Objective Retrospective investigate the significant application of bone marrow (BM)morphology in exploring the causes of anemia,and observed the changes of anemia disease spectrum in the past 10 years.Methods BM smears of 1 528 anemia patients from Shanghai Jiao Tong University Affiliated Shanghai First People's Hospital were stained with Wright's staining and cvtochemical staining and observed with microscope.Combined with relevant clinical data,we analyzed the changes of disease types from 1998 to 2002 and from 2008 to 2012.Results 1 139 cases (74.54％) were diagnosed with hematopoietic and lymphatic system diseases.Iron-deficiency anemia,megaloblastic anemia and leukemia were the three main causes of anemia.The BM morphology of 389 patients displayed infection anemia or descriptive diagnosis.Anemia disease spectrum changed a lot,iron-deficiency anemia decreased from 30.34％ (139/458) to 18.69％ (200/1 070),leukemia increased from 13.31％ (61/458) to 21.77％ (233/1 070),descriptive diagnosis of BM increased from 15.72％ (72/458) to 21.86％ (234/1 070).Conclusion BM examination is critical for finding the cause of anemia,which contrihutes to effective treatment.

OBJECTIVE: To explore the effect of different pressure on microcirculation of scar proliferation. METHODS: A total of 60 patients with scar proliferation after burning were collected, including 49 males and 11 females, with averagely aged 37 years. The patients were divided into low pressure (0.67-1.33 kPa), normal pressure (1.33-3.33 kPa) and high pressure (3.33-6.67 kPa) groups by envelope method. Pressure tension bandage was used at 1 week after wound healing and gradually increased, reached expected pressure at days 5-7 and sustained for 6 months. Then the changes of blood perfusion at the scar tissue were measured by laser Doppler perfusion imaging (LDPI). RESULTS: According to the intended processing analysis, all the 60 patients enter the result analysis. There were no significant differences of the scar tissue perfusion of the 3 groups before the therapy. After the therapy, the perfusion was decreased in the low pressure group, but there are no statistical changes compared to before the therapy (P > 0.05), while the perfusion was decreased in the normal pressure and high pressure groups (P < 0.05), which was lower in the high pressure group than the normal pressure group (P < 0.05). When the pressure increased to 3.33-6.67 kPa, the patients feel much more uncomfortable and the swelled signs appear, and 2 patients quit the experiment. CONCLUSIONS: The scar tissue perfusion is decreased obviously when adding the pressure to 3.33 kPa by using tension bandage. Using this method, the pressure should gradually increase to the maximum if patients can bear.

Adenoma ; metabolism ; pathology ; surgery ; Humans ; Keratin-7 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Kidney Transplantation ; Living Donors ; Male ; Mucin-1 ; metabolism ; Young Adult

AIM:To observe the effect of botulinum neurotoxin type A heavy chain ( BoNT/A HC) on the pat-tern of spinal protein expression by intrathecal injection after spinal cord injury in rats , and to explore the role of BoNT/A HC intervention in spinal protein expression and some of its mechanisms in nerve regeneration after injury .METHODS:The model of unilateral lumbar spinal cord injury was established .The effects of BoNT/A HC intervention at different doses (2 μg, 4 μg, 6 μg and 8 μg) on the general pattern of protein expression in the spinal cord tissues at the injury site and the cranial part adjacent to the injury site was measured and evaluated by SDS-PAGE and Coomassie brilliant blue staining first, and then by two-dimensional SDS-PAGE.RESULTS:The histological structure of the ipsilateral side of lumbar spi-nal cord showed obvious destruction and degradation , mainly affecting both gray and white matter of the left side of the cord.The result of SDS-PAGE with Coomassie brilliant blue staining from injured spinal cord tissue displayed that the ex-pression of some proteins after one-time BoNT/A HC treatment appeared obviously different from that without BoNT /A HC treatment.Moreover, the pattern of the protein expression affected by BoNT/A HC was similar to that of the normal spinal cord.The more detail information from two-dimensional SDS-PAGE indicated that more than 10 proteins with different mo-lecular weight and isoelectronic points were differentially expressed at day 2 and day 20 after local injection of 6μg BoNT/A HC.This altered expression actually appeared a tendency toward the pattern shown in normal group .CONCLUSION:The immediate application of BoNT/A HC at the injury site after unilateral lumbar spinal cord injury is able to affect the pattern of local protein expression .The altered protein expression by injury could be reversed back to normal or approxi -mately normal by local BoNT/A HC administration.

