Anatomical analysis of C 7 pedicle screw implantation based on three-dimensional CT reconstruction model / 中华创伤杂志

Objective:To analyze the ideal entry parameters and entry points for C 7 pedicle screws based on three-dimensional CT reconstruction model. Methods:A retrospective case series study was used to analyze the cervical spine CT image data of 50 adult volunteers collected from April 2021 to March 2022 at Taizhou People′s Hospital Affiliated to Nanjing Medical University, including 25 males and 25 females; aged 20-67 years [(43.8±13.5)years]. No significant structural abnormalities were seen in the cervicothorax of all volunteers. First, the 50 CT images were imported into the medical engineering software Mimics 17.0 to reconstruct the three-dimensional C 7 model, and the C 7 pedicle screw diameter (PSD), pedicle screw length (PSL), sagittal angle (SA) and lateral angle (LA) were measured. Then, the anatomical markers, namely the lateral notch and the midpoint of the inferior edge of the C 6 articular process, were used to measure the horizontal distance between the midpoint of the inferior edge of the C 6 articular process and the ideal entry point (line segment A), and the horizontal and vertical distances between the lateral notch and the ideal entry point (line segments B and C). Finally, the lateral notch and the midpoint of the inferior edge of the C 6 articular process were used as markers to observe the distribution of entry points. The values of the above measured parameters were recorded and compared with each other to analyze the differences between different sides and genders. Intra-group correlation coefficients (ICC) were also used to assess intra- and inter-observer agreement. Results:All 100 pedicles from 50 C 7 models were accessed, with the PSD being (6.5±0.7)mm, PSL being (31.8±4.5)mm, SA being (89.8±8.8)°, LA being (31.0±6.7)°, line segment A being 0.9(-0.4, 2.1) mm, line segment B being (5.8±1.7)mm, and line segment C being (3.6±1.5)mm. All ideal entry points were located medial above the lateral notch; moreover, with the midpoint of the inferior edge of the C 6 articular process as a reference, 71 (71%) of the entry points were located laterally and 29 (29%) were located medially. The 12 mm area around the midpoint of the inferior edge of the C 6 articular process was divided into 6 sections, with 47% of the entry points being located within 2 mm lateral to the midpoint, 25% within 2 mm medial to the midpoint, 19% within 2-4 mm lateral to the midpoint, and only 9% within 50% lateral to the midpoint. The measured parameters were not significantly different between the left and right sides (all P>0.05). The PSD, SA, and line segment B in males were (6.8±0.7)mm, (92.0±8.2)°, and (6.3±1.6)mm, which were significantly greater than those in females [(6.2±0.6)mm, (87.5±8.0)°, and (5.3±1.6)mm] (all P<0.01). The remaining parameters were similar between two genders (all P>0.05). All measured parameters had high intra- and inter-observer agreement (ICC: 0.84-0.91), except for line segment A that had moderate intra- and inter-observer agreement (ICC: 0.46-0.63). Conclusions:For C 7 pedicle screw implantation, when the surface of the lateral mass is used as the reference plane, SA is approximately 90°; with reference to the lateral notch, all the ideal entry points are located medially above it; with reference to the midpoint of the inferior edge of the C 6 articular process, approximately 70% of the entry points are located laterally. There are no significant differences in the parameters between the different sides of screw implantation on pedicles. The measured parameters show high intra- and inter-observer agreement, except for line segment A.

Recombinant Human Peroxiredoxin-5 Improves Anti-tumor Immunity by Inducing Macrophage Polarization / 中国生物化学与分子生物学报

Tumor cells can use different strategies to suppress the immune system and disable them for killing tumor cells. Previous studies have shown that recombinant human peroxiredoxin-5 (hPRDX5) can activate the normal anti-tumor immune, so as to control and eliminate the tumor cells, but its exact mechanism of action needs to be studied in depth. The study aimed to investigate whether hPRDX5 exerts its anti-tumor activity by activating or reversing the polarization state of mouse macrophages RAW264. 7 cells. The results of CCK8 showed that different doses of hPRDX5 could significantly enhance the viability of macrophage compared with the control group (P < 0. 001); The results of Nitric oxide (NO) test showed that hPRDX5 significantly enhanced NO secretion levels in RAW264. 7 cells (P < 0. 001); ELISA experiments revealed that hPRDX5 promotes TNF-α (P<0. 01) and IL-6 (P<0. 001) secretion in RAW264. 7 cells; Flow cytometry revealed that hPRDX5 increased the expression of antigen differentiation cluster (CD) 80 (P < 0. 01) and inducible nitric oxide oxide synthase (iNOS) (P < 0. 001) in RAW264. 7 cells, and reduced the expression of CD206 (P < 0. 001) in RAW264. 7 cells induced by tumor conditional culture solution (TCS); Lactate dehydrogenase (LDH) experiments revealed that hPRDX5 can increase the killing activity of mouse macrophages on mouse pancreatic cancer Panc02 cells. hPRDX5 is able to activate mouse macrophage RAW264. 7 cells, promotes its M1-type polarization, reverses M2-type polarization, and exerts antitumor activity through the immune-enhancing effect.

