The mutation status of calreticulin gene (CALR) is helpful for the diagnosis of JAK2 / MPL mutation-negative myeloproliferative neoplasms (MPN), and is closely related to the MPN progression and prognosis.Currently, Sanger sequencing, PCR-fragment analysis, high resolution melting, Taqman RQPCRand the next generation sequencing have been reported to be used to detecting the CALR gene mutations.A proper method for CALR mutation detection and a right laboratory diagnostic procedure according to the MPN-related molecular markers will facilitate the rapid and effective diagnosis and treatment of MPN.

Myelodysplastic syndromes ( MDS ) represent a heterogeneous group of clonal hematopoietic disorders with diverse clinical course .Because of the heterogeneity and the lack of molecular markers to monitor disease progression, clinical management of MDS patients is challenging .Recently, with the development of molecular analysis techniques , an increasing number of MDS related molecular biomarkers have been reported.In this review, we will discuss the clinical applications of the newly reported molecular makers in terms of diagnosis , prognosis and treatment.These markers may improve the diagnosis and prognostic assessment systems of MDS , which may potentially be used to guide decision making in the individual therapy.

Objective To study new active constituents in the mature fruits of Momordica charantia.Methods M.charantia was extracted by alcohol and purified by D-101 macroporous adsorptive resin.The isolation and purification were carried out by silica gel and semiprepared HPLC,the compounds were identified and elucidated by spectral and chemical methods.Results Eight compounds were obtained and identified as charantadiol A(5?,19-epoxycucurbita-6,23(E),25(26)-triene-3?,19(R)-diol,Ⅰ),(+)-eduesmin(Ⅱ),bluemenol A(Ⅲ),karatavilagenin D(Ⅳ),5?,19-epoxycucurbita-6,23-diene-3?-19,25-triol(Ⅴ),5,19-epoxy-5?-cucurbita-6,23(E)-diene3?,25-diol(Ⅵ),3?,7?-trihydroxy-cucurbita-5,23(E)-diene-19-al(Ⅶ) and ?-sitosterol(Ⅷ).Conclusion Compound Ⅰ is a novel one,named charantadiol A.Compounds Ⅱ and Ⅲ are obtained from M.charantia for the first time.

Objective To apply the microbial transformation to transforming the total saponins in the fruit of Panax ginseng (SFPG) and preparing ginsenoside compound K (C-K). Methods The four microbial strains m14, m3, m8, and m9 were screened and isolated from the soil in the botanic garden planted for P. ginseng and they were used for the microbial transformation of SFPG to optimize the strains. Taking C-K contents as index the microbial transformation was detected and analyzed by TLC and HPLC. Results The strain m14 was found to transform the SFPG efficiently to C-K at first. The optimal culturing and transformation conditions of m14 were obtained: time, 6 d; temperature, 30 ℃; revolution of cradle, 160 r/min; initial pH value, 5.5; substratum concentration, 120 mg/mL. Under the optimal condition, the content of C-K was 41.65 times as much as before transformation by m14. Conclusion The m14 is the most effective strain among the four fungal strains. It is the new way available for the C-K industrialized production.

Objective To improve the clinical and imaging diagnosis of benign fibrous histiocytoma of bone.Methods Imaging findingsin 14 patients with pathologically confirmed benign fibrous histiocytoma of bone were analyzed.X-ray plain film was performed in allpatients,CT scan and MR scan were done in 9 cases and in 5 cases respectively.Results The clinical symptom was mainly pain in the local lesion,soft tissue mass could be felt around the lesion in 2 cases.The single-lesion was 12 cases and multi-lesion was 2 cases.There were 22 lesions in 14 cases.Most lesions localized in long bones,totally were 17 lesions.9 lesions localized in cancellous bone,8 lesions localizedin diaphysis compact substance.X-ray plain film showed centric or eccentric osteolytic destruction,unilocular or multiocular with clearborder or sclerotic border,expansion,pathologic fracture and speckled calcification in some destruction lesions.Soft tissue mass could be seen around the lesion in few cases.CT was better than X-ray plain film in displaying calcification,cyst change,pathologic fracture and soft tissue masses in the destruction portion.MR presented as homogeneous signal on T_1WI and hyper-signal on T_2WI,sometimes was inhomogeneous,speckle-like low-signal and non-signal could be seen in the lesion on T_1WI and T_2WI.Few lesion shows intermixed signal on T_1WI and T_2WI.Conclusion Based on the typical imaging and clinical features,the correct diagnosis of benign fibrous histiocytoma can be made in most cases.

