10.3969/j.issn.2095-4344.2013.25.025

Objective To investigate the detectable significance of multiparameter flow cytometry (MFC) for the first visiting and minimal residual disease (MRD) in the patients with multiple myeloma .Methods MFC was used to identify the plasma cells by the expression of CD138 or CD38 antigen in 74 patients with multiple myeloma .By combining surface antigens like CD45 ,CD56 , CD19 ,CD20 ,CD117 and the cytoplasm Kappa and Lambda light chain ,the aberrant myeloma cells were differentiated from normal plasma cells .Results In the 44 first visiting cases ,the positive expression of CD138 can be detected in all cases ,while the expres‐sion of CD19 was negative and 42 cases (95% ) were negative or weak positive expression for CD45 .The detection rates of CD38 , CD56 ,CD20 and CD117 were 98% ,93% ,45% and 41% ,respectively .The cytoplasm Kappa and Lambda light chains were showed the limited expression .Of the patients with MM ,14 cases were used for evaluating the change of immunophenotype at first visiting and during the treatment process ,among them ,11 cases(79% ) appeared the changes in at least one of aberrant phenotypes .4 cases (29% ) had the significant enhancement of antigen marker fluorescence intensities after chemotherapy and 7 cases (50% ) had sig‐nificant decrease of antigen marker fluorescence intensities after chemotherapy .CD45 ,CD19 and cytoplasm immunoglobulin light chains were the most stable marker ,no obvious antigen marker changes were found during the treatment ,while there was a signifi‐cant antigen density change in 2 cases of CD38 (14% ) ,7 cases of CD56 (50% ) ,4 cases of CD20 (29% ) and 2 cases of CD117 (14% ) .Of the 30 cases for evaluating MRD immunophenotype ,the abnormal myeloma cells were detected in 25 cases .In 5 cases ,no expression of limited Kappa and Lambda light chains was found and the ratio of Kappa and Lambda was 0 .5 - 2 ,which were identi‐fied as negative for MRD .Conclusion The multiparameter flow cytometry has important significance in evaluating the diagnosis , therapeutical effect and prognosis .The detection by adopting cytoplasm immunoglobulin light chains can improve the accuracy in MRD detection .

Objective To study the frequency of the derivative chromosome 9 [der(9)] deletion among patients with chronic myeloid leukemia (CML), and explore the application value of local-specific inspector (LSI) 9q34 probe in detect the der (9) deletion. Methods Cytogenetic analysis was performed by 24 h unstimulated culture, GTG banding and karyotype. Dual colour/dual fusion or extra signal(ES) DNA probe was used to perform interphase-FISH for the detection of bcr-abl fusion gene in 52 patients with CML. LSI 9q34 DNA probe was used to detect der(9) deletion. Results FISH results showed bcr-abl fusion gene existed in all 52 patients with CML. der(9) deletion was detected in 12 patients with ES/DCDF probe, but only in 11 patients with LSI 9q34 probe. Conclusion It's more efficient in detection of der(9) deletion with LSI 9q34 probe. Deletion of der(9) might be a poor prognostic factor for CML patients, and LSI 9q34 probe should be used in all the CML patients with positive bcr-abl fusion gene.

Objective To study the change of glycogen phosphorylase isoenzyme BB(GPBB)in neonates with asphyxia complicated with myocardial injury and its correlation with various perinatal factors. Methods Sixty-four neonates with asphyxia(including 39 mild asphyxia and 25 severe asphyxiated neonates,30 neonates with and 34 without myocardial injures)were enrolled and 25 healthy neonates were studied as control. The plasma levels of GPBB were measured by enzyme-linked immunosorbent assay(ELISA). Myocardial enzymes,cardiac troponin I,electrocardiogram,chest X-ray were performed simultaneously. Results The plasma GPBB levels were significantly higher in neonates with myocardial injury(13.84,7.57 ng/ml)than those without myocardial injury(4.97,3.24 ng/ml)and control group(4.95,1.99 ng/ml)( P ＜ 0.01). The sensitivities of GPBB,cTnI and CK-MB in diagnosing myocardial injury were 90%,66.7% and 83.3%,respectively. The sensitivity of GPBB was significantly higher than that of cTnI(χ~2 ＝ 4.812,P ＜ 0.05),with no statistical difference between GPBB and CK-MB(χ~2 ＝ 0.577,P ＞ 0.05). The specificities of GPBB,cTnI and CK-MB in diagnosing myocardial injury were 88.2%,91.2% and 67.6%,respectively. The specificity of GPBB was significantly higher than that of CK-MB(χ~2 ＝ 4.191,P ＜ 0.05),with no statistical difference between GPBB and cTnI(χ~2 ＝ 0.159,P ＞ 0.05). Plasma GPBB levels were significantly higher in neonates with severe asphyxia(14.67,6.09 ng/ml)than those with mild asphyxia(5.61,3.56 ng/ml)and control group(P ＜ 0.01). GPBB levels were higher in mildly asphyxiated neonates than those in control neonates,but with no statistical significance(P ＞ 0.05). Spearman rank correlation analysis showed that plasma GPBB levels positively correlated with the cloudiness of the amniotic fluid(r ＝ 0.500,P ＜ 0.001)and negatively correlated with Apgar score(r ＝ -0.520,P ＜ 0.001). Conclusions This study indicated that GPBB can be used as an ideal biomarker of myocardial ischemia injury in neonates with asphyxia. The determination of GPBB in early neonatal period is useful in detecting and assessing the severity of myocardial injury.