Contrastive study of three different treatments of unstable distal ulna and radius fractures in older children / 中华实用儿科临床杂志

Objective:To compare the clinical effect of manual reduction and plaster fixation, closed reduction and percutaneous K-wire fixation, as well as open reduction plate and elastic intramedullary pin fixation in the treatment of unstable distal ulna and radius fractures in older children.Methods:The clinical data of 110 cases treated in Department of Orthopedics, Children′s Hospital of Nanjing Medical University from January 2014 to December 2017, for unstable fractures of the distal ulna and radius were retrospectively reviewed.There were 73 males, 37 females, aged from 10 to 14 years[(11.6±2.3) years old]. According to the different treatments, the 110 cases were divided into 3 groups which included manual reduction group (48 patients), closed reduction and percutaneous K-wire fixation group (35 patients, K-wire fixation group) and open reduction with plate and elastic stable intramedullary nail fixation group (27 patients, open reduction group). All cases were treated with plaster external fixation.After the fracture healing, the plaster was removed and then exercises were begun.The data of operative time, fracture healing time, displacement rate and wrist function after treatment were recorded in the three groups and compared statistically.Results:There was no significant difference in general data between 3 groups before operation (all P>0.05). For the manual reduction group, K-wire fixation group and open reduction group, the operation time was (20.2±8.4) min, (35.4±12.2) min and (68.5±20.4) min, respectively; the clinical fracture healing time was (42.1±3.8) d, (44.2±4.3)d and (48.4±5.3) d; the cost of treatment was (2 000.4±551.3) Yuan, (8 000.2±1 151.6) Yuan and (20 010.7±2 453.3) Yuan, respectively.There were significant differences in operation time and cost treatment among the 3 groups ( χ2=11.266, F=58.427, all P<0.05). The Berton score of excellent and good were 89.6% (43/48 cases), 91.4% (32/35 cases) and 92.6% (25/27 cases) 3 months later after surgery in the manual reduction group, K-wire fixation group and open reduction group.The mean follow-up was (11.2±3.7) months(range 6-18 months). In the manual reduction group, there were 6 cases with fracture displacement within 2 weeks after the manual reduction.There were 3 cases with symptoms of caudal irritation in K-wire fixation group, which happened in 1 case in the open reduction group.No complications such as nonunion and iatrogenic nerve injury were found during the follow-ups. Conclusion:There is no significant difference in the wrist function among the 3 methods for the treatment of distal radius and ulna fractures in older children.The manual reduction therapy has a high fracture displacement rate.The open reduction therapy causes maximal operative trauma and costs highly.The percutaneous K-wire method is minimally invasive and has shorter operative time and a lower fracture displacement rate.Therefore, percutaneous K-wire is the optimal treatment for distal ulna and radius fractures.

Research progress in complications following elastic intramedullary nail treatment for long bone fracture of extremities in children / 中华创伤杂志

The elastic stable intramedullary nailing ( ESIN ) technique is widely accepted by pediatric orthopedic surgeons for the following advantages: being minimally invasive, non-destructive on epiphyseal function, easy, rapid postoperative recovery, allowing early functional exercise and having low complication rate. It has become the mainstream technique for the treatment of long bone fractures in children. Although the concept of minimally invasive surgery for children with long bone fractures is constantly evolving and the surgical technique is becoming more and more mature, there are still some complications after ESIN operation, such as tendon and iatrogenic nerve injury, soft tissue irritation and infection, poor alignment after fracture or re-shift, delayed union and non-union, leg-length discrepancy, rotation and angular deformity, osteofascial compartment syndrome, and other clinical problems. These complications often occur due to the negligence of important biological principles, obvious technical errors and the negligence of the characteristics of local anatomy. The authors review the problems of ESIN technique in the treatment of humeral fractures, radius and ulna fractures, femoral fractures and tibial fractures of children, seeking to avoid the complications as much as possible and giving full play to the minimally invasive advantages of ESIN.

