Objective To evaluate the value and significance of testing megakaryocyte micronuclei in bone marrow smears for hematopathy diagnosis.Methods Bone marrow smears from a total of 863 cases of patients with hemopathy were collected from 2002 to 2009 at the second affiliated hospital of zhejiang university school of medicine.Smears from 25 healthy individuals were used as control.The bone marrow smears were subjected to Wright-Giemsa staining.The number of megakaryocytes, morphous and positive rate of megakaryocyte micronucleis were recorded by using low power lens and immersion objective.Statistical differences of positive rate in the different diseases, pathologic subtypes, total number and type of megakaryocyte were analyzed.Results The megakaryocyte micronucleis were round or oval with varying size, distributed in paranuclear or away from nuclear and even free in extracellular, which could be observed in large and medium megakaryocytes.The positive rate of megakaryocyte micronucleis was highest in myeloid leukemia, particularly in the subtype of M6, M2, M4 and M5b with dyshaematopoiesis.Megakaryocyte micronucleis could also be found in MA, infection and benzolism, but rarely observed in the lymphocytic leukemia.Conclution The detection of megakaryocyte micronuclei was related with its amount, size and type.There is significant difference of the postive rates of megakaryocyte micronuclei testing among the different hematopathies and pathologic subtypes.Megakaryocyte micronuclei testing should be valuable in the the diagnoses of hematopathy.

Objective To study the relationship between the fbxl 20 gene、E-cadherin gene and the clinocopathologic features ,respectively , and the correlation about the fbxl 20 and the set gene .Methods The mRNA expression level of fbxl 20 and E-cadherin gene in 50 pairs of human colorectal adenocarcinoma tissues and matched normal tissues were detected by RT -PCR.Results The mRNA expression level of fbxl 20 gene in tumor tissues were significantly increased than that in normal tissues (0.479 ±0.141 vs.0.296 ±0.121,P=0.001),while the E-cadherin were decreased (0.440 ±0.026 vs.0.741 ±0.059,P=0.000).There was no found on the correlation between the mRNA expression level of fbxl 20 and the E-cadherin gene in the 50 tumor tissues(r=-0.165,P=0.251).However,the E-cadherin expressed level was negatively correlated with the fbxl 20 expressed level in the lower differentiation degree、in the high differentiation degree and the Duke′s D tumor tissues(r=-0.600,P=0.008;r=-0.784,P=0.017;r=-0.643,P=0.032).Conclusions The decreased mRNA expression level of E -cadherin in tumor tissues are largely due to the increased fbxl 20 gene expressed level ,which is related with the high mobility and invasion ability of the advanced tumor .

Objective To observe the inhibitory effect of DPC4 gene on growth of pancreatic carcinoma in vitro. Methods The human DPC4 cDNA was subcloned to the retroviral vector pLXSN and then packaged with GP+E86 and PA317 packaging cells respectively. AntiG418 clones were acquired and named as PA317/ pLXSN DPC4+ cells. The DPC4 gene was restored to the human pancreatic cancer cell line BxPC-3 by the infection of the pLXSN/DPC4 and then had a stable expression after antiG418 selection, which was demonstrated by RT-PCR and Western blot. The inhibitory action of DPC4 gene expression on growth of these daughter cells was observed.Results DPC4/Smad4 gene integration in GP+E86、PA317/ pLXSN DPC4+ cells was detected by polymerase chain reaction. The BxPC-3 cells, which were null for DPC4, had stable expression of DPC4 after the infection of the pLXSN/DPC4. The expression of DPC4 gene was able to inhibit the growth of these cancer cells in vitro and downregulate the VEGF mRNA level.Conclusions This study suggested that there can be marked inhibition of growth and angiogenetic ability of pancreatic cancer cells after infection by retrovirus vector containing DPC4.

