20 adult healthy rats were used for microelectropheretic delivery of horseradish peroxidase (HRP). The diameter of the tip of the glass electrode was around 20～50?. It was inserted into nucleus raphe magnus, 20% HRP was delivered microelectrophoretically(5 ?A for 30 min; 10 ?A for 10 min). In eight rats the microelectrode was withdrawn immediately after the delivery and in the rest it was removed 10～20 minutes after delivery. Afterward the animals were allowed to survive for 40 hrs before decapitation, except 2 for 20 and 2 for 60 hrs. The brains were dissected out for cryostat sections and DAB reaction for peroxidase.The tip of the electrode was located in the nucleus raphe magnus, at the level of nucleus facialis. The diameter of the brain tissues occupied by HRP at the points of microelectophoretic delivery was 0.2～0.4mm and the maximum reached 0.6～0.7mm. The exogenous HRP granules were not visible in the cases with electrodes withdrawn immeditely after delivery and in the survivors of 60 hrs. In most cases the electrodes were left inside for more than 10 minutes after the delivery. The exogenous HRP granules were large, coarse, brown, steroscopic and distributed evenly in the perikaryon of some neurons in several levels of the brain. Those in the axon and dentrite were distributed just like strings of pearls. There were much more neurons with exogenous HRP positive granules dispersed in the reticular formation of the medula oblongata, most of them were polydendritic. Small and fusiformed HRP-positive cells are observed occasionally in the gray substance of subventriculum, reticular formation pontis and ventrolateral side of locus coeruleus. Weaker reactions were found in a few cells in the periaquaductous gray substance of the midbrain, nucleus raphe centro-superior, substantia nigra pars compacta and in the area around the nucleus supraopticus. Besides no HRP-positive cells were discovered in thalamus and subcortex structure.There were two types of cells with HRP positive granules around some blood vessels in reticular formation and subventricular gray substance: One was a small protoplasmic astrocyte with broad cytoplasma, small nucleus, and thick, short and irregular processes and another kind was polydendritic neurons filling with brown positive granules in perikarya and cytoprocesses, in the latter, the HRP positive granules were arranged as strips of pearls.

Objective To analyze the genetic diversity of Psammosilene tunicoides,an endangered and endemic medicinal plant,in southwest China.Methods The genetic diversity of seven representational populations of P.tunicoides including 137 individuals had been investigated by amplified fragment length polymorphisms(AFLP)maker technique.Results The genetic diversity had been revealed as follow:the Nei's genetic diversity index(He),Shannon's information index(I),percentage of polymorphic loci(PPB)were 0.243 4?0.179 1,0.373 5?0.248 5,and 82.30%,respectively at the species level;and 0.091 8?0.161 0,0.140 2?0.236 2,and 30.48%,respectively at population level.The genetic differentiation index(Gst)was 0.624 6 and genetic differentiation coefficient by Shannon's diversity(Ist)was 0.624 6.The result of dendrogram of seven populations indicated that Lijiang and Gejiu of Yunnan populations shared the maximum genetic identity,though they distributed in a relatively great geographical distance;Kunming population of Yunnan had the greater genetic distance from other populations.Nine characteristic fingerprint bands that can distinguish the different populations had been acquired.Conclusion The genetic diversity of P.tunicoides is relatively higher at the species levels,while lower within population levels,and a significant degree of genetic differentiation occurrs among the populations.There is little relativity between the relationship of populations and geographical distance.The combination of characteristic fingerprint bands between intraspecies and populations provide important basis data for germplasm resources diagnostics and plant breeding by AFLP maker technique.

Objective:To explore the clinical characteristics, diagnosis and treatment of acute leukemia patients during pregnancy.Methods:The clinical data of 16 cases with acute leukemia during pregnancy from January 2009 to December 2018 in the First Affiliated Hospital of USTC were retrospectively analyzed. The diagnosis and treatment regimens, pregnancy outcome, the early fetus and survival status of patients were also analyzed.Results:All 16 leukemia cases were confirmedly diagnosed and classified by bone marrow puncture, including 13 cases of acute myeloid leukemia (5 cases of non-acute promyelocytic leukemia and 8 cases of acute promyelocytic leukemia) and 3 cases of acute lymphoblastic leukemia. At the time of confirmed diagnosis, 6 patients were in first trimester, 6 cases in second trimester and 4 cases in late trimester. As for pregnancy outcome, 1 patient had natural birthing, 5 patients underwent cesarean operation, 9 patients underwent artificial abortion and 1 patient had spontaneous abortion. Chemotherapy was performed in 15 patients during pregnancy, 11 patients received chemotherapy for treatment of primary disease after pregnancy, 3 patients died during the treatment. During the follow-up of 13 cases, 8 patients survived and 5 patients lost follow-up.Conclusions:Early diagnosis of acute leukemia during pregnancy is very important. Bone marrow puncture should be performed timely to make clear diagnosis when blood routine result is abnormal during antenatal care. Multidisciplinary consultation should be initiated in time, and the best treatment plan should be worked out to guard against serious complications during pregnancy.

