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 investigate the relationship between the onset of lower extremity venous thrombotic disease and seasons.Methods A retrospective study was conducted, 300 patients with lower extremity venous thrombotic disease admitted to HandanCity Hospital of Traditional Chinese Medicine (TCM) from August 2012 to February 2014 were enrolled, the incidences and TCM syndromes of patients with lower extremity venous thrombotic disease in different seasons were observed, and the pathogenesis and relationships between the types of TCM syndrome and seasons were analyzed.Results There were 142 patients with lower extremity superficial thrombophlebitis, and 158 cases with lower extremity deep venous thrombosis, the incidence of lower extremity venous thrombotic disease in spring was significantly higher than that in summer and autumn [32.8% (86/262) vs. 21.3% (54/254), 18.4% (50/272), bothP < 0.01], but lower than that in winter [32.8% (86/262) vs. 37.2% (110/296)], the difference was not statistically significant (P > 0.01); while the incidence of lower extremity venous thrombotic disease in winter was significant higher than those in summer and autumn (allP < 0.01). The incidences of damp and heat downward flow type in autumn and summer were increased compared with those in winter and spring [55.6% (30/54), 60.0% (30/50) vs. 20.0% (22/110), 23.3% (20/86), allP < 0.01], while the incidences of lower extremity venous thrombotic disease with damp heat and stasis syndrome in winter and spring were increased compared with those in summer and autumn seasons [80.0% (88/110), 76.7% (66/86) vs. 44.4% (24/54), 40.0% (20/50)].Conclusions The incidence of lower extremity venous thrombotic disease is related to seasons, and the onset is high in winter and spring, damp heat and stasis syndrome being the main type; according to different seasons, clinical treatment can direct to different pathogenic factors to adopt different preventive measures interfering with the patient's constitution in order to eliminate or reduce the risk factors, achieving the effect of the disease prevention.

Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.

Objective To determine LDLR gene mutation in 2 clinically diagnosed FH patients from Hubei province and provide basis for gene diagnosis of FH.Methods Clinical data of 2 FH patients and their parents were collected.The promoter region and exon 1 to exon 18 region of LDLR gene were amplified through PCR and the amplified products were analyzed by forward and reverse DNA sequencing.The mutations were identified after comparison with LDLR gene sequence in GenBank.The pathogenic gene mutations were confirmed according to both genotype and phenotype of FH probands.Results The levels of plasma TC of two probands were 12.79 and 11.98 mmol/L.respectively.No gene mutations were detected in region 3 500 to 3 531 of ApoB100. The mutations of LDLR gene were compound heterozygous mutations. The novel mutation 665G > T detected in the exon 4 of No. 1 proband's LDLR gene was heterozygous missense mutation. The novel mutation 1 358 +32C > T was detected in the exon 9 of No. 1 proband's LDLR gene.The mutations 665G > T ( paternal origin) and 1 358 + 32C > T ( maternal origin) were inherited from the parents. A novel mutation 1 257 C > A was detected in the exon 9 of No. 2 proband's LDLR gene, resulting the presence of a premature termination codon, which was different from 1 257 C > G reported in Belgium.Another heterozygous missense mutation 1 879 G > A was detected in exon 13. They were derived from paternal origin and maternal origin, respectively. Conclusions There are three novel gene mutations:665G >T, 1 358 +32C > T, 1 257C > A found in two probands with compound heterozygous mutations in LDLR respectively. They maybe play a potential role in FH pathogensis.

ObjectiveTo investigate bronchial provocation test (BPT) and small airway function in children with cough variant asthma (CVA).MethodsA total of 353 children with chronic cough whose mean age was (7.45±2.58) years from three hospitals of Pudong district were enrolled during May 2012 and February 2014. Conventional pulmonary function tests, BPT and questionnaire survey were performed and the difference in pulmonary function was analyzed between children with positive BPT and negative BPT.ResultsIn 353 children with chronic cough, there were 200 children (56.66%) diagnosed as CVA with posi-tive BPT. Compared with BPT negative group, the percentages of nighttime cough and severe dry cough in BPT positive group were signiifcantly higher while the percentages of morning/daytime cough and wet cough were signiifcantly lower (P<0.01). Fur-thermore, the rates of history of atopic dermatitis and rhinitis in BPT positive group were signiifcantly higher than those in BPT negative group (P<0.01). Forced expiratory lfow at 75% relfecting the small airway function was signiifcantly lower in BPT posi-tive group than that in BPT negative group (P=0.032).ConclusionsBronchial hyperresponsiveness and decreased small airway function are the important pathological features of CVA. BPT and spirometry have clinical signiifcances in the CVA diagnosis and the analysis of cause of chronic cough.