Analysis of Curative Efficacy of Different Chemotherapy Regimen Combined with Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation on Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To evaluate the efficacy of autologous peripheral blood hematopoietic stem cell transplantation (auto-PBHSCT) on patients with multiple myeloma( MM) after Sequential different chemotherapy.@*METHODS@#Seven cases of patients with MM were included in the A group, and 14 cases of patients received 4-6 courses of chemotherapy with VAD and MP before transplantation were included in the B group and received 4-6 courses of chemotherapy with VTD and VD before transplantation. Auto-peripheral blood hematopoietic stem cell were mobilized by G-CSF. Condition regimen were melphalan(A group) or bortezomib combined melphalan(B group). IFN-α(A group) or Thalidomide(B group) was used as maintenance treatment after auto-PBHSCT.@*RESULTS@#Two cases of patients reached to complete remission (CR)(2/7,28.6%),1 case got very good partial remission (VGPR) (1/7,14.3%), 4 cases got partial remission(PR) (4/7,57.1%) in A group, and 9 cases got CR (9/14,64.3%), 3 cases got VGPR(3/14,21.4%), and 2 cases got PR(2/14,14.3%) in the B group before auto-PBHSCT. The CR and VGPR were significant difference between 2 groups (P<0.05). All the patients got hematopoietic recovery. In 2 groups, the median time of ANC recovery≥0.5×10/L was 13 (11-16) and 14（11-18）days, that of WBC recovery ≥4.0×10/L were 16(15-19) and 18(16-20)days, Plt recovery ≥ 50 ×10/L was 21 (18-25) and 21(17-25) days. Bone marrow showed CR in 21 to 28 days after transplantation. All of 7 cases of patients remised in 6 to 47 months after transplantation, and 4 cases died lastly and 3 cases failed to be followed up in A group. The median time of progression-free survival(PFS) was 36(6-47) months, and that of overall survival(OS) was 37(7-50) months. In B group, 2 cases of patients remissed in 5 and 17 months after transplantation, and did lastly, 1 case relieved in 12 months after transplantation and failed to be followed up. 1 case of patient relived in 46 months after transplantation, and then received the second auto-PBHSCT, and got CR for 105 months. Other 10 cases got CR, their median time of PFS was 45.5(4-105) months, the median time of overall survival(OS) was 45.5(4-105) months. The PFS and OS were very significant different between 2 groups (P<0.01).@*CONCLUSION@#Bortezomib-based chemotherapy, Auto-PBHSCT and maintenance treatment with thalidomide were favorable to the patients of MM for survival prolongation.

Retrospective Analysis of Genetics Abnormalities in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the characteristics of cytogenetics and molecular genetics in patients with multiple myeloma(MM).@*METHODS@#Fluorescence in situ hybridization(FISH) was used for molecular genetics analysis in 86 cases of newly diagnosed MM, at the same time the chromosome karyotype analysis was performed in 20 cases. Specimen were bone marrow cells.@*RESULTS@#FISH detection showed that 68 cases of MM (79.07%) had at least one type of the molecular genetic abnormalities. The positive rates of IgH rearrangement, 1q21 amplification, D13S319 deletion, RB1 deletion and.P53 deletion were 62.79%, 26.74%, 24.42% ,13.95% and 1.16%, respectively. The positive rate of IgH was significantly higher than that of any other probes(P<0.01). The positive rate of IgH was 79.41% in 68 cases. Out of which the positive rate of IgH single and combined with 1, 2, 3, 4 probes was 59.26%, 24.07%, 11.11%, 5.56% and 0 respectively. The positive rate of IgH only was very signficantly higher than that of combined with any other probes(P<0.01).The positive rate of 1q21 was 33.82% in 68 cases, Out of which the positive rates of 1q21 or combined with 1,2,3,4 probes was 21.74%, 43.48%, 21.74%,13.04% and 0 respectively, the 1q21 probe showed positive as combined with other probes(P<0.01), especially with IgH(P<0.05). The positive rates of D13S319 were 30.88% in 68 cases of patients, out of which the positive rates of D13S319 single or combined with 1, 2, 3, 4 probes was 14.29%, 28.57%, 42.86%, 14.29% and 0 respectively, the D13S319 combined with other probes appeared more significant positive(P<0.01), especially with 1 or 2 probes (P< 0.01). The positive rate of RB1 was 17.65% in 68 cases, the positive rate of RB1 singl or combined with 1, 2, 3, 4 probes were 0, 25%, 50%, 25% and 0, the RB1 appeared positive always combined with other probes, especially with D13S319 probe (P<0.01). The positive rate of P53 was 1.47%, as combined with RB1 and D13S319 probes. The chromosomal karyotyping showed that 3 cases carried abnormal chromosomal and 17 cases carried normal chromosome, Out of which 17 cases showed positive by FISH. There was a significant difference of sensitivity between FISH combined with chromosome karvotyping and single chromosome karvotype (P< 0.01).@*CONCLUSION@#The genetic abnormalies display obvious heterogenicity in MM. The sensitivity of FISH is higher than that of chromosomal karvotyping. If FISH and chromosome karvotyping are combined, the positive rate of abnormality can be raised.

Complications of 2 133 cases of pediatric long bone fracture undergoing elastic stable intramedullary nailing in a single medical center / 中华外科杂志

Objective@#To summarize the clinical data of elastic stable intramedullary nailing(ESIN) in the treatment of long bone fracture of children in a single medical center, and to analyze the problems occurred after the ESIN surgery and corresponding solutions.@*Methods@#A retrospective analysis was conducted regarding the clinical data of 2 133 pediatric long bone fractures conforming to inclusion and exclusion criteria from June, 2005 to December, 2017 in Department of Orthopedics, Children′s Hospital of Nanjing Medical University.There were 1 191 boys and 942 girls, aged from 23 months to 14 years with mean age of (5.7 ± 3.1)years.There were 1 866 cases treated with closed reduction with ESIN, while 267 cases were treated with small incision assisted reduction with ESIN.Postoperative problems have been statistically analyzed.@*Results@#There were altogether 2 133 children, including 603 cases of femur, 311 cases of tibia, 8 cases of fibula, 219 cases of humerus, and 992 cases of ulna/radius.The postoperative complications mainly consist of 62 cases of needle tail irritation reaction, 21 cases of misalignments of fracture alignment, 11 cases of intramedullary nail deformity or angular deformity, 7 cases of limb shortening, 14 cases of limited joint activity, 4 cases of nerve injury, 2 cases of tendon injury, 14 cases of difficult nail removal, 4 cases of cortical cleavage, 8 cases of delayed union, 1 case of nonunion, 6 cases of varus/valgus deformity, 5 cases of epiphyseal injury, 6 cases of ESIN exposure, and 2 cases of metal debris of ESIN′ end.@*Conclusions@#The complications of treatment for children with long bone fractures by ESIN cannot be ignored.To master the important biomechanical properties, to get familiar with the local anatomy and to avoid obvious technical errors can reduce the occurrence of postoperative complications.

Correlation between heart rate variability and severity of coronary artery lesion in patients with coro -nary heart disease / 心血管康复医学杂志

Objective :To study correlation between heart rate variability (HRV) and severity of coronary artery le-sion in patients with coronary heart disease (CHD) and its significance .Methods :A total of 171 CHD inpatients from our department of cardiology were selected .All CHD patients were diagnosed by coronary angiography or cor- onary CT .Severity of coronary artery lesion was judged by Gensini scoring system .According to coronary stenotic degree ,patients were divided into coronary no stenosis group (n＝30) ,mild group (stenosis＜50%,n＝51) ,medi-um group (stenosis 50% ～75%,n＝46) and severe group (stenosis＞75%,n＝44).The 24h ambulatory ECG moni-toring was performed in all patients ,HRV indexes were compared among all groups and their correlation with sever-ity of coronary lesion degree were analyzed .Results :There were significant difference in standard deviation of nor-mal to normal RR intervals calculated over the 24 h period (SDNN) ,standard deviation of normal to normal RR in-tervals in all 5 min segments of the entire recording (SDANN) ,root-mean square of differences between successive normal to normal intervals (rMSSD) and adjacent normal RR interval difference ＞ 50ms stroke accounted for a per-centage of 24h total RR interval (PNN50) among four groups ,and HRV indexes significantly reduced along with coronary lesion aggravated , P＜0.05 or ＜0.01. Multi-factor Logistic regression analysis indicated that HRV inde-xes were independent risk factors for coronary artery lesion aggravation (OR＝1.359～3.254 ,P＜0.05 or ＜0.01). Spearman correlation analysis indicated that SDNN ,SDANN ,rMSSD and PNN50 were significant inversely correla-ted with Gensini score (r＝ -0.510～ -0.271 ,P＜0.05 all).Conclusion : HRV possesses certain clinical application value assessing coronary lesion severity in CHD patients .

Changes of IL-21 and Its Mediated JAK/STAT Signaling Pathway in Patients with Immune Thrombocytopenia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the correlation between JAK/STAT signaling pathways and pathogenesis of immune thrombocytopenia(ITP).</p><p><b>METHODS</b>Twenty-six newly-diagnosed ITP patients was included in this study. They all meet the clinical and hematological criteria for the diagnosis of ITP, and patients with coronary heart disease, severe refractory hypertension, diabetes or with severe liver or kidney function incompetence were ruled out. 24 healthy control without autoimmune diseases, viral infectious diseases and with normal liver and kidney functions were also included. The expressions of Jak3, p-Jak3 mRNA, Stat3, and p-Stat3 were tested and the changes in levels of IL-21 mRNA, IL-21 cell secretion after DEX intervention and AG490 blockade were measured.</p><p><b>RESULTS</b>Compared with the healthy control, patients with ITP had significantly high expressions of Jak3, p-Jak3 mRNA, Stat3 and p-Stat3 protein, which significantly reduced after AG490 blocking (P<0.01). The expression of IL-21 mRNA and the secretion of IL-21 obviously decreased after DEX intervention, but increased after AG490 blocking(P<0.01).</p><p><b>CONCLUSION</b>The pathogenesis of ITP associates with the activation of JAK/STAT signaling pathways, and IL-21-mediated JAK/STAT signaling pathways play regulatory role in ITP.</p>

Percutaneous haemostat stroke-poking reduction and elastic intramedullary nailing for pediatric radial neck fractures / 中华创伤骨科杂志