Objective To investigate the efficacy and safety of posaconazole in the prophylaxis or salvage treatment of invasive fungal disease (IFD) in patients with hematological diseases.Methods 25 patients with hematological diseases receiving posaconazole treatment from Feb 2014 to Feb 2015 were analyzed retrospectively.The patients' average age was 32.6 years old (16-64 years old).18 patients received posaconazole for IFD prophylaxis, and 7 patients for IFD salvage treatment.Results 18 patients receiving posaconazole for IFD prophylaxis had no clinical manifestation of IFD (that is no case of breakthrough fungal infection) during treatment or in the 12 weeks after treatment, and the average prophylaxis period was 21 d (14-35 d).Among 7 patients receiving posaconazole for IFD salvage treatment, 6 patients were effective, including 4 cases cured and 2 cases effective.There were no obviously side effects of posaconazole in these patients.Conclusion Posaconazole has good clinical response for IFD in the hematologic diseases patients whether in prophylaxis or in salvage treatment.

Objective To set up a real-time quantitative PCR approach for detection and quantification for bcr-abl transcripts in CML patients,and detect minimal residual disease (MRD) in CML by real-time quantitative PCR (RQ-PCR)and evaluate the significance of MRD detection.Methods The ber-abl.fusion gene expression in 80 patients with CML was analyzed by RQ-PCR. The patients were divided into three groups according to the different treatment, allogeneic hematopoietic stem cell transplantation group,imatinib group and hydroxyurea group. The change of bcr-abl fusion gene was monitored in CML patients before and after treatment.Results The average of RQ-PCR detection on newly diagnosed patients with CML in chronic phase was 6847.67 copies / 104 cells,the accelerated phase was 306 176.08 copies / 104 cells,and the average results were 944.33, 2.37, 0.29, 0 copies / 104 cells after allogeneic hematopoietic stem cell transplantation one month,6 months,12 months or 24 months respectively.The average of RQ-PCR detection after use imatinib mesylate 3 months was 3720.23 copies / 104 cells and not be detected after one year. The average was 7290.11 and 3143.24 copies / 104 cells after hydroxyurea treatment 0 and 9 months respectively.The difference in first two groups was not significant (t=1.74,P=0.17), but the difference between the third group and the first two groups was significant (t=3.74,P=0.01.t=2.97,P=0.02). The upregulation of bcr-abl transcript levels could be detected when disease progression. The transcripts level in accelerated phase was significantly higher than that in chronic phase. Conclusion RQ-PCR can be used to detect the MRD,monitor the treatment outcome,predict disease recurrence and give early intervention.

ObjectiveTo explore the genetic abnormalities of multiple myeloma (MM)patients by fluorescence in situ hybridization (FISH).MethodsWith the application of FISH,sequence specific DNA probes (IGH,DI3S319/p53 and 1q21/RB1) were applied to detect 14q32 rearrangement,del(13q14),del (17p13)and gain of lq21.Forty-four MM patients were enrolled in this study.ResultsThirty-two cases (72.7 ％) detected by FISH had genetic abnormality in 44 cases,lq21 amplification was observed in 11 cases (25.0 ％),while RB1 deletion in 17 cases (38.6 ％),D13S319 deletion in 16 cases (36.4 ％),p53 deletion in 6 cases(13.6 ％)and 14q32 translocation in 19 cases(43.2 ％).The patients with one abnormality was detected in 10 cases(22.7 ％),two abnormalities in 11 cases(25.0 ％),three abnormalities in 8 cases (18.2 ％),4 abnormalities in 3 cases(6.8 ％).28 were found to undergo split-phase by conventional cytogenetic in 44 patients.The patients with genetic abnormalities detected by conventional G-banding was 2 cases (7.14 ％),the difference with that in FISH was significant (P ＜0.05).Genetic abnormalities compared with clinical parameters showed that β2-MG in IGH gene abnormal patients were significantly higher than those without such abnormalities (P ＜0.05).Patients with bone marrow plasma cells of lq21 amplification were higher than those with normal karotypes(P ＜0.05),CRE was significantly higher among lq21 amplification and p53 deletion patients (P ＜0.05),CRP was significantly higher among p53 deletion patients (P ＜0.05).No significant difference was oberserved in relationship of the chromosome aberration and age,the chromosome aberration and stage.ConclusionThe most common genetic abnormalities in MM is IGH rearrangement and absence of RB1 and D13S319,followed by lq21 amplification,the least is p53 deletion.FISH is a rapid and sensitive technique to refine chromosome aberrations in MM.The specific detection for genomic features of MM is proved to be correlative with its clinicopathologic characteristics and the prognosis.

