Objective To evaluate the diagnosis and treatment of primary breast lymphoma(PBL).Methods Clinical data of 49 PBL cases diagnosed in our center from 1997 to 2011 was retrospectively analyzed.Results 1 case was male,48 cases were female.Most patients were at the age of 35-55 years.Painless mass was the most common symptom which was most often seen on the upper-outer quadrant of the right breast,accounting for 61.11％ in this group.Preoperative misdiagnosis rate was high by mammography,B ultrasonic and hollow needle pathologic examination.Final diagnosis could only made by postoperative histopathologic examination.In this group 1 case was Hodgkin lymphoma and 48 cases were non-Hodgkin lymphoma(NHL) originating from B cells.46 patients underwent surgery and chemotherapy followed by radiotherapy in 16 cases.Median time of follow up was 40 months,5-year overall and disease free survival was 48％ and 28％ respectively,and bone marrow was the most common metastatic organ.Conclusions The prognosis of PBL is poor.The image diagnosis lacks specificity.Paraffin sections and immunohistochemistry were the main means to make a definite diagnosis.Chemotherapy-based comprehensive treatment is the strategy in the management of PBL instead of radical mastectomy.

Objective To explore the change of CD4+ CD25+ regulatory T cells(Treg) from peripheral blood in patiens with multiple organ dysfunction syndrome(MODS) before and after treatment and its significance.Methods The frequencies of CD4+ CD25+ CD127-Treg were detected in 10 patients with MODS respectively before treatment and 1 week after treatment and 10 healthy donors by flow cytometry labeled with specific fluorescent antibodies,such as anti-CD4(PE-CY5),anti-CD25(FITC) and anti-CD127(PE).Results The frequency of CD4+ CD25+ CD127-Treg in patients after treatment(2.11%?0.33%) was significantly lower than before treatment(7.44%?1.59%)(P

Objective To obtain the Hiwi gene encoding full-length and construct human Hiwi adenoviral vectors carrying green luorescence protein (GFP), and to establish foundation for a further study on Hiwi function and mechanism of inducing leukemia stem cell differentiation and apoptosis.Methods All coding areas of human Hiwi gene full length were amplified using method of overlapping extension PCR technology, and the full length coding aeras were inserted into the vector of Flag-IRES-hrGFP carrying GFP with Gateway technology to construct pDown-Hiwi-3 × flag-IRES-hrGFP. The cloning vector pDown-Hiwi-3 × flag-IRES-hrGFP and expression vector pAV. Des1d were used for homologous recombination reaction to obtain recombinant adenovirus vector pAV.Ex1d-Hiwi-3× flag-IRES-hrGFP.The positive clones were selected by PCR to extract the recombinant adenovirus plasmid and to pack into recombinant Ad-Hiwi-3 × flag-IRES-hrGFP adenovirus. Results The human recombinant Hiwi was successfully cloned and the recombinant adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was found to be successfully constructed via restriction enzyme digestion and sequencing methods. The adenovirus vector pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP was transfected into the HEK293A cells using lipofectamine mediated gene transfection method. Under fluorescence microscope, the transfected cells with green fluorescence could be observed.Conclusion The expression plasmid of adenovirus vector pAV.Ex1d-Hiwi-3 × flag-IRES-hrGFP is successfully constructed and it can express in HEK293A cells.

Objective:To clone human interleukin-33(hIL-33)and express it in E.coli efficiently.Methods:The primers were synthesized according to the hlL-33 cDNA sequence in GeneBank.The hIL-33 was amplified by RT-PCR from human fibroblast cell line(L929),the PCR product was inserted into pUC19 vector.The IL-33 cDNA confirmed by sequencing was inserted into expressing vector PQE30 and expressed in E.coli M15 strain.IL-33 protein expression was induced by IPTG and purified by Ni-NTA affinity chromatography.The recombinant IL-33 was identified by Immunoblot and its biological activity was analyzed.Results:DNA sequencing confirmed that the cloned cDNA was identical to the published sequence of hIL-33.The recombinant plasmid PQE/hIL-33 was transformed into M15.An expected 18KD protein of hIL-33 found mainly in the induced host strains about 25% of total bacteria lysis by SDS-PAGE and coomassie blue staining.The 18 KD protein could be recognized by anti-IL33 antibody in western blot.The recombinant protein was purified to more than 95% of total protein and induces the production of IL-4 and IL-5 in human peripheral blood mononuclear cells.Conclusion:We have successfully expressed hIL-33 protein in E.coli and the expressed product has IL-33 specific bioactivity.

