op18) and 1 down-regulated genes (hematopoietic proteoglycan core protein) were identified; In five patient/donor pairs with AML-M(4) and AML-M(5), 6 up-regulated (STAT5B, ligand p62 for the Lck SH2, CST3, LTC4S, myeloid leukemia factor 2 and epb72) and 1 down-regulated genes (CCR5) were identified. In conclusion, on the basis of distinguishing study of specific genetic related recipient/sibling donor pairs, screening leukemia-related genes with oligonucleotide microarrays, a set of 13 up-regulated or down-regulated genes among 163 leukemia-related genes has been identified. The result has further confirmed that above genes play critical role in the molecular mechanism of acute leukemia.
Blood Donors ; DNA-Binding Proteins ; genetics ; Gene Expression Profiling ; Glutathione Transferase ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Microtubule Proteins ; genetics ; Milk Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; Peripheral Blood Stem Cell Transplantation ; Phosphoproteins ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; STAT5 Transcription Factor ; Siblings ; Stathmin ; Trans-Activators ; genetics

This study was aimed to investigate the expression levels of CXCR4 and VEGF in serum of patients with DLBCL and their clinical significances. The peripheral blood of 44 patients with newly diagnosed DLBCL and 20 healthy adults as a control group were chosen for study. And the expression levels of CXCR4 and VEGF in serum were detected by ELISA. The results showed that the expressions of VEGF and CXCR4 in DLBCL patients were higher than those in the control group (P < 0.05). The expression of VEGF was positively correlated with the expression of CXCR4 in DLBCL patients, and the correlation coefficient was 0.743 (P < 0.05). The VEGF expression in DLBCL patients was correlated with LDH, immunotyping, the number of extranodal involvements, Ann Arbor staging and ECOG performance score; while the expression of CXCR4 was correlated with LDH, immunotyping, the number of extranodal involvements and Ann Arbor staging. Univariate analysis showed that LDH, extranodal involvements, immunotyping, Ann Arbor staging, CXCR4 and VEGF were associated with OS. Multivariate analysis showed that the immunotyping and CXCR4 expression independently associated with OS. It is concluded that both expression levels of VEGF and CXCR4 are significant higher than those in the control group. CXCR4 expression positively correlates with VEGF expression and displays a prognostic significance for OS. This study suggests that combined targeting VEGF and CXCR4 may become a novel therapeutic strategy for diffuse large B cell lymphoma.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Receptors, CXCR4 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

Objective To investigate the safety and efficacy of re-irradiation with stereotactic body radiotherapy(SBRT) for treating locally recurrent advanced pancreatic cancer.Methods From 2014 to 2017,7 patients with stage Ⅲ pancreatic cancer were treated by re-irradiated with SBRT at Shanghai Changhai Hospital.SBRT was delivered via the G4 type cyberknife robotic stereotactic radiosurgery system in all the patients.The median dose of the first SBRT was 35Gy/5-7 fx,and the median dose of re-irradiation with SBRT was 31 Gy/5-8 fx.6 patients had undergone sequential chemotherapy either with gemcitabine or S-1 based therapy except one patient who refused the chemotherapy.Results There were 5 male and 2 female patients.The median overall survival (OS) of 7 patients was 30 months.Patients were re-irradiated with SBRT after a median interval of 10 months after the first SBRT.Median OS and locally relapse-free survival (LFRS) from re-irradiation were 13 months and 11 months,respectively.Three months after re-irradiation,3(42.9％) patients had partial remission and 4 patients had stable disease.Pain disappeared in 4 patients at the end of reirradiation and significant pain was alleviated in 2 patients 1 month after re-irradiation.There were no toxicities of grade 3 or higher grade during two courses of SBRT.Conclusions For patient with locally recurrent advanced pancreatic cancer,SBRT re irradiation regimen was associated with acceptable toxicity,which can effectively alleviate the pain,prolong the survival and improve the life quality.

Objective To compare the effect of staple food containing edible medicinal foods versus wheat fiber on blood glucose control in patients with type 2 diabetes mellitus.Methods We recruited 120 patients with type 2 diabetes mellitus into this study, and randomly divided them into 2 groups ( 80 participants in the study group and 40 in the control group) .The staple food in the study group was steamed bread with wheat flour and flour of several kinds of edible medicinal foods, and in the control was steamed bread with wheat flour and wheat fiber.The study lasted for 13 weeks.The changes of anthropometry, biochemical tests, blood glucose and insulin levels before and after the dietary intervention were compared between the two groups.Results Com-pared with the control group, patients in the study group showed more remarkable improvement in diastolic blood pressure (-0.05 mm Hg vs.0 mm Hg, P=0.021 ) , triglyceride level ( -0.10 mmol/L vs.0.27 mmol/L, P=0.040) , fasting and 2-hour postprandial blood insulin levels ( -1.30 mU/L vs.0.62 mU/L, P=0.001;-14.82 mU/L vs.-3.93 mU/L, P=0.020) .The patients in the control group had a larger reduction in waist circumference compared with the study group (-5.00 cm vs.-4.00 cm, P=0.001).There was no significant inter-group difference in changes of body weight, hip circumference, systolic blood pressure, or glucose tolerance ( P>0.05) .Conclusions Compared with wheat fiber, staple food containing wheat flour and edible medicinal foods can reduce triglyceride and insulin levels more significantly in patients of type 2 diabetes mellitus, while staple food supplemented with wheat fiber has a more obvious effect on waist circumference.