Objective:To study the regulatory effects of Ligustrazine Hydrochloride(LHC)on tumor-induced immunosuppression by Colon26 cells in vitro.Methods:Colon26 cells were cultured for 48 h in the presense of LHC and either the cell fraction or the cultural supernatants was collected,with the untreated Colon26 cells as control for the study.The down-regulating effects of LHC on tumor immunosuppressions (including the suppressed NK killing and ConA induced transformation of murine spleen cells detected by MTT,and the reduced expression levels of IL-2R?,CD3?+?+ and CD3?-?+ detected by FCM) were determined.The concentrations of immunosuppressive cytokines,including TGF-?1,VEGF,IL-4,IL-6 and IL-10,in the supernatants were analyzed by quantitative ELISA.The relationship among the down-regulatory effects of LHC on secretion immunosuppressive cytokines and tumor immunosuppressions were evaluated by multiple linear regression analysis.Results:All of the cytokines assayed were found in the supernatant of Colon26 treated without LHC,in which TGF-?1 was the highest,and the significant inhibition of five immune functions mentioned above was showed.To the Colon26 treated by LHC,the concentrations of TGF-?1,IL-6 and IL-10 in the first re-cultured supernatant and its inhibition of five immunol functions decreased greatly.The concentrations of TGF-?1 and IL-6 in the second re-cultured supernatant and its inhibitions of transformation,CD3+?+ and CD3-?+resumed highly.The positive correlations existed between TGF-?1 and inhibition of immunol functions except for transformation,between IL-6 and inhibition of transformation or CD3-?+,between IL-10 and inhibition of NK killing or IL-2R? or CD3+?+,respectively.Conclusion:LHC can exert down-regulate effects on Colon26 secretion of immunosuppressors and its tumor immunosuppression.Reducing tumor immunosuppression of Colon26 through decreasing its secretion of immunosuppressors should be one of anti-tumor mechanisms of LHC.

Objective:To obtain the prokaryotic expression product for the group 6 allergen of Dermatophagoides farine ( Der f 6) and detect its IgE-binding rates with sera from asthmatic children. Methods: By enzyme digestion of pET28a (+)-Der f 6 with BamHⅠ plus XhoⅠ,the target gene Der f 6 was obtained and linked into the vector pET32a (+) to construct the recombinant plasmid pET32a(+)-Der f 6, which was then transfected into E. coli BL21 cells for expression, induced with isopropyl-β-D-thiogalactoside ( IPTG) ,purified by affinity chromatography and identified by SDS-PAGE,Western blot and AMLDI-TOF,and tested by ELISA for IgE reactivity with sera from asthmatic children. Results:The plasmids pET32a(+)-Der f 6 were constructed,transformed into E. coli BL21 and expressed successfully. SDS-PAGE of the purification product showed a specific band,Western blot showed the successful binding between the purification product and the His-tag in the plasmids,and MALDI-TOF/TOF identified the identical structure to the allergen Der f 6. Using the ELISA method developed with the recombinant proteins as coating antigen,the positive rate was 41. 3% (19/46) in asthmatic children allergic to dust mite. Conclusion: The plasmids pET32a (+)-Der f 6 were constructed successfully,expressed in E. coli BL 21 (DE3). The recombinant fusion protein has a good reactivity with sera from asthmatic children.