Objective To evaluate the clinical efficacy of percutaneous haemostat stroke-poking reduction and elastic intramedullary nailing for pediatric radial neck fractures of O'Brien types Ⅱ & Ⅲ.Methods From January 2014 to June 2016,38 children (23 boys and 15 girls) with radial neck fracture of O'Brien type u or Ⅲ were treated by percutaneous haemostat stroke-poking reduction and elastic intramedullary nailing in our department.We had 26 left and 12 right sides.Their ages ranged from 5 to 14 years,averaging 8.6 years.All fractures were fresh.According to O'Brien classification,22 cases were type Ⅱ and 16 type Ⅲ.The operation time,frequency of intraoperative C-arm fluoroscopy,frequency of percutaneous haemostat stroke-poking reduction,and union time were recorded.The elbow function was assessed one day before removal of internal implants according to the Métaizeau scoring system.Results All operations succeeded,lasting from 12 to 25 min (average,16.4 min).The frequency of intraoperative fluoroscopy ranged from 3 to 11 times (average,6.4 times);the frequency of intraoperative percutaneous reduction ranged from 1 to 4 times (average,2.3 times).The patients were followed up for 6 to 22 months (average,1 1.2 months).Postoperative X-ray films showed satisfactory alignment of the fracture ends.All fractures demonstrated clinical and radiographic evidence of complete healing after a mean time of 58 days (from 38 to 72 days).The Métaizeau scoring showed 33 excellent,4 good and one fair cases,yielding an excellent to good rate of 97.4％.Follow-ups revealed no infection,implants breakage,nonunion,fracture redisplacement,or iatrogenic radial nerve injury.Conclusions As percutaneous haemostat stroke-poking reduction can increase the probability of successful reduction at first attempt,reduce frequency of close reduction and X-ray exposure for both children and medical staff,and shorten operation time,the procedure is effective,simple,reliable and minimally invasive,leading to fewer complications.

Genome-wide screening of predicted sugar transporters in Neurospora crassa and the application in hexose fermentation by Saccharomyces cerevisiae / 生物工程学报

The lignocellulolytic filamentous fungus Neurospora crassa is able to assimilate various mono- and oligo-saccharides. However, more than half of predicted sugar transporters in the genome are still waiting for functional elucidation. In this study, system analysis of substrate spectra of predicted sugar transporters in N. crassa was performed at genome-wide level. NCU01868 and NCU08152 have the capability of uptaking various hexose, which are named as NcHXT-1 and NcHXT-2 respectively. Their transport activities for glucose were further confirmed by fluorescence resonance energy transfer analysis. Over-expression of either NcHXT-1 or NcHXT-2 in the null-hexose-transporter yeast EBY.VW4000 restored the growth and ethanol fermentation under submerged fermentation with glucose, galactose, or mannose as the sole carbon source. NcHXT-1/-2 homologues were found in a variety of cellulolytic fungi. Functional identification of two filamentous fungal-conserved hexose transporters NcHXT-1/-2 via genome scanning would represent novel targets for ongoing efforts in engineering cellulolytic fungi and hexose fermentation in yeast.

Application of mathematical algorithm for calculating changes of optic and retina function in mice model of glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma. METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed. RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina. CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

Relationship between Gene Polymorphism of HLA-A(*)/-B(*)-DRB1(*) and Aplastic Anemia in Chinese Han Population of Northwestern Plateau / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between the HLA genes and pathogenesis of aplastic anemia (AA), so as to find the susceptible AA genes.</p><p><b>METHODS</b>Polymerase chain reaction with specific sequence primers (PCR-SSP) method was used to detect the HLA typing of 50 AA patients and 183 normal healthy individuals as controls in Chinese Han population of northwestern plateau.</p><p><b>RESULTS</b>The frequency of HLA-A* 0201 (45.0%), B* 1501 (11.0%), B* 5501 (9.0%) and DRB1* 0901 (19.0%) gene frequences in AA patients were significantly higher than those in controls (Odds Ratio: OR=1.657, 2.138, 2.314 and 1.932, x2=4.882, 3.876, 3.863 and 4.473 (P<0.05). In contrast, A* 0301 gene frequency (4.0%) in AA was significantly lower than that in controls, OR=0.349, x2=4.154 (P<0.05). The male HLA-A* 0201 gene frequency was lower than that in female (38.2% vs 59.4%), and the difference was statistically significant (P<0.05). Concludsion: The HLA-A* 0201, B* 1501, B* 5501 and DRB1* 0901 genes may be considered as the risk markers while A* 0301 gene as a protective marker of AA, the HLA-A* 0201 also shows the sex differences.</p>

