Practical study on reconstruction of the regional anatomy course system based on clinical practice ability / 中华医学教育探索杂志

Regional anatomy teaching not only requires students to deal with the basic knowledge of human body including the level, location and adjacent relationship, but also to understand the clinical application of anatomical structure. Based on the four aspects of field anatomy, simulated surgery, clinical application lectures and CBL teaching, this study formulated a suitable assessment method to reconstruct the teaching system of regional anatomy relying on the improvement of the laboratory environment and the teacher team, aiming at cultivating students' clinical practice ability as the core and building a new regional anatomy course to meet the teaching needs of the new era.

Electro-acupuncture promotes repair of rotator cuff injury in rats / 南方医科大学学报

OBJECTIVE@#To observe the therapeutic effect of electro-acupuncture on tendon healing and functional recovery of rotator cuff injury in rats and explore the therapeutic mechanism of electro-acupuncture.@*METHODS@#Ninety SD rats were randomly divided into electro-acupuncture group, model group and blank control group, and models of rotator cuff injury were established in the former two groups.The rats in electro-acupuncture group was treated with electro-acupuncture after the operation, and those in the other two groups received no treatment.The right forefoot thermal withdrawal latency (TWL), the contents of IL-1β, IL-6 and TNF-α in the synovial fluid and the maximum tension load of supraspinatus tendon were measured at 2, 4 and 8 weeks after the operation.@*RESULTS@#TWL in the model group was significantly lower than that in the blank control group and electro-acupuncture group at 2, 4 and 8 weeks after the operation (@*CONCLUSIONS@#Electro-acupuncture treatment not only effectively reduces the expression of inflammatory factors to relieve pain, but also promotes the repair of damaged tissue to improve the biomechanical properties of rotator cuff in the rat models.

The compare of stiffness reduction and functional improvements between early passive motion and delayed passive motion in patients undergoing arthroscopic rotator cuff repair / 实用医学杂志

Objective To compare the effects of early passive motion(EPM)and delayed passive motion (DPM)on joint range of motion(ROM),joint function and recurrent tears after arthroscopic rotator cuff repair. Methods A total of 84 cases with middle or large rotator cuff injury were randomly divided into EPM group(43 cases)and DPM group(41 cases). EPM group began shoulder rehabilitation exercise on the first day post-opera-tion and DPM group began 6 weeks later. ROM,pain and function of shoulder joint were evaluated before and 3,6 and 12 months after operation. Results ASES score and UCLA score in EPM group were significantly higher than those in DPM group at 3 months postoperatively(P < 0.05). There was no significant difference in joint ROM,pain and functional between two groups at 6 months and 12 months postoperatively(P > 0.05). There was no statistically significant difference in recurrent tears between two groups at least 12 months follow-up. Conclusion There was no significant difference in functional outcomes,ROM and relative risks of recurrent tears between EPM and DPM in patients undergoing arthroscopic rotator cuff repairs. EPM is conducive to joint ROM and function rapid recovery postoperatively and does not increase the risks of recurrent tears.

Implementation approach discussion for precise medicine based on healthcare big data / 中华医院管理杂志

Based on thorough analysis of the present healthcare big data,and present problems found in precision medicine,the paper proposed the implementation approach of precision medicine driven by healthcare big data.Citing the practice of the hospital as an example,the authors centered on clinical data to build a precision medicine knowledge system,and leveraged in-depth data mining to develop integrated analysis technology for precision medicine.These efforts aim at application development of precision medicine series upon a panoramic medicine knowledge display,so as to enhance medical quality and diagnosis efficiency and translated use of healthcare big data in precision medicine.

Selective lumbar decompression and fusion with pedicle screw fixation for the treatment of degenerative scoliosis / 实用医学杂志

Objective To investigate clinical effect of local decompression and selective interbody fusion with pedicle screw fixation technique for the treatment of degenerative lumbar scoliosis. Methods Medical records of 122 degenerative lumbar scoliosis patients were retrospectively studied. All cases were treated by selective interbody fusion,local decompression and pedicle screw fixation. Group A(L5 fusion)and group B(S1 fusion)un?derwent imaging analysis,and Suk criteria was used to evaluate the fixation,decompression,bone graft fusion. JOA scores were used to evaluate curative effect of excellent and good rate. Results There were no significant difference in operating time,blood loss and postoperative drainage volume between two groups. Follow?up period for all patients is(16.2 ± 0.8)months. X?ray revealed that Cobb′s angle on the coronal plane,the correction rate of lor?dosis and sagittal Cobb′s angle after treatment,the correction rate of lordosis in group B were better than in group A (P0.05). There was no infection or screw breakage and pulling out. The incidence rate of adjacent vertebral body disease in group A was higher than that in group B(P < 0.05). Conclusions Local decompression and selective interbody fusion with pedicle screw fixation technique for the treat?ment of degenerative lumbar scoliosis can produce satisfactory short to mid?term clinical curative effect.