Objective To assess early and medium outcomes of pathologic N2 disease unexpectedly detected in patients undergoing total video-assisted thoracic surgery lobectomy for non-small cell lung cancer.Methods Between Sep.2006 and Dec.2010,348 patients with Non-small cell lung cancer underwent total video-assisted thoracic surgery lobectomy,and within them,35( 10.1％ ) were found to have pathologic N2 disease after operation.We retrospectively reviewed the clinical and pathologic features of patients with unexpected N2 disease after video-assisted thoracic surgery lobectomy and their early and medium outcomes,including survival and recurrence pattern.Results No perioperative mortality was noted.26 patients received a lobectomy directly,and the other 9 patients after a wedge resection.All the patients had R0 resection.The medium operation time was 190 minutes and medium blood loss was 200ml.The medium stations and numbers of dissected N2 lymph nodes in operation were 4 and 10,respectively.And the medium stations and numbers of metastatic N2 Lymph nodes were 1 and 2,respectively.Among patients with pathologic N2 disease,18 (51.4％) had single-station involvement.The median duration of chest tube placement was 8 days.The median length of hospital stay was 11 days.15 complications occurred in 12 (34.3％) patients.All of the patients underwent adjuvant chemotherapy with platinum postoperatively.The median follow-up time was 23 months.The 1 - and 2-year overall survival (OS) was 80.9％ and 67.9％,and the medium OS was not reached.During follow-up,16 (45.7％) patients had a recurrence.The pattern of recurrence was locoregional in 5,distant in 11.The 1 - and 2-year disease-free survival (DFS) was 71.9％ and 44.2％,and the medium DFS was 20 months (95％,8.1 to 31.9 months).Divided the patients with pathologic N2 disease into two groups considering single-station involvement or not,the 1-and 2-year OS and DFS for the single-station group and for the multiple-station group were 87.7％,78.9％ ; 88.9％,49.4％and 67.6％,59.1％ ; 55.3％,39.5％.The medium DFS for both the two groups was 23 and 16 months respectively.Conclusion For non-small cell lung cancer with N0 disease confirmed by an exactly preoperative staging workups,if it is feasible in technology,a total video-assisted thoracic surgery lobectomy should be recommended.Even if N2 lymph node metastasis is unexpectedly detected postoperatively,the metastasis was mostly micro- or single-station involved,and a similar outcome with conventional thoracotomy can be achieved.

OBJECTIVETo develop an HPLC for determination of aglycone of momordicoside L in Momordica charantia.

METHODA Kromasil C18 (4.6 mm x 150 mm, 5 microm) column was used. The mobile phase was acetonitrile-H2O (64:36), the flow rate was 1.0 mL x min(-1) and the UV detection wavelength was set 203 nm.

RESULTThe calibration curves were linear from of 0.025 microg to 1 microg (r =0.9911), the contents of aglycone of momordicoside L in Shandong, Henan, Hebei, Jiangxi are 0.211, 0.033, 0.013, 0.007 mg x g(-1), respectively.

CONCLUSIONThe method is simple and reliable for determination of aglycone of momordicoside L in M. charantia.

BACKGROUND:Pedicle screw is the major instrumentation of surgery in thoracic spine. However, there have been few reports about pedicle morphology relevant to screw insertion tracts, and few reports comparing the normal adolescents and adolescent idiopathic scoliosis patients. OBJECTIVE:To compare the morphologic characteristics of the thoracic pedicle with regard to safe thoracic pedicle screw placement in normal adolescents and adolescent idiopathic scoliosis patients. METHODS: Thoracic pedicles of thirty-five normal adolescents and thirty-five adolescent idiopathic scoliosis patients were measured with three-dimensional reconstruction CT images. Measured parameters include (1) critical distance: the shortest distance from an entry point to the ventral cortex of the lamina. (2) Safe distance: the distance from the entry point to the tangent of the spinal canal at the medial wal of the pedicle. (3) Pedicle screw length. (4) Pedicle width. (5) Pedicle transverse angle. The dangerous area was defined as the distance between the critical distance and the safe distance. RESULTS AND CONCLUSION: The mean critical distance was (9.2±1.0) mm for the normal adolescents, and (9.4±1.2) mm for the adolescent idiopathic scoliosis patients. Safe distances were significantly less in normal adolescents (14.7±0.8) mm than that of the adolescent idiopathic scoliosis group (15.4±1.4) mm (P < 0.001). The dangerous area was (5.4±0.7) mm for the normal adolescents, which was significantly less than that of the adolescent idiopathic scoliosis patients (6.0±1.0) mm (P < 0.001). Pedicle screw length was (36.6±4.1) mm for the normal adolescents and (37.1±5.3) mm for the adolescent idiopathic scoliosis patients. Pedicle width was (5.8±1.2) mm for the normal adolescents and (5.7±1.7) mm for the adolescent idiopathic scoliosis patients. No significant difference in critical distance, pedicle screw length and pedicle width was found between the two groups (P=0.382, 0.135, 0.293). Pedicle transverse angle decreased gradualy from T1 to T12 in both groups. These results verify that pedicle morphology of many parameters is different between normal adolescents and adolescent idiopathic scoliosis patients, especialy in the apical area of the thoracic curve.