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

Objective To discuss the treatment effect and application value of salmon calcitonin nasal spray in the patients with postmenopausal osteoporosis.Methods 130 patients with postmenopausal osteoporosis were divided into observation group and control group by random number method,65 cases in each group.The control group was given calcium and alfacalcidol treatment,the observation group received in combination with salmon calcitonin nasal spray treatment.The clinical effect of the two groups was recorded.Results The total effective rate of the treat-ment group was 95.38%,which of the control group was 84.62%,the difference between the two groups was statisti-cally significant (χ2 =4.188,P<0.05 ).After treatment,the tuberosity bone mineral density,femoral neck BMD, L1 -4 lumbar spine bone mineral density of the observation group were (0.69 ±0.15)g/cm2,(0.77 ±0.21)g/cm2, (0.98 ±0.26)g/cm2,which of the control group were (0.52 ±0.08)g/cm2,(0.64 ±0.13)g/cm2,(0.81 ± 0.15)g/cm2,the differences between the two groups were statistically significant (t=8.062,4.243,4.566,all P<0.05).After treatment,the calcitonin,beta collagen fragment,osteocalcin of the observation group were (2.97 ± 0.86)ng/L,(0.11 ±0.01)ng/mL,(7.18 ±1.14)ng/mL,which of the control group were (2.41 ±0.43)ng/L, (0.35 ±0.08)ng/mL,(15.89 ±3.53)ng/mL,the differences between the two groups were statistically significant (t=4.695,24.000,18.930,all P<0.05).Conclusion Salmon calcitonin nasal spray in the treatment of patients with postmenopausal osteoporosis can reduce pain,inhibit osteoclast activity,promote bone formation,prevent bone loss and increase bone mass,it is safe and worthy of popularizing in clinical application.

Objective To explore the correlations of diffusion-weighted imaging (DWI) types and the degree of neurologic impairment in acute ischemic stroke patients with atrial fibrillation.Methods DWI images and National Institutes of Health Stroke Scale(NIHSS) of 186 patients with acute ischemic stroke patients with atrial fibrillation were collected retrospectively.The correlation of DWI features and NIHSS was analyzed.Results On DWI,all acute ischemic stroke patients with atrial fibrillation presented high signal intensity.Single cortex-subcortical infarction mostly appeared in the anterior circulation(94,50.5%);Multi-infarction commonly occurred in the posterior circulation(18,13.0%);The neurological deficit scores of subcortical-cortex infarction in the left anterior circulation(16.75±7.10) were higher than that in the right side(13.50±5.70)(P<0.05).The neurological deficit scores of cortex-subcortical infarction in the posterior circulation (6.38±2.03) were significantly lower than that in the multi-infarction (16.77±8.90) (P<0.05).Conclusion DWI types are valuable for etiological diagnosis in ischemic stroke.Combination with NIHSS score could provide a basis for clinical individual treatment programs selection and prognostic evaluation.

Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.

Objective To screen monoclonal antibodies (mAbs) for early diagnosis of invisive Aspergillus. Methods Monoclonal antibodies against different antigens of Aspergillus fumigatus were produced. The two pairs of combinations of monoclonal antibodies were selected accoring the distinct epitopes and double-antibody sandwich ELISA based on mAbs above were established. The sensitivity and specificity of the methods were analyzed by detecting culture supernatants of clinical isolates and environmental isolatesof Aspergillus. spp, Penicillium Marneffei, Candidas, and serum from animal models and patients. The epitopes recognized by mAbs were identified by immunobotting. Results A total of 32 hybridoma cell lines that stably produced MAbs were obtained. Two double- antibody sandwich ELISAs were established. One method was specific for 19 clinical isolates and environmental isolates of Aspergillus. spp, whereas the other one was specific for the clinical and environmental isolates of Aspergillus fumigatus without cross-reation with other Aspergillus. spp. For the same kind of medium of Aspergillus fumigatus, the sensitivity of the first method was 10 fold higher than the second method. Conclusions The specific mAbs for early diagnosis of invisive Aspergillus were obtained. Antigen recognized by the specific mAbs was mannoprotein with molecular weights of approximately 25 000-75 000. This antigen was potential early diagnostic marker for invasive Aspergillus.