ObjectiveTo analyze the Uridine diphosphoglucuronyl transferase 1 A1 gene (UGT1A1) mutation in non-he-molytic indirect hyperbilirubinemia.MethodsA female patient was diagnosed with type I Crigler-Najjar syndrome (CNS-I) after excluding hemolysis and hypothyroidism. Phototherapy treatment is effective while oral phenobarbital was not. UGT1A1 were ampliifed by polymerase chain reaction and DNA was sequenced.ResultsThe patient was compound heterozygote of c.1070A>G p. (Gln357Arg) and 1091C>T p. (pro364Leu) and diagnosed with CNS-I. Heterozygotes of the mutation were found in her parents.ConclusionsIn patients highly suspected of CNS, the genetic tests should be carried out as soon as possible.

Objective To identify the clinical characteristics and prognostic factors in young women of breast cancer treated with breast conservation therapy.Methods Clinical data of 605 cases of breast cancer patients undergoing breast conserving therapy were reviewed in our hospital from Jan 1998 to July 2008.According to their age,these patients were divided into young group (age≤ 35 years,n =110 cases) and the elderly group (age ＞ 35 years,n =495 cases).Survival factors were analyzed by Cox proportional hazards regression model.Results During the median follow-up of 61 months,the 5-year local-regional relapse-free survival was 92.6％ and 96.8％,respectively (P =0.074) ; the 5-year distant metastasis-free survival and overall survival between the two groups were (89.7％ vs.95.2％,P =0.001)and (96.4％ vs.98.9％,P =0.046).Multivariate analysis revealed that lymph node metastasis and adjuvant radiotherapy were independent risk factors for local-regional relapse.Distant metastasis and overall survival were determined by lymph node metastasis,adjuvant radiotherapy and age.Conclusions Age is not an independent factors affecting tumor local recurrence in Chinese young female patients of early breast cancer receiving breast conserving surgical therapy.

Objective To evaluated the effect of first-line epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI)on advanced non-small cell lung cancer (NSCLC)patients with different EGFR mutation status (exon 1 9 deletion and exon 21 mutation).Methods Seventy-two advanced NSCLC patients with EGFR mutation confirmed by histopathology were enrolled.All of the patients received first-line EGFR-TKI.The relationships between EGFR mutation status and objective response rate (ORR),disease control rate (DCR),progression free survival (PFS ) and overall survival (OS ) were analyzed.Results Of the 72 patients,37 patients expressed exon 1 9 deletion,35 patients expressed exon 21 mutation,and all of them could be evaluated.The ORR and DCR of patients with exon 1 9 deletion were higher than those of patients with exon 21 mutation (75.7%vs.51 .4%,χ2 =4.583,P=0.032;89.2%vs.68.6%,χ2 =4.636,P=0.031 ).The modified median PFS of patients with exon 1 9 deletion was significantly higher than that of patients with exon 21 mutation (1 3.2 month vs.1 0.8 month,χ2 =4.700,P=0.030).The median OS of patients with exon 1 9 deletion was significantly higher than that of patients with exon 21 mutation (30.2 month vs.25.6 month,χ2 =4.686,P=0.030).The side effects were similar between the two groups.The most common adverse reaction was rash,and the incidence had no significant difference between the two groups (48.7% vs.48.6%,χ2 =0.000,P=0.995 ).Conclusion EGFR mutation status is a predictor for PFS,OS and ORR of first-line EGFR-TKI in patients with advanced NSCLC.NSCLC patients with EGFR exon 1 9 deletion are associated with longer survival time and better response rate compared with those with exon 21 mutation.