Clinical Analysis of Adoptive Immunotherapy after Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation in B Lymphocyte Malignant Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy of autologous peripheral blood hematopoietic stem cell transplantation(auto-PBHSCT) combined with adoptive immunotherapy for patients with B lymphocyte malignant lymphoma(ML).</p><p><b>METHODS</b>A total of 110 cases of ML treated with adoptive immunotherapy after auto-PBHSCT from January 2000 to December 2009 were enrolled in adoptive immunotherapy group (treated group), while 74 cases of ML treated without adoptive immunotherapy after auto-PBHSCT from January 1995 to December 1999 were used as control group. The efficacy of 2 groups were analyzed and compared, 110 case of ML in treated group included 78 cases of non-Hodgkin's lymphoma(NHL), 32 cases of Hodgkin's lymphoma(HL),74 cases of ML in control group included 52 NHL and 22 HL. All of the patients were treated sequentially with chemotherapy regimens for 6 courses. After that, all the patients received auto-PBHSCT. After hematopoietic reconstruction, the patients in treated group were given 6 courses of adoptive immunotherapy(rhIL-2 100 WU/day for 10 days monthly for each course), while the patients in control group were not given immunotherapy. All the patients were followed-up for more than 5 years.</p><p><b>RESULTS</b>There was one patient in each group, who died of liver failure and cerebral hemorrhage respectively within 3 and 2 months, and all the other patients achieved hematopoietic reconstruction. Following-up for 1, 3, 5 years, the disease-free survival (DFS) rate in treated group was 97.3%,93.6%,87.3% while 91.9%, 73.0%, 64.9% in control group. Following-up for 3 and 5 years, there was very significant difference in DFS between 2 groups(P<0.01). The 1,3 and 5 year DFS rate of patients in stage I/II and III/IV in the treated group were 100%,100%,91.7% and 96.5%,91.9%,86.0% respectively while DFS of control group was 100%, 93.3%, 86.7% and 89.8%, 67.8%, 59.3%, there was a significant difference in 3 and 5 years DFS of III/IV stage patients between 2 groups (P<0.01). The 1,3 and 5 year DFS rate of HL patients were 100%, 93.8%,84.4% in treated group and 100%,72.7%,59.1% in control group respectively. There was significant difference in 3 and 5 years DFS of HL between 2 groups (P<0.05). The 1,3 and 5 year DFS rate of stage I/II HL patients were 100%,100%,88.9% in treated group and 100%,100%,80.0% in control group. The 1,3 and 5 year DFS of HL patients in stage III/IV was 100%,91.3%,82.6% and 94.1%,64.7%,52.9% respectively. There was significant difference in 3 and 5 years DFS of III/IV stage of HL patients between 2 groups (P<0.05). The 1,3 and 5 year DFS rate of NHL patients is 96.2%, 93.6%,88.5% in treated group and 90.4%,73.1%,65.4% in control group respectively. There was a significant difference in 3 and 5 years DFS of NHL between 2 groups(P<0.01). The 1,3 and 5 year DFS rate of stage I/II NHL patients was 100%, 100%, 93.3.9% in treated group and 100%, 90%, 90.0% in control group, respectively. The 1,3 and 5 year DFS of NHL patients in stage III/IV is 95.2%, 92.1%,87.3% and 88.1%,69.0%, 59.5% respectively. There was significant difference in 3 and 5 years DFS of III/IV stage NHL patients between 2 groups (P<0.05).</p><p><b>CONCLUSION</b>Therapeutic efficacy is satisfactory for the patients of B lymphocyte ML treated with adoptive immunotherapy after auto-PBHSCT, especially benefited the patients of stage III/IV significantly.</p>

Effects of the interleukin-21 expression in patients with immune thrombocytopenia and its regulation by high-dose dexamethasone / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of immunologic thrombocytopenia(ITP) pathogenesis with the abnormal expression of IL-21, and to explore the association of high-dose dexamethasone (HD-DEX) treatment with the IL-21 expression.</p><p><b>METHODS</b>26 newly diagnosed ITP patients and 24 healthy controls were enrolled in this study. The mononuclear cells and serum were obtain from density gradient centrifugation in the newly diagnosed ITP patients before HD-DXM treatment, and the samples of healthy controls were also used for assays. The protein and mRNA expression of IL-21 on peripheral blood mononuclear cells(MNC) were determined by flow cytometry and real-time reverse-transcription polymerase chain reaction. Plasma levels of IL-21, IFN-γ and IL-4 were determined by enzyme-linked immunoabsorbent assay (ELISA).</p><p><b>RESULTS</b>IL-21 expression on mononuclear cells was significantly higher in ITP patients (13.07%) than that in normal controls (8.2%), the ratio of IL-21/GAPDH mRNA expression on MNC was significantly higher in ITP patients (9.524±0.97) than that in normal controls (3.701±0.60, P<0.01). After HD-DXM therapy, the ratio of IL-21/GAPDH mRNA decreased significantly (5.87±1.21) as compared with the level before treatment. Significantly high levels of serum IL-21, IFN-γ and lower IL-4 were found in ITP patients, as compared with healthy controls. Serum IL-21 and IFN-γ levels in ITP patients decreased significantly after HD-DXM administration (P<0.01), while post-treatment levels of IL-4 were increased significantly, compared with the levels before treatment (P<0.01).</p><p><b>CONCLUSION</b>Therapeutic effect of DXM on ITP associates with down-regalation of IL-21 expression. The increased expression of IL-21 involves in the pathogenesis of ITP.</p>