The objective of the study is to find out the synergistic effect on apoptosis resulting from the combination of chemotherapeutic drugs and some cytokines. Dexamethasone (DEX), etoposide (VP16), arsenic trioxide (As(2)O(3)) and alltrans-retinoic acid (ATRA) were added to the murine T lymphoma cell line RMA as well as to the cells preincubated with IL-2, IL-6 or GM-CSF, respectively. The effect on apoptosis was observed. All four chemotherapeutic drugs inhibited the cell proliferation. DEX, VP16 or As(2)O(3), except ATRA, singly induced apoptosis of RMA cells. The DEX concentration of inducing apoptosis was reduced when it was added together with ATRA. IL-2, IL-6 and GM-CSF did not induce apoptosis when the cytokines were added to RMA separately, however, apoptosis could be induced by combination of IL-2 and IL-6. The cytokines facilitated the apoptotic action of chemotherapeutic drugs, the drug concentration for inducing apoptosis decreased and the time period of starting apoptosis shortened. The results demonstrated that there was synergistic effect of chemotherapeutic drugs and some cytokines for induction of apoptosis. It raised an experimental basis for combination of chemotherapeutic drugs and some cytokines, especially IL-2, in the treatment of malignant lymphoma.

Objective To investigate the effects of the erythropoietin (EPO) on ischemia reperfusion injury (IRI) in rats with nephron-sparing surgery (NSS). Methods Fifty-four Sprague Dawley rats were divided into 3 groups randomly after right kidney nephrectomy: Sham group, NSS group (PBS+NSS) and EPO group (EPO+NSS). During NSS, renal artery was clamped for 40 min to induce IRI. Sham group just adopted exposure renal artery without vascular clamped. Rats in NSS group were injected intraperitoneally with PBS for 3 days before NSS. Rats in EPO group were injected intraperitoneally with EPO for 3 days before NSS. After 12 h, 24 h, 72 h, blood sample and renal tissues were collected. The serum creatinine (Scr) and urea nitrogen (BUN) were evaluated. The pathology injury was evaluated by HE staining. The CD24/CD133 double-positived renal progenitor cells (RPCs) were tested by flow cytometry. The CD133 and PCNA protein were quantified by immunohistochemical staining. The expressions of Wnt7b and β-catenin protein were detected by Western blotting. Results Rats in NSS group had more elevated Scr, BUN and pathology injury scores 12 h, 24 h and 72 h after operation than those in Sham group (all P<0.05). Compared with those in the NSS group, the Scr and BUN in the EPO group were significantly lower 24 h after the surgery (all P<0.05), and the pathology injury score also decreased (P<0.05). The proportion of RPCs, expressions of CD133 and PCNA, and expressions of Wnt7b and β-catenin protein were significantly higher after 24 h of the surgery in NSS group than those in the Sham group (all P<0.05). While compared with those in the NSS group, the proportion of RPCs and expressions of CD133, PCNA, Wnt7b and β-catenin increased at the EPO group (all P<0.05). Conclusions EPO can reduce the IRI after NSS, and its mechanism may be related to the mobilization of the RPCs by the Wnt7b/β-catenin signal pathway.