OBJECTIVETo study the function of the SP3111 protein in fertilization and early embryo development through in vitro fertilization (IVF) experiments following anti-SP111 antibody (Ab2438) blocking.

METHODSSperm samples collected from male mice were divided into an experimental, a blank control and a negative control group before IVF. The sperm of the experimental group was incubated with Ab2438 for 1 h followed by IVF and observed for the rates of fertilization and embryo fragmentation at 2, 4, 6, 8 and 22 h. Then the fertilized eggs were incubated with Ab2438, and the rates of fertilization embryo fragmentation were observed at 22 h.

RESULTSAfter the sperm was incubated with Ab2438, the incidences of embryo fragmentation were 5.26, 8.77, 23.25, 43.42 and 59.21% at 2, 4, 6, 8 and 22 h, respectively, with significant differences from the control groups (P < 0.01). After 22 h Ab2438 incubation of the fertilized eggs, the rates of normal and fragmented embryos of the experimental group were 23.64 and 63.64%, respectively, significantly different from those of the control groups (P < 0.01).

CONCLUSIONAnti-SP3111 antibodies remarkably affected fertilization and early embryo development in mice. The SP3111 protein may be a signal molecule and plays a role in fertilization and early embryo development together with other proteins. Further studies on the function of the SP3111 protein in reproduction may offer a new insight into the molecular mechanism of infertility.

Animals ; Antibodies ; immunology ; Embryonic Development ; Female ; Fertilization in Vitro ; Male ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred ICR ; Spermatozoa ; immunology

BACKGROUNDDifferent kinds of minimally invasive surgery (MIS) procedures have now been used in total knee arthroplasty (TKA). Compared with traditional TKA procedure with a long skin incision, clinical studies showed MIS procedures had some advantages. Quadriceps sparing (QS) procedures are the most minimally invasive MIS procedure until now. This study was aimed to find the insertion types for Chinese patients' vastus medialis and if the QS procedure had some advantages in patients' early recovery.

METHODSBetween February 2006 and May 2007, 120 consecutive patients underwent unilateral primary TKA under general anesthesia, among whom 14 patients were lost to follow-up, the remaining 106 cases were enrolled in this study. Among the 106 cases there were 85 right knees, 21 left knees (15 men and 91 women, with a mean age of 65.1+/-7.4 years); osteoarthritis in 97 patients (91.5%) and rheumatoid arthritis in 9 patients (8.5%). MIS TKA was performed in 49 cases (MIS TKA group), while MIS-QS TKA in 57 cases (MIS-QS TKA group). During the operation, the type I, II and III insertions of the vastus medialis for all patients were recorded. Each knee was rated post-operatively according to the Hospital of Special Surgery (HSS) scoring system. Clinical follow-up was undertaken at 1 week, 2, 6, 12 and 24 weeks. Operating time and complications were recorded.

RESULTSThere was no statistically significant difference between the two groups for gender distribution, age, left or right knee incidence, pre-operative diagnosis, incidence of varus or valgus deformity. Of the 106 cases there was 1 (0.9%) case with a type I insertion of the vastus medialis, 4 (3.8%) cases with type II insertions, 101 (95.3%) cases with type III insertions. The HSS scoring was significantly different between the MIS-QS TKA group and MIS TKA group within the first two weeks post operation. From 2 weeks later to 24 weeks, no significant difference was found. The average operating time was (53.3+/-12.4) minutes in the MIS TKA group and (64.1+/-15.1) minutes in the MIS-QS TKA group (P<0.001). In the MIS-QS TKA group, 1 patient had delayed healing of the partial skin incision (1.8%). No other complications were found in either group.

CONCLUSIONSAlthough most of the Chinese patients had type III insertions of the vastus medialis, the MIS-QS TKA procedure showed less injury to the quadriceps than the standard MIS TKA and this could contribute to the earlier recovery of the patients. But a shorter skin incision and more tension on the skin may also lead to more skin complications.

Aged ; Aged, 80 and over ; Anesthesia Recovery Period ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Quadriceps Muscle ; surgery ; Time Factors

BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.

METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.

RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.

CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult