OBJECTIVETo estimate the effects of the treatment of 360 degrees fixation of lumbar spine.

METHODSTwenty-five cases were operated to 360 degrees fixation of lumbar spine from May 2002 to May 2005. There were 11 male and 14 female, the age ranging from 45 to 67 years, mean 56, including 16 cases of spondylolysis, 7 cases of lumbar spinal instability and 2 cases of degenerative disc undergone with anterior laparoscopic lumbar interbody fusion with syncage and autograft and posterior decompression with facet fixation. The chief complains were low back pain and radiating to lag. Fourteen cases were fused at L(4), 5 and 11 cases at L(5)-S(1). The Oswestry Disability Index (ODI) were recorded to evaluate the function at pro-operation, the second week, the third, sixth and twelfth month post-operation. The X-ray was taken to observe the fusion and the sinking cage.

RESULTSThe time intro-operation was from 110 to 180 minutes, mean 120 minutes. There was one complication in 1 case by the main vein injury. All of the 25 cases were followed-up post-operation from 12 to 35 months, mean 22.3 months. The ODI was found prominent difference compared with pro-operation in 2 weeks, 3, 6 and 12 months post-operation (P < 0.05). Twenty-five cases were fused in the third month. Two cases sank 1 mm in the third month images but solid fusion. No cage displacement was found. No retrograde ejaculation happened.

CONCLUSIONS360 degrees fixation of lumbar spine adapts to deal with lumbar instability, degenerative disc disease and I, II degree spondylolysis. It shows less bleeding and injury, but during exposure and traction the main vessels should be very careful.

Aged ; Decompression, Surgical ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; methods

OBJECTIVENeonatal lupus erythematosus (NLE) is an uncommon autoimmune disease passively transmitted from the mother in which there is transplacental passage of maternal antibodies. It is often misdiagnosed as intrauterine infection or sepsis. The main purpose of this retrospective study was to improve the understanding of pathogenesis and clinical manifestations of NLE.

METHODSClinical manifestations, results of the tests for antinuclear antibodies (ANA), anti-Ro/SSA, anti-La/SSB and anti-dsDNA antibodies in both infants with NLE (8 cases) and their mothers and head ultrasound and CT scans of the infants were analyzed. Follow-up was performed until one and ahalf years of age or all the abnormalities had been resolved.

RESULTSTotally 8 cases (3 males and 5 females) matched the criteria for diagnosis of NLE from September 2003 to February 2006, among whom 4 were small for gestational age and one was born prematurely. Mean gestational age was 38.1 +/- 1.9 weeks, mean birth weight 2 605 +/- 420 grams, mean admission age 22.4 +/- 27.7 days (2 hours-72 days) and mean age of onset 9.4 +/- 12.1 days (0 - 28 days). The common clinical manifestations included cutaneous lupus lesions (8 infants), neural system abnormalities (2 infants) and congenital heart block (2 infants). The skin of the infants exhibited annular, erythematous or desquamative lesions. They all disappeared before 6 months of age. One patient presented with grade III atrioventricular block and was delivered by cesarean section because of "fetal distress". He did not recover at one and a half years follow-up. One infant was hypotonic with delayed neuro-motor development initially and during follow-up with both abnormal neonatal behavioral neurological assessment (NBNA) and image findings. CT showed generalized low density involving periventricular area and deep white matter at one week of age. At the age of one and a half years, he presented with normal mental development index determined by CDCC infant intelligence mensuration. Other abnormal clinical findings included hepatosplenomegaly, anemia, thrombocytopenia, cholestasis and elevated liver enzymes, which were all resolved before 6 months of age. Only 3 mothers of the NLE infants were diagnosed as lupus erythematosus before parturition and only one received partial therapy. At least anti-Ro/SSA antibody or anti-La/SSB antibody or ANA was found in the affected patients. Seven cases had circulating anti-Ro and/or anti-La antibodies in the mothers and in the newborns, while ANA was positive in 7 newborns and in all mothers. All the clinical symptoms except congenital heart block disappeared before 18 months of age. No special intervention was applied.

CONCLUSIONSerum auto-antibodies should be investigated to rule out NLE when there is congenital heart block or rashes or thrombocytopenia presented in a neonate, despite there is no maternal history. Central nervous system abnormalities in NLE are likely to be transient and whether it will cause long term sequelae is uncertain.