Objective To explore the diagnosis value of morphology changes of pleomorphic megakaryocytes in the bone marrow (BM) smears and BM sections in chronic MPD(CML-CP, ET,PV and PMF). Methods BM aspiration was taken in 182 patients of MPD aspiration and biopsy examination was performed synchronously to obtain the BM smears and BM sections samples. The BM smears were subjected to Wright/Giemsa stain and immunohistochemistry stain, while the BM sections were subjected to Haematoxylin-Giemsa-Fuchsin stain. The morphology of pleomorphic megakaryocytes was classified into five groups, which were Ⅰ type ( inclusion type), Ⅱtype ( hypolobulated muclei type), Ⅲ type ( giant hyperlobulated nuclei type), IV type (micro pyknotic type), and V type(extrusion type). The size of megakaryocytes clusters was recorded as no clusters(0) , predominantly small clusters of fewer than 6 cells (1) or predominantly large clusters of at least 6 cells (2) . The detection rates of various types of pleomorphic megakaryocytes and megakaryocytes clusters were both analyzed in the BM smears and BM sections. Results In CML-CP group, the detection rates were (3. 73±3. 84)% , (14.19 ±7. 62)% ,(5.99 ±4.67)%, (34. 37 ±10.79)%, (9.45 ±6. 87)%, (32. 28 ±7. 67)% and 3.13 ±2. 30)% ,(12.61 ± 9.28)%,(4.94±4.27)%,(35.26±9.63)%,(9.47 ±5.89)%,(34.58 ±6.81)% for I tⅠype,Ⅱ type,Ⅲ type, Ⅳtype and Ⅴ type pleomorphic megakaryocyte in BM smears and BM sections. There were no significantly differences between the BM smears and BM sections(t value were 0.524,0.510,0.645, 0.239,0.011,0. 869,all P>0.05). In ET group, the detection rate of I type [ (6.17 ±2. 89)% ] in BM smears was significantly higher than that in BM sections [ 2.42 ± 1. 28) % ] (t = 7. 183, P < 0. 01) , while the detection rate of V type [ (6. 28 ± 3. 34) % ] in BM smears was significantly lower than that in BM sections [ (10. 18± 4.03) % ] (t = 3.940, P < 0.01). Besides these, the detection rates of other types were not significantly different between the BM smears and BM sections(t value were 0.079,0. 122,1.643, 1. 638,all P>0. 05). In PV group, the detection rate of V type in BM smears [ (6. 55 ±4. 11)% ] was significantly lower than that in BM sections [ (10. 30±3. 34) % ] (t = 2. 351, P < 0.05 ). However, the detection rates of the other types were not significantly different between the BM smears and BM sections (t value were 1. 635,0. 301,0. 132,0. 704,0. 681 ,all P' >0. 05). In PMF group, the detection rate of IV type in BM smears [(13.05 ±5.24)%] was significantly lower than that in BM sections [(29.14± 8. 72) % ] (t = 5. 245, P < 0. 01). And the detection rate of normal type in BM smears [ ( 33. 58 ± 14.39)% ] was significantly higher than that in BM sections [(23. 01±7.96)%] (t =2. 132,P<0. 05). Besides these, the detection rates of the other types were not significantly different between BM smears and BM sections( t value were 0. 787,0.646,2.062,0. 869, P > 0. 05 ) . In CML-CP and PV groups, the detection rates of size of clusters were not significantly different between the BM smears and BM sections (x~2 = 2. 772, P > 0. 05 ). In ET group, the detection rate of small clusters (1) in BM smears was obviously higher than that in BM sections, however, the detection rate of larger clusters (2) in BM smears was obviously lower than that in BM sections (x~2 = 13. 748, P < 0.01). In PMF group, the detection rate of no clusters(0) in BM smears was obviously higher than that in BM sections, however, the detection rate of large clusters(2) in BM smears was obviously lowers than that in BM sections (x~2 =18.741 ,P<0. 01). Conclusions Both BM smears and BM sections can be applied to observe pleomorphic megakaryocytes. The morphology changes of pleomorphic megakaryocytes have certain reference values for identification of MPD subtypes and differential diagnosis.