Objective To investigate the effects of transient plateau factor on acute lung injury induced by phosgene poisoning in rabbits.Methods Forty New Zealand white rabbits of both sexes,aged 2.0-2.5 kg,were randomly divided into 4 groups (n =10 each) using a random number table:control group (group C),plateau factor group (group H),phosgene poisoning group (group P),and phosgene poisoning and plateau factor group (group HP).In group H,the rabbits were exposed to a simulated altitude of 33000 m for 2 h.In group P,the rabbits were exposed to phosgene for 3 min only.In group HP,the rabbits were exposed to phosgene for 33 min and then to a simulated altitude of 3000 m for 2 h.Respiratory rate (RR) was recorded and blood samples were taken before exposure to phosgene (T1),after exposure to phosgene (T2),and at 0,1 and 6 h after onset of exposure to a simulated altitude of 33000 m (T3-5) for determination of PaO2 and oxygenation index (OI) was calculated.The chests were opened at T5 and lungs removed for determination of lung water content (LC) and for microscopic examination.Lung coefficient (LC) was calculated.Results Compared with C group,RR was significantly increased at T3 in group H (P ＜ 0.05),and RR was increased and OI was decreased at T2-5 in P and HP groups (P ＜ 0.05 or 0.01).Compared with P group,RR was increased and OI was decreased at T3-5 in HP group (P ＜ 0.05 or 0.01).LW and LC were significantly higher in P and HP groups than in group C,and in HP group than in group P (P ＜ 0.05 or 0.01).The microscopic examination showed that pathological changes were observed in P and HP groups,however,the changes were severer in HP group.Conclusion Transient plateau factor can obviously aggravate the degree of acute lung injury induced by phosgene poisoning in rabbits.

Objective: To investigate the proper methods of acupuncture and moxibustion for controlling body weight and their objects. Methods: 274 outpatients were allocated to 3observation groups: obesity, overweight and normal, using standard weight as an index. The obesity group was divided, at a ratio of 1: 1: 3, into 3 groups: fixed prescription, local treatment and treatment based on syndrome differentiation. The syndromes were classified into five types in the syndrome differentiation group. Different protocols of acupuncture and moxibustion plus auricular-plaster therapy were drawn up for treatment. Body mass index (BMI), body fat (F%),body circumferences and subcutaneous fat were evaluated in the patients before and after treatments. Results: There was a difference in the controlling effect of acupuncture on body weight among different populations. The curative effect was better in the obesity and overweight groups than in the normal group; the effective rate was 72.2%, 72.5% and 44.0%, respectively.There was also a difference in curative effect among obesity patients with different types of syndromes. The curative effects on retention of damp-heat in the stomach, stagnation of liver qi and dampness retention due to splenic hypofunction were equal but all better than those on deficiency of splenonephric yang and intemal heat due to yin deficiency. There was also a difference in curative effect among different protocols. The curative effects in the syndrome differentiation-based treatment and fixed prescription groups were equal (77.8%) but all better than that in the local treatment group (53.3%). Conclusion: Acupuncture and moxibustion is an effective way to treat obesity and overweight. Treatment based on syndrome differentiation and fixed prescription on the basis of syndrome differentiation are effective methods.

Objective To investigate the role of C-Jun N-terminal kinase (JNK) in epithelial mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells(RPMCs). Methods RPMCs were harvested from the peritoneum of male Sprague-Dawley rats, then cultured in DMEM/F12 medium with 15% (V/V) FBS. After stimulation with TGF-β1, the expression of a-smooth muscle actin (α-SMA), E-cadherin and collagen I were detected in RPMCs. In some groups, the ceils were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGF-β1. The protein expression of phosphorylated JNK was detected by Western blotting. The mRNA and protein expression ofα-SMA, E-cadherin and collagen I were examined with RT-PCR and Western blotting, respectively.The intracellular distribution and expression of α-SMA was determined by indirect immunofluorescence. Results TGF-β1 could significantly increase the expression of α-SMA and collagen I, and decrease the expression of E-cadherin in RPMCs. TGF-α1 could stimulate the expression of phosphorylated JNK at 5 minutes with the peak at 10 minutes (P<0.01). The addition of SP600125 effectively inhibited TGF-β1-induced high expression of α-SMA and collagen I (P<0.05), and prevented TGF-β1-induced down-regulation of E-cadherin expression in RPMCs (P<0.05). The indirect immunofluorescence showed that the expression of intracellular α-SMA in RPMCs stimulated by TGF-β1 for 48 h increased significantly, which could be inhibited by SP600125. Conclusions JNK regulates epithelial mesenchymal transition induced by TGF-β1 in rat peritoneal mesothelial cells. JNK inhibitor may be used as a novel therapeutic agent for peritoneal fibrosis.