Objective: To compare the efficacy of unrelated cord blood transplantation (UCBT) and HLA-identical sibling peripheral blood stem cell transplantation (PBSCT) for the treatment of adult hematological malignancies. Methods: From April 2011 to December 2015, a total of 81 patients receiving single-unit UCBT and 57 patients receiving HLA-identical sibling PBSCT were enrolled in this study. All of the patients received myelablative conditioning. Cyclosporine combined with mycophenolate mofetil was adopted for GVHD prophylaxis. Results: The cumulative incidence of neutropil engraftment at day-42 was 95.0% and 100% in UCBT and sibling PBSCT groups, respectively (P=0.863) . Platelet engraftment at day 100 was 87.3% (95%CI 76.8%-93.1%) in UCBT group, which was significantly lower than that of sibling PBSCT group[98.2% (95%CI 87.3%-99.7%) ] (P=0.005) . There were no significant differences in terms of Ⅱ-Ⅳ acute GVHD or Ⅲ-Ⅳ acute GVHD in two groups (P=0.142, 0.521) . The 3-year chronic GVHD and extensive chronic GVHD were 14.9% (95%CI 5.2%-23.5%) and 11.2% (95%CI 2.9%-18.7%) , respectively in UCBT group, which was significantly lower than that of sibling PBSCT group[35.2% (95%CI 19.4%-47.8%) , 31.4% (95%CI 16.2%-43.9%) ] (P=0.008, 0.009) . The 3-year transplant-related mortality (TRM) was similar between two groups (30.1% vs 23.2%, P=0.464) . The relapse rate at 3-year in UCBT group[12.9% (95%CI 6.6%-21.5%) ]was significantly lower than that in sibling PBSCT group[24.3% (95%CI 13.5%-36.8%) ] (P=0.039) . There were no significant differences in terms of overall survival (OS) and disease-free survival (DFS) between two groups (58.6% vs 54.8%, P=0.634; 57.0% vs 52.4%, P=0.563) . But GVHD-free and relapse-free survival (GRFS) in UCBT group [55.7% (95%CI 44.1%-65.8%) ]was significantly higher than that of sibling PBSCT group[42.9% (95%CI 29.8%-55.3%) ] (P=0.047) . Conclusions For adult hematological malignancies, the incidences of acute GVHD and TRM were similar between UCBT and sibling PBSCT recipients, and the incidences of chronic GVHD and relapse were lower in UCBT recipients. UCBT recipients had higher GRFS rate although OS and DFS were similar between two groups, which may reflect the real recovery and better quality of life following UCBT.

Objective To study the curative efficacy and safety of single-unit umbilical cord blood transplantation (sUCBT) for malignant hematologic diseases,which is provided by China's public cord blood bank.Methods We retrospectively analyzed 409 cases of malignant hematologic diseases who accepted myeloablative single-unit unrelated donor UCBT without ATG at our center between May 2008 and December 2016.A comparative analysis was made on the total nuclear cells (TNC) of the umbilical cord blood before freezing and after thawing,the cells of CD34+,the recovery rate of cells and the clinical effect of UCBT.Result 409 units of umbilical cord blood used in UCBT respectively came from eight China's public cord blood banks.The average TNC of 409 units of umbilical cord blood before freezing and after the tubular recovery were respectively 18.5 × 108 and 16.34 × 108 (p =0.000).The average recovery rate of the tubular recovery was 88.5％,and there was significant difference among cord blood banks (P =0.000).The average TNC of umbilical cord blood before freezing and transfusion were respectively 18.5 × 108 and 15.86 × 108 (p =0.000).The average recovery rate of umbilical cord blood transfusion was 85.9％,with the difference being significant among cord blood banks (P =0.000).The average number of CD34+ cells before freezing and after the tubular recovery was 11.18 × 106and 8.68 × 106 (p =0.000).The average recovery rate of CD34+ cells after the tubular recovery was 80.75 ％,with the difference being significant among the cord blood banks (P =0.000).At 42nd day after UCBT,the cumulative incidence of neutrophil engraftment was 95.4％,and the median time of the engraftment was 17 days (11-38 days).The cumulative incidence of platelet engraftment at 120th day was 84.6％,and the median time of the engraftment was 36 days (14-93 days).The cumulative incidence of erythrocyte engraftment at 60th day was 92％,and the median time of engraftment was 22 days (9d-60 days).After the umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of engraftment.The P values for cumulative incidence of neutrophil,platelet and erythrocyte engraftment were respectively 0.004,0.01 and 0.000 2,with the differences being statistically significant.At 100th day after UCBT,the cumulative incidence of Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees of acute graft-versus-host disease (aGVHD) was respectively 28.63％ and 15.7％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of aGVHD.There was no significant difference between Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees (P =0.809 and 0.68 respectively).At 3rd year after UCBT,the cumulative incidence of relapse was 15.89％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the cumulative incidence of relapse (P =0.898).At 3rd year after UCBT,the overall survival (OS) rate and disease free survival (DFS) rate were respectively 66.7％ and 59％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in OS and DFS.There was no significant difference in OS and DFS (P =0.566 and 0.703 respectively).At 3rd year after sUCBT,the rate of graft-versus-host diseases/relapse-free survival (GRFS) was 54.3％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the rate of GRFS (P =0.449).Conclusion The umbilical cord blood provided by China's public cord blood bank was used in UCBT.It has a high safety and good efficacy in treating malignant hematologic diseases.But it needs to set up the standardized and normalized quality-control system of umbilical cord blood for China's public cord blood bank.