Objective To study the influence of CYP3A5 genetic polymorphism on tacrolimus metabolism in renal transplantation recipients and evaluate the methods of detecting CYP3A5* 3 polymorphism. Methods Ninety-seven renal recipients receiving the triple immunosuppressive (tacrolimus + mycophenolate mofetil + prednison) were genotyped for CYP3A5* 3 polymorphisms by the Pyrosequencing TM assays. Tacrolimus trough concentration of the patients was measured by enzyme multiplied immunoassay technique, and concentration/adjusted dose ratio (C/D) was investigated at the 7th day, 14th day, 1st month, 3rd month and 6th month after renal transplantation. Results The Pyrosequencing TM assays allowed for quick and accurate detection of CYP3A5* 3 genotypes. There were CYP3A5* 1/* 1 genotype in 17 cases (17. 5%), * 1/* 3 in 35cases (36. 1 %) and *3/* 3 in 45 cases (46.4 %) identified in 97 renal recipients. The C/D of CYP3A5 * 1/* 1 and * 1/* 3 patients was significantly lower than that of * 3/* 3 patients (P<0. 01)after renal transplantation. Conclusion The CYP3A5* 3 polymorphisms are significantly correlated with tacrolimus pharmacokinetic in renal transplant recipients. Detecting the CYP3AS* 3 genotype of the recipient before the transplantation by the Pyrosequencing TM assays can be used to help determine the optimal initial dosage after transplantation, resulting in individualized drug therapy.

Objective To investigate modified technique of renal transplantation model in rats.Methods Sprague-Dawley rats were selected as donors ( n=21 ) and Wistar rats as recipients ( n=42 ) .Renal allografts of both sides were harvested from the donors for renal transplantation.After resection of left kidney , end-to-end anastomosis of renal arteries between donor and recipient was performed by the assistance of home-made catheter.And end-to-end anastomosis between recipient's renal vein and donor's inferior vena cava was also performed.The donor's ureter with bladder patch was anastomosed to the recipient's bladder.Finally the right kidney was removed , cefminox (10 mg) was injected intraperitoneally , and then the abdominal cavity was closed.The operation data were recorded , including the operation time , artery and vein anastomosis time , cold and warm ischemia time and so on.Living for 3 days after operation was regarded as a success model.The success rate of modeling was calculated and the cause of death was analyzed.Results The operation time of donor was (32.7 ±5.6) min, and repair time for kidney was (4.2 ±1.1) min.The operation time of recipient was (42.3 ±42.3) min, including (10.1 ±3.2) min of the artery anastomosis time , (13.9 ±2.5) min of vein anastomosis time, (6.3 ±1.4) min of urinary tract reconstruction time.Warm ischemia time was (5.4 ± 1.8) s, and cold ischemia time was (56.2 ±7.3) min.In 42 recipient rats, 40 rats were successful modeling and the success rate was 95.2%.Two rats died.One died of artery anastomosis hemorrhage , and the other died of diffuse peritonitis caused by urine leakage.Conclusions Renal transplantation model in rats with modified vascular end-to-end anastomosis has the characters of simple handling , short operation time and high success rate.

Compared with distal gastrectomy, pylorus-preserving gastrectomy is less invasive which can decrease incidence of dumping syndrome, diarrhea and body weight lost, cholecystitis and gallstone, reflux gastritis and esophagitis and remnant gastric cancer. Based on new Japanese Gastric Cancer Treatment Guideline and new progression in the world, we give a review mainly basic characteristics, indications, operation details and short- and long-time outcomes after pylorus-preserving gastrectomy.
Gastrectomy ; methods ; Gastric Stump ; pathology ; Gastroenterostomy ; Humans ; Organ Sparing Treatments ; Pylorus ; surgery ; Stomach Neoplasms ; surgery