Research on the Induced Differentiation of Adipose Derived Stem Cells and Its Origin / 现代生物医学进展

Objective:To investigate the induction and differentiation potential of ADSCs by tissue culture method,and to preliminary study on the origin of ADSCs.Methods:Using adipose tissue culture method to culture human ADSCs.The third generation of ADSCs for the adipogenic and osteogenesis differentiation,and staining by oil red O and alizarin red S.HE staining was performed after the seventh day culture of adipose tissue.Results:The primary human ADSCs were successfully cultured with adipose tissue culture method.ADSCs cultured to the eighth generation,still maintained a good proliferation ability and cell morphology.ADSCs can be successfully induced into adipose cells and bone cells.ADSCs were mainly distributed around the mesenchymal vascular and connective tissue,by HE staining of adipose tissue after seven days of culture.Conclusion:The cells that were cultured with adipose tissue have the potential to adipogenic and osteogenesis differentiation.The ADSCs were mainly distributed around the mesenchymal vascular and connective tissue.

Effects of Apelin on glucose toxicity and islet cells PDX-1 expression / 重庆医学

Objective To study the effects of Apelin on glucose toxicity and islet cells PDX-1 protein expression.Methods The islet β cell line NIT-1 cells were incubated in the medium containing different glucose concentrations(normal glucose concentration group 5.6 mmol/L,high glucose concentration group 16.7 mmol/L,extremely high glucose concentration group 33.3 mmol/L) and +/-Apelin-36 respectively for 3 d.Then the basic insulin secretion amount of islet cells and their secretion amount after glucose stimulation were detected.The intracellular insulin content and the PDX-1 protein and mRNA expression were detected.Results Compared with the normal glucose group,the basic insulin secretion,secretion after stimulation and intracellular insulin in the high glucose group and extremely high glucose group were significantly decreased and PDX-1 protein expression was declined(P＜ 0.05);compared with non-adding Apelin group,the basic insulin secretion,secretion after stimulation and intracellular insulin in the adding Apelin high glucose group and extremely high glucose group were significantly decreased and PDX-1 protein expression was decreased(P＜0.05);the insulin level in islet cells of 6 groups was positively correlated with PDX-1 protein expression and had no correlation with PDX-1 mRNA expression.Conclusion Apelin may participate in the glucose toxic effect by decreasing PDX-1 protein expression,causes the decrease of insulin secretion,thus plays a role in the pathogenesis of diabetes.

Early efficacy study of matrix-induced autologous chondrocyte implantation repairing knee joint cartilage injury / 中华骨科杂志

Objective To study the feasibility and early efficacy of matrix?induced autologous chondrocyte implantation repairing knee joint cartilage injury. Methods The Matrix?induced autologous chondrocyte implantation was used to repair knee joint cartilage injury in 13 cases (11 males and 2 females) with knee joint cartilage injury from April 2012 to March 2013. The av?erage age was 27.5 years old. All cases were suffering from unilateral focal cartilage defect of knee joint with International Carti?lage Repair Society (ICRS) chondral defect classification system grade III or IV, visual analogue scale (VAS)>3, and all of which had corresponding pain symptoms. The average defect area was 4.2 cm2. Standardized rehabilitation exercise was carried out after matrix?induced autologous chondrocyte implantation. Patients were followed up for 1 years, and knee injury and use osteoarthritis outcome score(KOOS), International Knee Documentation Committee (IKDC), subjective knee form and Lysholm score were col?lected to assess the function. Meanwhile, magnetic resonance observation of cartilage repair tissue (MOCART) score was used to assess the magnetic resonance imaging. Results All patients had been followed?up for 1 year. One patient had meniscus repair under arthroscopy for the meniscus injury caused by downstairs sprain in 6.5 months postoperative, so the score of 12 months post?operative was excluded. The knee range of motion was decreased in 3 months postoperative (123.1°±8.0°) compared to preopera?tive one (135.4°±5.7°), and has no difference in 6 months (136.1°±6.1°) and 12 months postoperative (135.1°±3.6°) compared to preoperative one. The 5 subsets of KOOS score were decreased in 3 months compared to preoperative one, and were significantly increased in 6 months and 12 months. The IKDC has no difference in 3 months (26.1±3.9) compared to preoperative one (43.5± 6.5), and were significantly increased in 6 months (53.3±5.8) and 12 months (62.8±7.2) compared to preoperative one. The magnet?ic resonance observation of cartilage repair tissue (MOCART) score was increased in 12 month(73.3±17.9)compared to preopera?tive one(51.5 ± 12.6). Conclusion MACI is a good technology for knee joint cartilage injury. It has a good clinical effect on re? pairing cartilage injury effectively and restoring the function of knee joint.