Objective To investigate the micminnammatory state and its relationship with atherosclerosis and cardiac function in patients on continuous ambulatory peritoneal dialysis(CAPD). Methods Sixty-seven CAPD patients,27 non-dialytic stage 5 chronic kidney disease(CKD 5)patients and 27 gender and age matched healthy controls wore enrolled in this cross-sectional study.Clinical data and biochemical parameters were collected.Serum interleukin-6(IL-6)and IL-10 levels were measured by enzyme-linked immunosorbent assay(ELISA).High-sensitivity C-reactive protein(hs-CRP)was measured by immunoturbidimetry.Prevalence of atherosclerosis was detected by carotid uhrasonography while cardiac function was detected by echocardiography. Results Serum levels of inflammatory biomarkers were elevated significantly in CAPI)and CKD 5 patients as compared with healthy controls[IL-6(ng/L):2.400,1.515 vs 0.698;IL-10(ng/L):1.988,1.958 vs 0.277;hs-CRP(mg/L):1.090,1.345 vs 0].Left ventricular mass index(LVMI),myocardial performance index(Tei index 0.75±0.31,0.66±O.27 vs 0.52±0.23)in CAPD and CKD 5 patients increased significantly.The prevalence of carotid artery atherosclerosis and left ventricular hypertrophy(LVH)in CAPD and CKD 5 patients was significantly higher than that in healthy controls.No significant difference of the ultrasonic parameters was found between CAPD and CKD 5 patients.In CAPD patients,IL-6 was positively correlated with Tei index,whereas IL-10 was negatively correlated with INMI and was positively correlaled with ejection fraction(EF).In a multiple regression model,IL-6,self-rating depressive scale(SDS)score and pulse pressure were independent predictors of carotid artery atherosclerosis.Similarly,IL-6 and primary hypertension were independent correlates of Tei index in CAPD patients. Conclusions Microinflammatory state exists in either non-dialytic CKD 5 patients or CAPD patients and it is associated with atherosclerosis and cardiac distunetion.IL-6 is an independent risk factor of atherosclerosis and increased Tei index in CAPD patients.

Objective To explore the status of histone acetylation modification and their regulatory effect to hMSH2 gene and hMLH1 gene expression in acute leukemia. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of hMSH2 and hMLH1 mRNA, and Western blot was used to measure the expression of histone H3, H4, HDACi, hMSH2 and hMLH1 protein in mononuclear cells of 56 acute leukemia patients and 30 healthy volunteers. The mononuclear cells of 30 acute leukemia patients were treated with histone deacetylase inhibitors trichostatin A (TSA), and measured the expression difference of histone H3, H4, HDAC1, hMSH2 and hMLH1 in the mononuclear cells treated with TSA. Results The protein expression levels of hMSH2, hMLH1, histone H3 and histone H4 in those mononuclear cells of acute leukemia patients were 0.4610±0.1211, 0.4013±0.1143, 0.4103±0.1241 and 0.4251±0.1081, respectively, which were significantly decreased comparing with those of healthy volunteers (0.9461±0. 1841, 0.996±0.2021, 0.8971±0. 1194 and 0.9513±0.1953) (t = 3.341, 3.935, 2.843 and 3.575,respectinely, P ＜0.05). The protein expression levels of HDAC1 (0.8841±0.2018) of acute leukemia patients was significantly increased comparing with those of healthy volunteers (0.5142±0.1340) (t= 2.634, P ＜0.05).After treatment with TSA for 48 hours, the protein expression of hMSH2 was increased nearly 1.5-fold, hMLH1 about 1.6-fold, H3 about 2.9-fold and H4 about 3.4-fold comparing with the negative control groups (P ＜0.05),while the protein expression of HDAC1 were decreased comparing with the negative control groups by 40 %.Conclusion There was an low expression phenomenon of histone acetylation in acute leukemia, and histone acetylation played an important role in regulation of the mismatch repair gene expression in acute leukemia.

Objective To investigate the safety and feasibility of reoperatively laparoscopic technique in treatment for locally recurrent rectal cancer.Methods The study enrolled 17 patients with locally recurrent rectal cancer between February 2004 and September 2009 from Shanghai Minimally Invasive Surgery Center.The patients were divided into two groups according to their pelvic recurrence types:central recurrence group (n =14) and anterior recurrence group (n =3).Demographic,surgical data and survival outcomes between two groups were compared.Results The outcomes of demographic data between two groups were not different(P＞ 0.05 ).Compared with central recurrence group,anterior recurrence group had longer operating time (P =0.028).However,the differences of operative blood loss,complications,postoperative rehabilitative outcomes and ratio of R0 resction between groups were not significant ( P ＞ 0.05 ).The overall 5- year survival rate of all the patients was 36％.And the median survival time was 42 months without significant difference between two groups (x2 =1.641,P =0.200).Conclusions Reoperatively laparoscopic technique in treatment for locally recurrent rectal cancer is safe and feasible.Selected patients,specialist operation and higer ratio of R0 resection are the key factors conducive to better short-and long-term outcomes.