Objective To investigate the correlation between red cell volume distribution width (RDW) and the mortality rate of acute respiratory distress syndrome (ARDS) patients after renal transplantation. Methods Clinical data of 106 ARDS patients undergoing renal transplantation were retrospectively analyzed. According to RDW, all patients were assigned into the normal (≤15.0%, n=68) and increasing RDW groups (>15.0%, n=38). Baseline data and the incidence of adverse events were statistically compared between two groups. Kaplan-Meier survival curve was adopted to compare the 50 d-mortality rate between two groups. Cox's proportional hazards regression model was utilized to identify the risk factors of the mortality of ARDS patients. Results Among 106 patients, the 50 d-mortality rate was calculated as 43.4% (46/106). The sequential organ failure assessment (SOFA) score, serum creatinine, hemoglobin and platelet count significantly differed between two groups (all P<0.05). In the increasing RDW group, the 50 d-mortality rate and the incidence of infectious shock were significantly higher than those in the normal RDW group (both P<0.05). Kaplan-Meier survival curve demonstrated that the 50 d-mortality rate significantly differed between two groups (P<0.01). Cox's proportional hazards regression model univariate analysis revealed that hemoglobin level<100 g/L, serum creatinine>133 μmol/L, platelet count<100×109/L, severe ARDS and RDW>15.0% were the potential risk factors of the 50 d-mortality rate in ARDS patients (all P<0.05). Multivariate analysis demonstrated that severe ARDS [odd ratio (OR)=12.77, 95%confidence interval (CI) 11.63-15.39, P<0.001] and RDW>15.0% (OR=2.01, 95%CI 1.02-3.94, P<0.043) were the independent risk factors of the 50 d-mortality rate in ARDS patients. Conclusions RDW elevation is correlated with the severity of disease and 50 d-mortality rate in ARDS patients following renal transplantation. RDW can serve as a clinical parameter to predict the prognosis of ARDS patients after renal transplantation.

Claudin18.2(CLDN18.2)is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer.It has been identified as a promising target and a potential biomarker to diagnose tumor,evaluate efficacy,and determine patient prognosis.TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2.In this study,we constructed a solid target radionuclide zirconium-89(89Zr)labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines.The[89Zr]Zr-des-ferrioxamine(DFO)-TST001 showed high radiochemical purity(RCP,＞99％)and specific activity(24.15±1.34 GBq/μmol),and was stable in 5％human serum albumin,and phosphate buffer saline(＞85％RCP at 96 h).The EC50 values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058 nM(P＞0.05),respectively.The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors(1.11±0.02 vs.0.49±0.03,P=0.0016)2 days post injection(p.i.).BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i.with[89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups.Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive(+++)for CLDN18.2,while those in the BGC823 group did not express CLDN18.2(-).The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice(2.05±0.16％ID/g)than BGC823 mice(0.69±0.02％ID/g)and blocking group(0.72±0.02％ID/g).A dosimetry estimation study showed that the effective dose of[89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq,which is within the range of acceptable doses for nuclear medicine research.Taken together,these re-sults suggest that Good Manufacturing Practices produced by this immuno-positron emission tomog-raphy probe can detect CLDN18.2-overexpressing tumors.

Objective To optimize a method for extracting traditional Chinese medicine composition with insomnia,and to prepare the insomnia granules for quality control.Methods The optimal extraction process was screened by orthogonal test using high-performance liquid chromatography with geniposide as the evaluation index.The particle size,bulk density,angle of repose,moisture,solubility,hygroscopicity and loading difference of the insomnia granule were evaluated,and the difference between the trial test and the pilot test were analyzed to comprehensively monitor the quality of the insomnia granule.Results The best extraction process was to add 10 times of water and cooked it three times for 1.5 hours each time.The average yield rate of dry extract of the pilot test and trial test was 22.10％,15.52％,and the average yield of powder was 84.96％ and 93.12％,respectively.The conversion rate from the pilot test to the trial test is 76.97％.Both the trial test and the pilot test particles met the quality requirements of the 2015 edition of the pharmacopoeia.Conclusions The preparation method of the insomnia granules is simple and the quality is uniform.The results of the pilot scale showed that the conversion rate is high,the quality is controllable,and the technical feasibility of industrial production is obtained.