Bioinformatics study on human miRNA-Ebola virus interaction / 军事医学

Objective To study the interaction between human microRNA ( miRNA) and 5′trailer regions of Ebola virus genome from the perspective of bioinformatics so as to facilitate prevention and treatment of Ebola virus .Method The miRNA target prediction software Pita and RNAhybrid were used to predict the human miRNAs which could bind the 5′trailer regions of Ebola virus genomes before the miRNAs were annotated by g:Profiler web server .Results and Conclusion There may be complex interactions between human miRNAs and the 5′trailer regions of Ebola virus .Previous reports about the interaction between host miRNA and 5′trailer region of virus genome suggest that the interaction between human miRNA and 5′trailer region of Ebola virus may have effect on replication of Ebola virus and human cells .This work may provide new ideas on prevention and treatment of Ebola virus .

Generation of RNase L knockout cell lines by CRISPR/Cas system / 军事医学

Objective To establish RNase L gene knockout HEK 293 cell lines using CRISPR/Cas9 system.Methods Small guide RNA ( sgRNA) sequences of human RNase L were designed and sgRNAs were inserted into pCas-Guide and pCas-guide RNA(gRNA) vectors were obtained.The donor DNA sequences of the homologous arm were designed for RNase L knockout .In the presence of the right homologous arm , the resistance gene of hygromycin B and the left homologous arm as templates of homology-directed repair , the donor DNA template was amplified by overlopping PCR and cloned into the pBackZero-T expression vector and pBackZero-T-RNase LK vector was obtained .The pCas-gRNA vector and pBackZero-T-RNase LK vector were co-transfected into HEK293 cells to establish the stable expression cell line of RNase L gene knockout .Cells were cultured with hygromycin B , while Western blotting and DNA sequencing were used to analyze the gene of RNase L knockout from genome .Results and Conclusion The pCas-gRNA vector and pBackZero-T-RNase LK vector were successfully constructed.Five RNase L gene knockout HEK293 cell lines were generated,contributing to the study of the biological function and molecular mechanism of RNase L .

Detection of stress-induced 5′tRNA halves by poly(A) tailed-RNase H digestion-RT-PCR / 军事医学

Objective To develop a simple and quick method for detection of stress-induced 5′transfer RNA( tRNA) halves.Methods Total RNA purified from stress induced cells was polyadenylated by poly( A) polymerase, and then degen-erate DNA probes were used to hybridize with 3′tRNA-halves of intact tRNAs,while RNase H specifically degraded the 3′tRNA-halves strand in tRNA-DNA probes hybrids.Using the RNase H digestion total RNA as templates, complementary DNA( cDNA) was synthesized by oligo ( dT) n-anchored primers.The primer of 5′tRNA halves and anchored-primer were used to amplify 5′tRNA halves by PCR.Results The results showed that the method of poly ( A )-tailed-RNase H digestion-RT-PCR could be successfully used to detect stress-induced 5′tRNA halves.Conclusion A simple and quick method for detection of 5′tRNA halves has been established,which is a user-friendly tool for 5′tRNA halves detection and function research.

Establishment and optimization of a method to extract miRNAs from plasma / 军事医学

Objective To develop and optimize a new method to extract miRNAs from plasma.Methods miRNAs were extracted from plasma by mixing it with the extraction solution that contained surfactant and by heating .Then the ribonuclease inhibitor was added into the extraction to prevent RNAs from degradation .The expression level of each miRNA was detected by real-time quantitative PCR in oder to evaluate the feasibility of this method .Results A method which extracted miRNAs from plamsa in just one step was established .The specificity , reproducibility and stability of this method have been demonstrated by real-time quantitative PCR .Conclusion The one-step method is simple , inexpensive , and plasma-saving.It seems like a new method for clinical examination of miRNAs from plasma .