Autoimmune Diseases ; pathology ; Female ; Humans ; Infant ; Infant, Newborn ; Infant, Newborn, Diseases ; diagnosis ; Lupus Erythematosus, Cutaneous ; diagnosis ; pathology ; physiopathology ; Male ; Retrospective Studies

Low birth weight (LBW) and preterm birth (PB) are associated with newborn mortality and diseases in adulthood.We explored factors related to LBW and PB by conducting a population-based case-control study from January 2011 to December 2013 in Wuhan,China.A total of 337 LBW newborn babies,472 PB babies,and 708 babies with normal birth weights and born from term pregnancies were included in this study.Information of newborns and their parents was collected by trained investigators using questionnaires and referring to medical records.Univariate and logistic regression analyses with the stepwise selection method were used to determine the associations of related factors with LBW and PB.Results showed that maternal hypertension (OR=6.78,95％ CI:2.27-20.29,P=0.001),maternal high-risk pregnancy (OR=1.53,95％ CI:1.06-2.21,P=0.022),and maternal fruit intake ≥300 g per day during the first trimester (OR=1.70,95％ CI:1.17-2.45,P=0.005) were associated with LBW.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.48,95％ CI:0.32-0.74,P=0.001) and gestation ≥37 weeks (OR=0.01,95％ CI:0.00-0.02,P＜0.034) were protective factors for LBW.Maternal hypertension (OR=3.36,95％ CI:1.26-8.98,P=0.016),maternal high-risk pregnancy (OR=4.38,95％ CI:3.26-5.88,P＜0.001),maternal meal intake of only twice per day (OR=1.88,95％ CI:1.10-3.20,P=0.021),and mother liking food with lots of aginomoto and salt (OR=1.60,95％ CI:1.02-2.51,P=0.040) were risk factors for PB.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.66,95％ CI:0.47-0.93,P=0.018),distance of house from road ≥36 meters (OR=0.72,95％ CI:0.53-0.97,P=0.028),and living in rural area (OR=0.60,95％ CI:0.37-0.99,P=0.047) were protective factors for PB.Our study demonstrated some risk factors and protective factors for LBW and PB,and provided valuable information for the prevention of the conditions among newborns.

This study was purposed to directly induce murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) by simulating the spatial and temporal hematopoietic microenvironment changes in embryonic development, and to investigate the function of in vivo hematopoietic reconstitution of these HSC. E14 ESC were induced into embryoid body (EB) firstly. Then the cells from EB were further co-cultured with human aorta-gonad-mesonephros (AGM) region, fetal liver (FL) and bone marrow (BM) stromal cells in Transwell non-contact system in sequential orders. After 6 days of each co-cultured stage, the induced cells derived from EB were collected to analyze the Sca-1(+)c-Kit(+) cells by flow cytometry, check teratoma formation and transplant to BALB/C female mice exposed to lethal dose (60)Co γ-ray. The recipient mice were divided randomly into 5 groups: AGM, AGM + FL, AGM + FL + BM, irradiation control and normal control groups. The survival rates, hematopoietic reconstitution and engraftment of donor cells in the different groups were monitored. The results showed that Sca-1(+)c-Kit(+) cell level in EB cells co-cultured with human AGM region and FL stromal cells reached to peak value (21.96 ± 2.54)%. Teratomas could be found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region stromal cells, while there was no teratoma in the mice after subcutaneous injection of EB cells induced by human AGM region and FL stromal cells. The recipients in AGM group and irradiation control group all died. The survival rate was 77.8% in AGM+FL group, and 66.7% in AGM+FL+BM group. The peripheral blood cell count was near normal at day 21 after transplantation, and Sry gene copies from donor could be detected in recipient mice of these two groups. It is concluded that sequentially inductive system with feeder cells from human AGM region, fetal liver and bone marrow simulating embryonic defined hematopoiesis procedures can enhance the directed differentiation of ESC into HSC which can safely reconstitute hematopoiesis in vivo.
Animals ; Aorta ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Female ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesonephros ; Mice ; Mice, Inbred BALB C ; Mice, SCID