Objective To investigate the effect of small direct‐current electrical stimulation on migration and invasion related MMPs/TIMPs expression of trophoblast cells .Methods The trophoblast cells were exposed to the direct current electrical field at 150 mV/mm for 5 and 10 hours .Cell images were recorded with continuous photographing and analyzed by image analyzer .The ex‐pression levels of MMP2 ,MMP9 ,TIMP1 and TIMP2 were measured using quantitative RT‐PCR and Western blot .Results In non‐electrical field culture trophoblast cells migrated slowly with random directions .Trophoblast cells cultured in media containing 10% calf serum with the application of 150 mV/mm direct current electrical stimulation ,showed marked cathodal migration (P<0 .01) ,the cell body stretched ,perpendicular to the direction of the electric field .Compared with the non‐electrical field stimulation controls ,trophoblasts under the electrical field stimulation had the increased MMP2 mRNA and protein expression (P< 0 .05) , while MMP9 ,TIMP1 and TIMP2 had no obvious changes of mRNA or protein expressions .Conclusion Physiological direct‐cur‐rent electrical fields might induce directed migration and perpendicular orientation of trophoblast cells .The enhanced MMP2 expres‐sion may play an important role in the migration and invasive activity of trophoblast cells in small electrical field .

Objective To compare the effect of intravenous resuscitation with sodium pyruvate (Pyr) Ringer solution against lactated Ringer solution on hemodynamic and organ functions during shock stage in dogs with burn.Methods 28 Beagle dogs were subjected to 50％ total body surface area (TBSA) burn,and they were divided into three groups:burn injury without fluid resuscitation (N R,n =8),Ringer lactate solution(RL,n =10),and Pyr Ringer solution (RP,n =10).They were given intravenous fluid resuscitation according to Parkland formula 30 minutes after burn.The hemodynamics,organ functions and mortality were observed in conscious state before burn injury,and 2,6,8,12,24 hours after burn injury.Results Within 24 hours after burn,all the dogs in NR group died,and those in RL and RP groups were all alive.At 2 hours after burn,the mean arterial pressure (MAP),cardiac index (CI),dp/dt max of left ventricular contractility were significantly reduced in NR,RL and RP groups compared with those before injury [MAP(mmHg,1 mmHg =0.133 kPa):45.33 ± 7.78 vs.141.67 ± 5.98,91.33 ± 10.25 vs.142.33 ± 6.16,98.67 ± 9.54 vs.142.83 ±5.47; CI (mL·s-1·m-2):8.17 ±0.83 vs.48.34 ±3.33,16.84 ±2.17 vs.47.34 ± 1.67,19.00 ± 1.50 vs.47.34 ± 1.33; dp/dt max (mmHg/s):426.83 ± 51.91 vs.1 372.50 ± 39.61,594.00 ± 88.23 vs.1 363.83 ± 44.92,645.00 ±66.82 vs.1 395.83 ± 19.49,all P＜0.05],and the systemic vascular resistance (SVR) and alanine transaminase (ALT),creatinine (Cr),serum MB isoenzyme of creatine kinase (CK-MB),diamine oxidase (DAO) were significantly higher [SVR (kPa·s ·L-1):1 322.50 ±36.37 vs.281.45 ± 8.84,777.50 ±41.84 vs.289.72 ± 6.70,571.40 ±40.01 vs.286.27 ±8.66; ALT (U/L):89.50 ±4.11 vs.40.57 ±3.63,89.25 ±4.88 vs.37.92 ± 2.62,86.30 ±5.61 vs.38.47 ±3.50; Cr (μmol/L):75.62 ±4.61 vs.41.58 ±2.78,77.00 ±5.92 vs.46.55 ± 3.17,74.13 ±2.56 vs.45.65 ± 1.83; CK-MB (kU/L):13.122 ±0.282 vs.1.557 ±0.009,8.885 ±0.272 vs.1.497 ± 0.009,8.692 ± 0.180 vs.1.490 ± 0.005; DAO (kU/L):2.26 ± 0.14 vs.0.25 ± 0.02,1.50 ± 0.07 vs.0.25 ± 0.01,1.37 ± 0.07 vs.0.25 ± 0.02,all P＜0.05].All parameters in NR group kept on worsening till death,while hemodynamic and organ functions of two intravenous resuscitation groups were gradually improved,CI,SVR and DAO in RP group were significantly superior to those of RL group from 2 hours on after burn (all P＜0.05),and dp/dt max and CK-MB in RP group were significantly better than those of RL group from 6 hours on after burn [dp/dt max (mmHg/s):1 082.33 ± 63.59 vs.1 018.60 ± 47.36,CK-MB (U/L):7 898.70 ± 255.74 vs.8 438.70 ± 442.00,all P＜0.05],and MAP (mmHg) was significantly better than that of RL group at 6 hours (124.67 ± 9.39 vs.114.33 ± 9.16,P＜0.05),and Cr (tμmol/L) was significantly better than that of RL group from 24 hours on after burn (53.42 ± 4.99 vs.60.77 ± 3.11,P＜0.05).Conclusion The Pyr Ringer solution was superior to the Ringer lactate solution in improving hemodynamic and organ functions for intravenous resuscitation in dogs with 50％TBSA full thickness burn.