Protein-RNA interactions in Escherichia coli:a genome-wide study / 军事医学

Objective To conduct a pilot study on genome-wide in vivo protein-RNA interactions in E.coli.Methods Bacterial lysate was treated with RNase before the RNA fragments protected by proteins were extracted from treated lysate and used to construct cDNA library that was applied to high-throughput sequencing .Finally, the transcripts bound by proteins were obtained by bioinformatics analysis .Results A total of 3193 transcripts were obtained , including 2234 mRNAs, 47 sRNAs, 39 tRNAs, 11 rRNAs, and 862 intergenic regions .Conclusion Some information of transcripts interacting with proteins in E.coli is acquired , which will facilitate further studies of protein-RNA interactions .

Articular cartilage defect treated with biological osteochondral xenogenic graft transplantation / 中国组织工程研究

BACKGROUND:The methods used to repair articular cartilage defects currently have the cons and pros. Fibrocartilages are commonly used to repair tissues, and the fibrocartilage lacks of the tissue biomechanical properties and chemical properties of normal hyaline cartilage. OBJECTIVE:To investigate the feasibility of biological osteochondral xenogenic graft transplantation to repair articular cartilage defects. METHODS:The normal goats were randomly divided into two groups. The donor pig knee joints were the experimental group. Cylindrical osteochondral with the diameter of 4.5 mm and length of 10 mm were col ected with the Smith&Nephew osteochondral transplantation device, and the patented technology was used for deantigen. The donor goat knee joint osteochondrals were the control group and preserved with cryopreservation. The lesions on femoral trochlea and weight-bearing surface of medial condyle were selected respectively for osteochondral implantation, and the animals were sacrificed at 16 and 32 weeks after operation for the general and pathological section observation. RESULTS AND CONCLUSION:General observation in the experimental showed that the lesions were covered by fibroid tissue;some cartilage of the grafts turned yel ow and there was clear boundary between the surface and the peripheral cartilages;the general and section observation under microscope showed that lesions of the control group were covered by the grafts basical y, and cracks could be seen on the edge of the transplant part. The results show that there is difference between effects of biological osteochondral xenogenic graft transplantation and osteochondral al ograft transplantation for the repairing of articular cartilage defects, and osteochondral al ograft transplantation bas better effect.

Effect of human cytomegalovirus on invasive capability of early pregnant extravillous cytotrophoblasts / 华中科技大学学报（医学）（英德文版）

The effect of human cytomegalovirus (HCMV) on invasive capability of early pregnant extravillous cytotrophoblasts (EVTs) was investigated in vitro. Primary EVTs were obtained by complex phosphoesterasum digestion and gradient centrifugation from villous tissue aseptically taken from healthy pregnant women. Cytokeratin7 (CK7), vimentin (Vim) and c-erbB-2 were immunocytochemically detected to identify source of cells, and HCMVpp65 antigen was assayed to determine the infection state of primary EVTs by immunocytochemical staining. The EVTs were divided into two groups: control group and HCMV group, and the expression of c-erbB-2, matrix metalloproteinase-2 (MMP-2) and MMP-9 proteins was detected in two groups by immunocytochemistry and Western blotting. Enzymic activity changes of MMP-2 and MMP-9 were tested by gelatin zymography in primary EVTs infected with HCMV. The invasion of primary EVTs was detected by cell invasion assay in vitro after they were infected by HCMV. The cell source identification showed that the cells obtained were highly-pure primary EVTs, and primary EVTs could be infected by HCMV. Primary EVTs could express c-erbB-2, MMP-2 and MMP-9 proteins, and as compared with control group, the protein expression was decreased significantly in HCMV groups (P<0.05). Primary EVTs could secrete active MMP-2 and MMP-9 in vitro, and the activity of two MMPs was decreased significantly in HCMV groups (P<0.05). The in vitro cell invasion assay showed that the number of primary EVTs permeating Matrigel in HCMV group was decreased (P<0.05). We are led to conclude that HCMV can infect primary EVTs and inhibit their invasion capability, suggesting that the impaired EVT's invasion capability might be related to the abnormal expression of c-erbB-2, MMP-2 and MMP-9 proteins.