Effects of salidroside on proliferation of bone marrow mesenchymal stem cells / 中国实验血液学杂志

This study was aimed to investigate the effect of salidroside on proliferation of bone marrow mesenchymal stem cells (MSC) and their secretion of stem cell factor (SCF). MSC were isolated and amplified in vitro via density gradient centrifugation and adherence screening method. MCS were identified by flow cytometry and osteogenic/adipogenic induction. The effects of salidroside on cell proliferation, cell cycle and the SCF secretion of MSC were detected by flow cytometry. The results showed that the salidroside could induce the proliferation of MSC, peaked at the concentration of 1.5 mg/ml and in a time-dependent manner (in 24 h, 48 h and 72 h). Salidroside at 1.5 mg/ml could more effectively increase the percentage of cells in S and G1/M phase. Co-cultured with salidroside at the concentration of 1.5 mg/ml for 48 h, the SCF and the expression levels of SCF mRNA in co-culture supernatant were both significantly increased (P < 0.01). It is concluded that salidroside in a range of certain concentration can obviously promote the proliferation of MSC and increase the expression and secretion of SCF.

Infection status of HBV, HCV and HIV in voluntary blood donors of Chinese Nanjing area during 2010-2013 / 中国实验血液学杂志

This study was purposed to understand the infection of HBV, HCV, HIV among the voluntary blood donors and the epidemic trend in infectious population in Chinese Nanjing area, and to guide the mobilization and recruitment of blood donors. A total of 199777 whole blood samples of voluntary blood donors were tested by ELISA, the nucleic acid technology (NAT) combined detection (HBV-DNA, HCV-RNA, HIV-RNA) was added for detection of the samples with HBsAg,anti-HCV, anti-HIV at least unilateral negative donors from June 10, 2010 to June 9, 2013 years, and these statistic data were analyzed. Every HIV reactive sample(HIV-antibody and/or HIV-RNA) was sent to be confirmed in the Centers for Disease Control and Prevention in Nanjing. The results showed that the voluntary donors' infection rate of HBsAg, anti-HCV, anti-HIV were 0.45%, 0.28%, 0.11% respectively; NAT positive rate was 0.07%, 32 cases were confirmed with anti-HIV positive, in which 30 cases were male (6 cases were repeated blood donors) and 2 cases were female, 3 cases were unconfirmed, in which 2 cases were males and 1 case was female. The statistical analysis demonstrated that the difference of unqualitative rate of HBsAg, anti-HCV, anti-HIV was statistically significant between the first-time and repeated blood donors.It is concluded that the positive rate of anti-HCV and anti-HIV displayed a declining trend year by year in Nanjing voluntary blood donation population from June 10,2010 to June 9, 2013 years. The unqualitative rate of HBsAg and NAT increased with the age increasing, while that of anti-HCV, anti-HIV decreased with age increasing. The unqualitative rate of the repeated blood donors is far lower than that of the first-time blood donors. The ELISA positive rate of anti-HIV testing in females is higher than that in males, but the confirmed positive rate of male is significantly higher than that of female. Therefore the consulting skills before donating should be improved, concerning the link of recruiting donors, focusing on strengthening the first-time donors' consultation, evaluating and developing the fixed voluntary blood donors, and vigorously popularizing NAT technology in blood screening to improve the blood safety effectively.

Curative efficacy for nasal type extranodal NK/T-cell lymphoma by autologous peripheral blood stem cell transplantation after sequencing chemotherapy and radiotherapy / 中国实验血液学杂志

The purpose of this study was to explore the curative efficacy for nasal type extranodal NK/T-cell lymphoma by autologous peripheral blood stem cell transplantation (APBSCT) after sequencing chemotherapy and radiotherapy. A total 65 cases diagnosed as nasal type extranodal NK/T-cell lymphoma by pathology and immuno-histochemistry were treated with chemotherapy and radiotherapy in our hospital from January of 2000 to December of 2009. They were divided into observation group (34 cases) and transplantation group (31 cases). The 34 cases of observation group were ceased from treatment, the 31 cases in transplantation group received APBSCT after conditioning regimen with TBI combined VEMAC. Autologous peripheral blood stem cells were mobilized with chemotherapy combined rhG-CSF. The patients were followed up for 3-5 years. The results showed that some side-effects such as bone marrow suppression and injure of oral cavity mucosa were found in patients after sequencing chemotherapy and radiotherapy. All patients in transplantation group obtained hematopoietic reconstruction, and there were no any special side effect such as VOD. In transplantation group, the median time of ANC ≥ 0.5×10(9)/L was 14 (11-17) days, median time of WBC count ≥ 4.0×10(9)/L was 17 (16-20) days, median time of Plt count ≥ 50×10(8)/L were 25 (23-28) days. After chemotherapy and radiotherapy, effective rate of treatment was 91.2% in observation group, whereas was 90.3% in transplantation group, there were no obvious difference between two groups (P > 0.05). After following up about 1 year, effective rate of treatment was 76.5% in observation group, whereas was 96.8% in transplantation group, there were obvious difference between two groups (P < 0.05). After following up about 3 years and 5 years the disease-free survival (DFS) was 61.3%, 43.5% and 87.1%, 81.5% in observation group and transplantation group, there was significant difference between two groups (P < 0.05). It is concluded that treatment with APBSCT after sequencing chemotherapy and radiotherapy for nasal type extranodal NK/T-cell lymphoma may increase DFS efficiently.