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Objective To explore the effects of PDCA-based self-management intervention model on health behavior and medication adherence in aged patients with percutaneous coronary intervention (PCI). Methods Totally 130 aged patients treated by PCI were randomly divided into the intervention group and the control group with 65 patients. The patients in the control group received routine health education, and the patients in the intervention group received PDCA-based self-management intervention model. All patients were investigated with Coronary Heart Disease Self-Management Behavior Scale (CSMS) and Health Promoting Lifestyle Profile (HPLP) and Medication Compliance Scale (MMAS-8) 3 months and 6 months after discharge. Results Six months after discharge, the score of self-management and healthy behavior and medication adherence were 96.98 ± 14.12, 131.86 ± 16.53, 7.18 ± 0.69 respectively in the intervention group, and the score of them were 86.04 ± 11.78, 105.33 ± 10.97, 5.69 ± 1.29 respectively in the control group, and the difference was statistically significant (t=10.981, 10.793, 7.438, P<0.05 or 0.01). Conclusions PDCA-based self-management intervention model is a patient-centered, problem-oriented, dynamic and interactive health education intervention. It may be helpful in improving PCI patients′ health behavior and medication adherence after discharge. And it may establish lasting self-management skills, and is worthy of application and promotion.

Objective To investigate the effects of recombinant human fibronectin fragment (RetroNectin) combined with anti-CD3 monoclonal antibody (CD3Ab) on the proliferation and cytotoxicity of cytokine-induced killer cells (CIK) from acute leukemia (AL). Methods Mononuclear cells (MNCs) were isolated from peripheral blood of complete remission AL pa-tients. The MNCs were cultured in vitro by precoating with RetroNectin (RN group), CD3Ab (CD3Ab group), RetroNectin com-bined with CD3Ab (RN+CD3Ab group) and traditional method (control group) to generate CIK. The changes of growth rate, characterization, cytotoxicity and apoptosis of CIK were determined between groups. Results The amplification of CIK was higher in experimental group than that of control group, and the amplification of CIK was higher in group RN+CD3Ab than that of in group RN and group CD3Ab (P＜0.05). The expression of CD25 positive cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05).The percentage of G1 stage cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group. The percentage of S stage cells was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05). The cytotoxicity was higher in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P＜0.05) at the E/T scope 40∶1.The percentage of apoptotic cells was lower in group RN and group RN+CD3Ab than that of group CD3Ab and control group (P ＜ 0.05). Conclusion These in vitro studies suggest that a higher activity of immune cells could be obtained by CIK cells cultured by precoating Ret-roNectin and CD3Ab.

Objective To investigate the pathogenic mechanism of Campylobacter jejuni(C.jejuni) associated with Guillain-Barré syndrome(GBS) and provide strategy for gene modification, the cstⅡ gene from 8 GBS-associated C.jejuni strains were compared with that from 3 GBS-unrelated C.jejuni strains, getting the base and amino acid mutations, the changes of secondary structures and finding the region which may be responsible for the pathogenicity of C.jejuni inducing GBS. Methods Three GBS-associated C.jejuni strains isolated from stools of GBS patients in north China were selected and cultured, which has been confirmed as GBS-associated by animal model. After sequencing the genome of them, the nucleotide sequences of cstⅡ gene were got through sequence alignment. The nucleotide sequences and deduced amini acid sequences of 3 GBS-associated cstⅡ genes were compared with that from 3 GBS-unrelated C.jejuni strains through bioinformatics software, getting the base and amino acid mutations, the changes of secondary structures. Other 5 GBS-associated cstⅡ genes were also aligned to know whether the differences we got above makes sense. In this way the genetic differences between two kinds of C.jejuni strains may be found and speculating the gene region related to the pathogenicity of GBS became possible. Results The cstⅡgene of 3 GBS-associated C.jejuni strains were all composed of 876 base pairs. Compared with GBS-unrelated C.jejuni strains, there were 9 consistent mutation sites in cstⅡ gene of LL and QYT stains, leading to 3 consistent amino acid mutation. The amino acid mutation of 114 and 182 sites in LL and QYT stains existed in other 5 GBS-associated C.jejuni strains. The sole amino acid mutation of ZHX strain -169 site, located near the 182 site. The seventh α-helix(165-180 region)of the secondary structure of the amino acid sequence from GBS-associated strains were shorter than that from GBS-unrelated strains, and the shorter regions were opened to form β-sheet or coli, which also existed in other GBS-related strains in this study.75% of the GBS-associated cstⅡ genes were Asn-51, while 25% of the GBS-associated and all of the GBS-unrelated cstⅡ genes were Thr-51.LL strain showed highly identity to other GBS-unrelated strains in this study. Conclusion The 165-180 segment of secondary structures in cstⅡ gene from local 3 GBS-associated C.jejuni strains are probably the responsible region involved in inducing GBS. The senior structure changes in this region may affect the activity of sialyltransferase and the structures of ganglioside epitope, so that the C.jejuni can acquire the pathogenicity of GBS. This finding may give a clue to genetic modified site. The bi-functional cstⅡ of C.jejuni may be related to the pathogenicity of GBS. The cstⅡ of LL strain to some extent represents the characteristics of Asian strains, which may directs strains monitoring.