This study was aimed to investigate the effect of human aorta-gonad-mesonephros (AGM) region stromal cells on differentiation of murine embryonic stem cells (ESC) into hematopoietic stem cells (HSC) in vitro and to clarify their effect mechanism. E14 murine ESC were induced into embryo body (EB) firstly. Then the EB cells were further co-cultured with the stromal cells from human AGM region, fetal liver (FL) or bone marrow (BM) in Transwell non-contact system. According to the different culture methods, the EB cells were divided into 6 groups including EB control group, AGM group, FL group, BM group, AGM + FL group and AGM + BM group. The induced cells derived from EB were collected for Sca-1(+)/c-Kit(+) cells analysis by flow cytometry and colony forming unit (CFU) assay. The results showed that Sca-1(+)/c-Kit(+) cell proportion of EB cells significantly increased after being induced by different stromal cells (p < 0.01). The AGM + FL group had most Sca-1(+)/c-Kit(+) cells for the positive cell proportion reached (21.96 ± 2.54) % (p < 0.01). The Sca-1(+)/c-Kit(+) cell proportion of AGM + BM group was much high than that of BM group too (p < 0.01). The EB control group showed CFU amount less than any other groups, while the CFU amount of AGM + FL, AGM + BM groups were higher, especially in the AGM + FL group (p < 0.01). It is concluded that the human AGM region stromal cells may help to maintain certain number of primitive HSC with high proliferation potential. Human AGM region, FL or BM stromal cells, applied in sequential orders, can significantly expand in vitro the primitive hematopoietic stem/progenitor cells derived from ESC.
Animals ; Cell Differentiation ; Cell Line ; Coculture Techniques ; Embryonic Stem Cells ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesonephros ; cytology ; Mice ; Stromal Cells ; cytology

When hematopoietic stem cells (HSCs) were administrated by intravenous infusion (IV), most of them were trapped in some nonhematopoietic organs as like lungs that had abundant blood capillaries. Only a small fraction of injected cells could home to the bone marrow, which reduced the engraftment of HSCs. The purpose of intra-bone marrow (IBM) transplantation was to facilitate the homing of HSCs directly. Based on the established murine model for allogeneic umbilical cord blood transplantation (UCBT) by IBM injection, the objective of this study was to compare the distribution of fetal and neonatal peripheral blood (FNPB) mononuclear cells (MNC) in vivo and the efficacy of HSCT by different routes of administration in mice. BALB/c recipient mice exposed to sublethal dose 60Co gamma-ray were transplanted with FNPBMNCs from C57BL/6 mice. Recipient mice were divided into six groups at random: unilateral-IBM group; bilateral-IBM group; IV group; bilateral-IBM + IV group; irradiated control group and normal group. The distribution of CFSE-labeled FNPBMNCs in the recipients was observed in frozen sections of different organs or by flow cytometry. The survival rate, engraftment level, recovery of hematopoietic function and GVHD of recipient mice were studied. The results showed that infused by IBM route, FNPBMNCs mainly accumulated in the bone marrow (BM) cavity of the injected side tibia. Some of them could enter the BM of noninjected bones via blood circulation and few were trapped in the lung. Though same amount of FNPBMNCs were injected into recipient mice of unilateral and bilateral-IBM group, less cells could leak into peripheral blood or other tissues when transplanted by bilateral-IBM route. Therefore, in term of accelerating hemopoietic recovery, the injection of IBM route was better than IV route, especially bilateral IBM injection of HSCs, which neared the normal level of peripheral blood cells and colony-forming units of bone marrow nucleated cells at day 21 after transplantation, followed by unilateral-IBM group and bilateral-IBM + IV group. The percentages of H-2Db cell subsets in the three IBM groups were much higher than that in IV group. There was no significant difference of the engraftment level in the injected side tibia between the unilateral and bilateral-IBM group. When secondary transplantation was performed, the engraftment level in bilateral-IBM group was still much higher than that in IV group. At day 90, the survival rates of IBM groups were all > or = 80%, while that of IV group was only 50%. It is concluded that bilateral-IBM route can facilitate the homing of more HSCs, accelerate the engraftment of HSCs and hematopoietic reconstitution, which promoted the efficacy of IBM-HSCT.
Animals ; Female ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Random Allocation ; Whole-Body Irradiation

OBJECTIVETo observe the in vivo distribution of mesenchymal stem cells (MSCs) after administrated by intra-bone marrow (IBM) or intravenous (i.v.), and compare the effects on hematopoiesis reconstitution and GVHD in rat BMT models.

METHODS(1) MSCs from male Wistar rats marked with CFSE were injected into the bone marrow cavity (IBM) or the vein (i.v.) of recipient rats, and observed the distribution of MSCs in vivo. (2) Allogeneic BMT model of Fischer344 rats (RT1A(1)) to Wistar rats (RT1A(u)) was established. The recipient rats were exposed to 8 Gy of gamma irradiation 1 day before transplantation. The 6 groups were (1) IBM group [IBM-injection of MSCs + IV-injection of bone marrow cells (BMC)]; (2) IV group (i.v.-injection of MSCs (i.v.) + i.v.-injection of BMC); (3) BMT group (only i.v.-injection of BMC); (4) MSCs control group (only i.v.-injection of MSC); (5) normal control group and (6) irradiation control group.