Objective To measure and determine the position of the tibial attachment of the anterior cruciate ligament (ACL)in relation to its anterior ridge on the magnetic resonance imaging (MRI)of normal knee joints,and to explore the clinical role of the anterior ridge in guiding tunnel positioning during ACL reconstruction as well as in facilitating postoperative radiographic evaluation.Methods The knee MRI of one hundred young adult patients with an intact ACL and normal knee joint (mean age:25.1 years,range:18-40 years)was retrospectively reviewed.All MR images were obtained at full extension of the knee on the same MRI machine.Using digital image software on MRI,the measurements in the sagittal view were taken,including the depth of the tibia,the distance from the anterior edge of the tibial plateau to the most anterior and posterior portions of the ACL insertion on the tibia and the anterior ridge according to Staubli and Rauschning's method.Results The center of the tibial insertion of the ACL is located between 28.43％ and 50.94％ of the total anterior-posterior depth of the tibia,which was less than 43.3％ in 58 patients.The average distance from the anterior edge of the tibial plateau to the anterior ridge was 13.61 ± 2.17 mm (ranging from 8.03 to 18.65 mm),26.80％ ±3.89％ (ranging from 17.74％ to 33.94％)across the tibial plateau.There were significant positive correlations between the distance from the anterior edge of the tibial plateau to the most anterior portion of the ACL insertion and that to the anterior ridge.The distance from the most anterior portion of the ACL insertion to the anterior ridge was averaged 0.56 ± 0.68 mm (ranging from-0.28 to 2.71 mm).During the ACL reconstruction,with the anterior edge of the tibial tunnel determined at posterior 0.5 mm to the anterior ridge,the graft size as 8 mm,and the tibial guider angle set as 55 degree,96of the patients (96％)would have the center of the tibial tunnel located before the center of their native ACL attachment.Conclusions On sagittal MR images,the location of the anterior ridge and the most anterior portion of the ACL insertion correlated well,with the average distance between them of 0.56 mm.The study indicates that during ACL reconstruction,tibial tunnel drilling with the anterior edge of the ACL graft positioned at the anterior ridge can achieve a more anterior position than the traditional methods to orientate according to the center of the bone tunnel.

Objective To investigate the influences of 5-azacytidine (5-Aza) with different concentrations on mesenchymal stem cells (MSCs) proliferation and differentiation into myoblasts. Methods Bone marrow-derived MSCs of 4 weeks Wistar rats were separated and purified, and then treated with 5-Aza with different concentrations. The growth ability of cell was assayed with methyl thiazolyl tetrazolium (MTT) method. The expression of skeletal muscle actin in MSCs was determined with reverse transcription polymerase chain reaction (RT-PCR) and electrophoresis after MSCs were induced. Results The cell proliferation was not affected by 0 and 1 ?mol ? L-1 5-Aza . There were expressions of skeletal muscle actin treated with 3 -12 ?mol ? L-1 5-Aza. Some cells were obviously enlarged at the 9th day after induction and myotubu-like cells were found at the 12th day when treated with 9 and 12 ?mol ? L-1 5-Aza. 20 - 30 ?mol ? L-1 5-Aza induced the toxic effect on proliferation of MSCs. With the increase of concentration, the proliferation ability of MSCs was weakened. Conclusion 5-Aza affects the expression of regulatory gene to the stem cells and regulate MSCs to orientationally differentiate into myotube-like cells.

Objective To analyze the gene abnormality and liver pathology in Shwachman-Diamond syndrome (SDS) in a child. Method The clinical data of one child with SDS were analyzed retrospectively. Result A male patient was 1 month old at onset with neutrophil decrease as the first manifestation, accompanied by anemia, elevated transaminase and repeated infection. Exocrine pancreatic dysfunction was atypical. Pathological examination of liver biopsy showed slight damage of liver cells under light microscope. The blood samples of child and parents were collected, and homozygous mutations of SBDS (NM_016038.2) Intron2 c.258+2T>C p.? were detected by two generation gene sequencing. And these mutations were from both his parents. Conclusion Gene testing is helpful in diagnosing SDS and we suggest that liver biopsy should be performed if condition allows.