Autophagy is involved in doxorubicin induced resistance of human myeloma cell line RPMI8226 / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the role of autophagy in doxorubicin (DOX)-induced resistance of human myeloma cell line RPMI8226.</p><p><b>METHODS</b>We established doxorubicin induced resistant subline of myeloma cell line RPMI8226/DOX by drug concentration step-elevation method. Resistant index of DOX was measured by MTT assay. Autophagy of myeloma cell lines RPMI8226/s and RPMI8226/DOX was detected by transmission electron microscopy, immunofluorescence (LC3-FITC) and western blot respectively. Apoptosis of RPMI8226/DOX cells induced by DOX combined with autophagic inhibitor hydroxychloroquine or 3-MA was identified by AnnexinV-FITC/PI double fluorescence dyeing.</p><p><b>RESULTS</b>Resistant index of RPMI8226/DOX was approximately 10.8 fold of that of RPMI8226/S. Electron microscopic studies revealed that most of RPMI8226/DOX cells displayed viable attributes and contained numerous autophagic vacuoles. Fluorescent images of RPMI8226/DOX cells showed a punctuate distribution in LC3 protein. Increased LC3-II protein in RPMI8226/DOX cells was determined by immunoblotting. There were no differences among 8 μmol/L HCQ (3.24±1.08)%, 10 mmol/L 3-MA (2.81±0.80)% or control \[(2.12±1.24)%\] (P>0.05) in terms of AnnexinV-FITC/PI double fluorescence dyeing; Compared with apoptosis of (9.75±2.15)%, (24.36±2.16)% and (40.51±3.14)% of RPMI8226/DOX cells under 2, 4 and 6 μmol/L DOX, apoptosis increased significantly after 24 h incubation under 2, 4 and 6 μmol/L DOX combined with 8 μmol/L HCQ as of \[(16.56±1.89)%, (36.44±2.91)% and (62.68±3.75)%, respectively\], or under 2, 4 and 6 μmol/L DOX combined with 10 mmol/L 3-MA as of \[(15.47±1.85)%, (39.28±3.06)% and (55.46±4.07)%, respectively\] (P<0.05).</p><p><b>CONCLUSION</b>Autophagy was involved in doxorubicin-induced resistance of myeloma cell line RPMI8226, DOX resistance in myeloma cells was reversed partly by autophagy inhibitor hydroxychloroquine or 3-MA, and autophagy may be one of mechanisms for drug resistance.</p>

Roles of gap junctions in proliferation mediated by Hcy in the spontaneous hypertensive rat vascular smooth muscle cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate whether homocysteine (Hcy) participates the proliferation of the spontaneously hypertensive rat(SHR) vascular smooth muscle cell (VSMCs) and the molecular mechanism.</p><p><b>METHODS</b>The rat's arota were removed. The primary SHR VSMCs were isolated and cultured in vitro, then the SHR VSMCs were divided into four groups: (1) control group, (2) Hcy group, (3) 18alpha-glycyrrhetinic acid (GA) group, (4) Hcy + 18alpha-GA group. We detected proliferation of the SHR VSMCs by MTT and flow cytometry. The expression and co-localization of the connexin (Cx) 43 and Cx40 proteins in the SHR VSMCs were deteced by immunofluorescence. The expression of the Cx43 and Cx40 proteins in SHR VSMCs were detected by Western blot. The molecular dye transfer method (scrape dye transfer method) was applied to detect the gap junction function in the SHR VSMCs.</p><p><b>RESULTS</b>(1) The Cx43 and Cx40 proteins expression in the SHR VSMCs were positive, confocal microscopy supported the co-localization of Cx43 and Cx40 in the cytoplasm. (2) The S value deteced by cell cycle and A value detected by MTT in the Hcy group were increased obviously compared with those in the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the S and A value in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (3) The expression of Cx43 and Cx40 proteins in Hcy group were increased compared with the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the expression of Cx43 and Cx40 proteins in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (4) The function of gap junction detected by scrape dye transfer method in the Hcy group were enhanced compared with the control group (P < 0.05), weakened in the 18alpha-GA group (P < 0.05). Compared with the Hcy group,the function of gap junction in the Hcy + 18alpha-GA group was significantly weakened (P < 0.05).</p><p><b>CONCLUSION</b>Hcy can enhance the function of gap junctional to stimulate the proliferation of SHR VSMCs through the expression of Cx43 and Cx40 proteins promoted.</p>