RESULTS(1) After i.v.-injection, large numbers of the MSCs lodged in lungs while small numbers in the peripheral blood, liver, thymus and spleen, and a few marked MSCs could be seen in bone marrow. After IBM injection, most cells distributed in long bones and those lungs were less than that in i.v. group. (2) Co-transplantation of MSCs (IBM/IV) could accelerate the recovery of hematopoiesis, including the recovery of WBC, hemoglobin and platelet, and in IBM-injection was more effective in the recovery of hematopoiesis than that in i.v. group. (3) Incidence rate of GVHD in BMT group was 42% (3/7), and no GVHD occurred in co-transplantation groups. (4) Recovery of CFU-Mix and CFU-MSCs could be seen at 21st and 30th day after transplantation in co-transplantation groups, and IBM-injection was more effective than i.v.-injection.

CONCLUSION(1) IBM-injection results in most MSCs distributed in long bones. (2) MSCs improve the survival rate after BMT. (3) Co-transplantation of MSCs accelerates the recovery of hematopoiesis and reduces the morbidity of GVHD. (4) MSC promotes reconstitution of hematopoietic cells and bone marrow MSCs in recipient rates and the effects of MSCs administrated via IBM is more effective than via i.v.

Animals ; Bone Marrow Transplantation ; Female ; Graft vs Host Disease ; prevention & control ; Hematopoiesis ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Models, Animal ; Rats ; Rats, Wistar ; Transplantation, Homologous

OBJECTIVETo evaluate the clinical and laboratory features of pediatric inv(16) acute myeloid leukemia (AML) retrospectively.

METHODDual color fluorescence in situ hybridization (D-FISH) using a LSI CBFβ inv(16) break apart probe labeled by Spectrum red and Spectrum green was performed in 15 acute myeloid leukemia cases, including 13 cases with or without abnormal eosinophils but with positive core binding factor β (CBFβ)-MYH11 fusion transcript detected by RT-PCR, and 2 cases with trisomy 8 (+8). The results were compared with the morphology, immunophenotype, karyotype and RT-PCR.

RESULTMorphologically, 12 cases were diagnosed as M(4)EO, 2 as M(4), and 1 as M(2a). Immunophenotypically, all 13 AML cases with inv(16) showed positive expression of CD(13) and CD(33), but without the expression of any lymphoid lineage antigens. Karyotyping analysis with G-banding detected inv(16) in 10 AML cases, including 9 M(4)EO cases and 1 M(2a), but only 5 positive cases were detected using R-banding technique. Among them, 2 cases had simultaneous +8 and trisomy22 (+22), one had +22 only in addition to inv(16). D-FISH revealed a CBFβ-MYH11 rearrangement in 13 cases of AML with positive RT-PCR results, and the mean positive rate of cell detection was 55.15% (range 37.0% - 86.0%). The complete remission rate (CR) and median survival period in this series of inv(16) AML were 81.5%and 11 months, respectively, of whom, 8 cases were still in CR. Relapse and karyotypic evolution were seen in case 5 with +8, +22 in addition to inv(16).

CONCLUSIONAML with inv(16) is a special subtype. Most cases belong to M(4)EO. Its prognosis is good in general, but it seems to be an unfavorable feature for AML with inv(16) and +8, +22 simultaneously, especially with karyotypic evolution. For detection of inv(16), G-banding technique is evidently superior to R-banding technique. D-FISH combined with RT-PCR are more sensitive and reliable than chromosome banding analysis.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Eosinophilia ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Prognosis ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction

This study was aimed to investigate the effect of intra-bone marrow (IBM) injection of allogeneic mesenchymal stem cells (MSCs) on reconstruction of bone marrow MSCs (BM-MSCs) in rats that received hematopoietic stem cell transplantation (HSCT), and to detect the donor MSCs in the hosts for clarifying the effect mechanism of donor MSCs. Wistar female rats conditioned with lethal dose 60Co gamma-ray irradiation were co-transplanted with F344 female fetal and neonatal peripheral blood (FNPB) and BrdU-labeled MSCs separated from bone marrow mononuclear cells of F344 male rats. The donor MSCs were infused by IBM injection in bilateral tibia or intravenous injection (IV), while the FNPB were all via IBM route. The survival rate, engraftment level of HSCs and recovery of BM-MSCs of recipients were monitored. The ratio of BrdU-labeled MSCs in recipient rats was calculated by immunofluorescence assay (IFA) and the Y chromosomes were examined by PCR. The results showed that the recipient rats of the two co-transplantation groups were all alive at day 60 after transplantation. There was no significant difference between these two groups on the survival rates or the engraftment levels of HSCs, but each of them was much better than that of the FNPB group. At day 30 after transplantation, the proliferation ability of recipients' BM-MSCs was still below normal, while that of the FNPB (IBM)+MSC (IBM) group was the best of all the experiment groups (p<0.01). At 60 days, the donor MSCs coexisted with host MSCs in only a few recipient rats examined by IFA, while the Y chromosomes could be detected in all the recipient rats in the two cotransplantation groups. It is concluded that the infusion of allogeneic MSCs can accelerate the recovery of HSCT recipients' BM-MSCs. The IBM route is safe and more effective than intravenous infusion.
Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Cells, Cultured ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Inbred F344 ; Rats, Wistar

BACKGROUNDExperimental studies and preliminary clinical studies have suggested that growth hormone (GH) treatment may improve cardiovascular parameters in chronic heart failure (CHF). Recombinant human GH (rhGH) has been delivered by a recombinant protein, by plasmid DNA, and by genetically engineered cells with different pharmacokinetic and physiological properties. The present study aimed to examine a new method for delivery of rhGH using genetically modified bioartificial muscles (BAMs), and investigate whether the rhGH delivered by this technique improves left ventricular (LV) function in rats with CHF.

METHODSPrimary skeletal myoblasts were isolated from several Sprague-Dawley (SD) rats, cultured, purified, and retrovirally transduced to synthesize and secrete human rhGH, and tissue-engineered into implantable BAMs. Ligation of the left coronary artery or sham operation was performed. The rats that underwent ligation were randomly assigned to 2 groups: CHF control group (n = 6) and CHF treatment group (n = 6). The CHF control group received non-rhGH-secreting BAM (GFP-BAMs) transplantation, and the CHF treatment group received rhGH-secreting BAM (GH-BAMs) transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups: sham control group (n = 6) and sham treatment group (n = 6). The sham control group underwent GFP-BAM transplantation, and the sham treatment group underwent GH-BAM transplantation. GH-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats with ligation of the left coronary artery or sham operation was performed. Eight weeks after the treatment, echocardiography was performed. hGH, insulin-like growth factor-1 (IGF-1) and TNF-alpha levels in rat serum were measured by radioimmunoassay and ELISA, and then the rats were killed and ventricular samples were subjected to immunohistochemistry.

RESULTSPrimary rat myoblasts were retrovirally transduced to secrete rhGH and tissue-engineered into implantable BAMs containing parallel arrays of postmitotic myofibers. In vitro, they secreted 1 to 2 microg of bioactive rhGH per day. When implanted into syngeneic rat, GH-BAMs secreted and delivered rhGH. Eight weeks after therapy, LV ejection fraction (EF) and fractional shortening (FS) were significantly higher in CHF rats treated with GH-BAMs than in those treated with GFP-BAMs ((65.0 +/- 6.5)% vs (48.1 +/- 6.8)%, P < 0.05), ((41.3 +/- 7.4)% vs (26.5 +/- 7.1)%, P < 0.05). LV end-diastolic dimension (LVEDD) was significantly lower in CHF rats treated with GH-BAM than in CHF rats treated with GFP-BAM (P < 0.05). The levels of serum GH and IGF-1 were increased significantly in both CHF and sham rats treated with GH-BAM. The level of serum TNF-alpha decreased more significantly in the CHF treatment group than in the CHF control group.

CONCLUSIONSrhGH significantly improves LV function and prevents cardiac remodeling in rats with CHF. Genetically modified tissue-engineered bioartificial muscle provides a method delivering recombinant protein for the treatment of heart failure.

Animals ; Bioartificial Organs ; Echocardiography ; Heart Failure ; therapy ; Human Growth Hormone ; administration & dosage ; Myoblasts, Skeletal ; metabolism ; Myocardial Infarction ; pathology ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; administration & dosage ; Tissue Engineering ; Tumor Necrosis Factor-alpha ; blood ; Ventricular Function, Left