Posterior cruciate ligament reconstruction by double bundle-double tunnel Y-shape of the anterior tibialis tendon allograft / 中华骨科杂志

Objective To investigate the clinical results of posterior cruciate ligament (PCL) reconstruction by double bundle-double tunnel Y-shape of the anterior tibialis tendon allograft. Methods From March 2001 to January 2008, 47 patients underwent PCL reconstruction were included. The allogeneic adult anterior tibialis tendon was prepared into the Y-shape double bundles with the length of 130 mm; A bundle was defined as A-side; B-side was two short bundle (B1, B2 bundle). A bundle was 70 mm in length with a diameter of 10-12 mm. B1 bundle (anterolateral bundle) was 55 mm long with a diameter of 6 mm; B2 bundle(posteromedial bundle) was about 50 mm with a diameter of 6 mm. The allograft ligament was installed through the antero-medial approach. Absorbable interface screws were fixed in the tibial tunnel firstly, and then in the femoral tundles. When being fixed, anterolateral bundle was in flexion of 90°, postero-medial bundle was in 30°. Assisted exercise with knee an angle-locked walking aid had continued for 8-10 weeks. Results The average operating time were 45 min. The average follow-up time was 49.5 months. Preoperative Lachmann was positive in all cases while Lachmann was negative in 39 cases, weakly positive in 5 cases, and positive in 4 cases postoperatively. Post-operative KT-1000 testing, Lysholm score and Tegner activity levels has improved significantly compare with the pre-operative ones. Conclusion The double folded bundles of adult anterior tibialis tendon has sufficient length and diameter for posterior cruciate ligament reconstruction with power tension. The methods of ligament passing through the tunnel has improved to ease the procedure.

A comparative study on arthroscopic posterior cruciate ligament reconstruction using bone-patellar tendon-bone allograft, bone-patellar tendon-bone autograft and semitendinosus tendon autograft / 中国组织工程研究

BACKGROUND: There are many methods for posterior cruciate ligament (PCL) reconstruction, which is involved in many graft materials, but few studies aim to compare the differences in outcomes of different grafts for PCL reconstruction. OBJECTIVE: To compare the clinical results of arthroscopic PLC reconstruction with bone-patellar tendon-bone (B-PT-B) autograft, B-TP-B allograft and semitendinosus tendon autograft. DESIGN, TIME AND SETTING: A retrospective case analysis was completed in the Department of Orthopedics, Guangzhou General Hospital of Guangzhou Area Military Command of Chinese PLA from January 2000 to September 2005. MATERIALS: Totally 76 patients underwent arthroscopic PLC reconstruction from January 2000 to September 2005, with the use of B-TP-B autograft in 21 patients, B-TP-B allograft in 27 patients, semitendinosus tendon autograft in 28 patients. METHODS: A retrospective analysis was performed in 76 patients underwent arthroscopic PCL reconstruction, with the use of B-TP-B autograft in 21 patients, B-TP-B allograft in 27 patients, semitendinosus tendon autograft in 28 patients. Postoperative body temperature was examined duration hospitalization. The follow-up parameters included International Knee Documentation Committee (IKDC) scores, Lysholm knee joint scores, and KT-1000 evaluation.MAIN OUTCOME MEASURES: ①Range of motion. ②joint stability: posterior draw test and KT-1000 test. ③overall function of knee: IKDC scores and Lysholm scores; ④complications and side effect. RESULTS: The time of follow-up visit was 26-79 months. Differences were no statistically significant among the IKDC scores, Lysholm scores, KT-1000 side-side difference, the positive rate of posterior draw test in three groups of patients with PCL reconstruction using B-TP-B autograft, B-TP-B allograft and semitendinosus tendon graft (P > 0.05); 10° flexion limitation was found in 3 cases of B-TP-B autograft, 5° flexion limitation in 1 case of B-TP-B allograft and flexion limitation in 2 case of semitendinosus tendon graft. There was no significant difference in the ratio of knee joint flexion limitation among three groups. No synarthrophysis, wound infection, implant disrupture, screw loose, patellar fracture or vascular nerve injury was observed in three groups of patients; There were 12 cases presenting anterior knee pain in the B-TP-B autograft group and 5 cases presenting posterior knee pain in the semitendinosus tendon graft group. The difference of peal-knee pain incidence was statistically significant among three groups (P=0), the highest in B-TP-B autograft group, then semitendinosus tendon graft group and the lowest in B-TP-B allograft group. The time of post-operative fever in B-TP-B autograft group was earlier than that in the B-TP-B allograft and semitendinosus tendon graft groups (P=0). There was no significant difference between allogreft group and semitendinosus tendon autograft group (P=0.844). The rejections appeared in 4 cases of B-TP-B allograft with the manifestations of the sustained jam-like liquid outflow from tibial tunnel. After dressing, hormones or indomethacln, the rejection was healed. CONCLUSION: The arthroscopic B-TP-B autograft, B-TP-B autograft and semitendinosus tendon autograft have the same clinical curative effect in PCL reconstruction.

Methods in Analyzing Abdominal Fat of Obese Children and Adolescents / 临床儿科杂志

Objectives To assess the clinical value of ultrasonography (US) and bioelectrical impedance analysis (BIA) in analyzing abdominal fat contents of obese children and adolescents through comparison with MRI. A correlation with other obese related metabolic parameters was conducted. Methods Ninety 7-17-y-old obese children and adolescents (60 boys and 30 girls with mean age of 9.6 ± 2.9 y and mean BMI of 24.5 ± 4.5 kg/m2) were recruited. Metabolic parameters were measured, and insulin resistance was estimated according to homeostasis model assess-ment (HOMA-IR). On the same day abdomen subcutaneous fat thickness (SFTUS) was measured by US. Body fat mass (FMBIA) and abdominal visceral fat area (VFABIA) were analyzed by bioelectrical impedance analysis (BIA). After obtaining informed consent, abdominal MRI was performed in 20 subjects. Each section of umbilicus level was analyzed by image threshold value segmentation using SigmaScan Pro 5 and abdominal subcutaneous fat area (SFAMRI) and visceral fat area (VFAMRI) were calculated. Results (1) A strong positive association was found between SFTUS and SFAMRI (P< 0.05), VFABIA and VFAMRI (P < 0.01) respectively. (2) FMBIA and SFAMRI, VFAMRI, SFTUS also showed significant correlations (P < 0.05). (3) VFAMRI showed extremely significant positive correlations with TG, Insulin,C-peptide and HOMA-IR (P < 0.01 ) ; SFAMRI was also correlated positively with them (P < 0.05). (4) SFTUS was correlated positively with UA (uric acid), Insulin, 2HIns (insulin measured at 2 hours after meal), C-peptide,2HC-peptide (C-peptide measured at 2 hours after meal) and HOMA-IR (P < 0.01). (5) VFABIA was correlated significant positively with UA, insulin, TG, 2HIns and HOMA-IR. FMBIA showed positive correlation with UA, Insulin,2HIns, C-peptide, 2HC-peptide and HOMA-IR. Conclusions abdominal subcutaneous and visceral fat of obese children and adolescents evaluated by US and BIA are correlated well with those assessed by MRI, and also correlated well with TG, insulin, C-peptide and other metabolic biochemical parameters. Our data support the value of using cost effective, simple and convenient methods such as BIA and US to evaluate the obese and related metabolic risk of children and adolescents in clinical practice.

An experimental study of telomerase reverse transcriptase (hTERT) DNAzyme on growth of A549/DDP cells / 中国肺癌杂志

<p><b>BACKGROUND</b>Telomerase expresses in many cancers and may contribute to drug-resistance. The aim of this study is to observe the effect of telomerase reverse transcriptase (hTERT) DNAzyme on growth of A549/DDP cells and to explore the possibility of telomerase as a new target in treatment of drug resistance for lung cancer.</p><p><b>METHODS</b>An hTERT DNAzyme was composed. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) modified from Kim's method. MTT was used to show the influence of hTERT DNAzyme and cisplatin on A549/DDP cells.</p><p><b>RESULTS</b>The telomerase activity of A549/DDP cells was down-regulated by hTERT DNAzyme in a dose-dependent manner. The growth inhibition rate of A549/DDP cells was 32.9% by hTERT DNAzyme of 0.25μmol/L, and 60.5% by hTERT DNAzyme combined with 3mg/L cisplatin. The CDI of hTERT DNAzyme and cisplatin was 0.9.</p><p><b>CONCLUSIONS</b>hTERT DNAzyme can inhibit the growth of A549/DDP cells and has a synergistic effect with cisplatin. It is suggested that telomerase may be a new target in treatment of drug-resistant lung cancer cells.</p>

Early immunologic status of patients following hand allotransplantation / 中国组织工程研究

BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.