BACKGROUND:Recent development of bioengineering technology and tissue-engineered nerve brings a new hope for the treatment of peripheral nerve injuries, which has gradual y become a research spot. OBJECTIVE:To review the new progress in the repair of peripheral nerve injuries using seed cells, biomaterials and tissue-engineered nerve construction technology. METHODS:PubMed and CNKI were searched by the first authors for articles concerning nerve tissue engineering and repair of peripheral nerve injuries published prior to July 2013. The keywords were“tissue engineering, peripheral nerves, nerve injuries, stem cells, Schwann cells, scaffold, growth factor”in English and Chinese, respectively. The articles published recently or in the authorized journals were preferred in the same field. Final y, 63 articles were included in result analysis. RESULTS AND CONCLUSION:Up to now, there is a great advance in the tissue engineering technology for the repair of peripheral nerve injuries. However, most studies are stil in experimental step. For the clinical application of nerve tissue engineering, some problems to be solved include:(1) source and ethics of seed cells;(2) immunological rejection fol owing cellproliferation and transplantation;(3) stability and oncogenicity of transplanted cells;(4) degradation rate, optimal porosity, tube thickness and shape;(5) repair timing for in vitro tissue-engineered nerve construction;(6) local release and regulation of various neurobiological factors. With the development of science, many patients with nerve injuries can profit from the solve of these problems.

Background Cataract surgery can induce the occurrence of dry eye because of the influence of incision on corneal nerve fibers and the change of corneal regularity derived by surgery.Objective This study was to investigate the change of regularity of anterior corneal surface,tear film and astigmatism caused by the process of manual small-incision cataract surgery (manual SICS) and phacoemulsification (Phaco).Methods A prospective non-randomized controlled study was designed.Fifty eyes of 46 age-related cataract patients were included in this clinical study.Thirty eyes of 30 patients received the manual SICS,and other 20 eyes of 16 patients underwent Phaco.Corneal irregularity measurement (CIM),shape factor (SF) and corneal astigmatism were assessed by the corneal tomography examination,the subjective symptoms were scored and tear film break up time (BUT),Schirmer Ⅰ test (S Ⅰ t) were examined before surgery and 1 day,1 week,1 month after surgery.The examined parameters were compared among different time points and two groups.This study was approved by Ethic Committee of Affiliated Eye Hospital of Wenzhou Medical College,and written informed consent was obtained prior to operation.Results The CIM values were gradually increased with the prolong of time after operation,showing a statistically significant difference dependent on time (Ftime =22.864,P =0.000),and CIM value of 1 day,1 week and 1 month after operation was higher than that of before operation (P＜0.01).However,no significant difference was found in CIM value between two groups in different time points (Fgroup =0.062,P =0.804).There was no significant difference in SF values with time prolong (Ftime =4.600,P =0.428) and different groups (Fgroup =0.009,P =0.925).The BUT was gradually reduced with the time prolongs with the considerably difference (Ftime =39.384,P=0.000).Obviously decreases of BUT values were seen in 1 week and 1 month in comparison with 1 day after operation (both P=0.000).But no significant change was found in BUT between two groups in various time points (Fgroup =0.000,P=0.983).In the manual SICS group,the difference was found in S Ⅰ t values between before operation and 1 week or 1 month after surgery(P＜0.01).Surgery associated astigmatism (SIA) was existed in the operative eyes in various time points after operation.With time lapse,SIA value was declined with a significant difference (Ftime =21.479,P =0.000),the difference also was found in different groups (Fgroup =8.709,P =0.005),and SIA value in the Phaco was significantly lower than that in the manual SICS group (P＜0.01).Conclusions In the present study,in the early stage after manual SICS and Phaco,SIA appears due to the irregular change of the anterior corneal surface increases and stability of tear film worsens.

BACKGROUND: As a metal internal fixation material, magnesium alloy has more unique advantages, such as biodegradability and elastic modulus. The elastic modulus of magnesium alloy is similar to the compact bone,which is enough to avoid stress-shelter effect. However, biological activity represents bonding ability with the bone in the body for fracture fixation materials, which is of great significance for studies on new kinds of magnesium alloys.OBJECTIVE: To observe the microstructure of Mg-Zn-Gd alloys and to analyze the corrosion products on the material surface after immersed in simulation body fluid.METHODS: The Mg-Zn-Gd alloys were manufactured by the method of fusion casting. Scanning electron microscope with spectrometer was applied to observe microstructure and distribution of precipitated phase and corrosion products of Mg-Zn-Gd alloys in simulated body fluid.RESULTS AND CONCLUSION: The Mg-Zn-Gd alloy was composed of α-Mg solid solution containing Gd and Zn elements and eutectic structure. The eutectic structure was almost feathery, oval-shaped, herringbone-shaped and strip-shaped along the grain boundary. The main ingredients of eutectic structure included Mg, Zn and Gd elements.Deposition layer was composed of O, Mg, Ca and P elements on the surface of Mg-Zn-Gd alloy after 72 hours soak in Hank's solution. X-ray diffraction analysis showed that film layer contained Mg(OH)2 phase, which promoted calcium salt deposition and reduced the corrosion rate. So Mg-Zn-Gd alloy can obtain better biological activity.

Objective:To study effects of large dose thymopeptides on T cell subpopulations of patients with malignant tumor under radiotherapy.Methods:Fifty one patients with malignant tumor under radiotherapy were divided into 2 groups with 100 mg and 200 mg thymopeptides respectively.The patients we re given thymopeptides,100 mg/d or 200 mg/d,iv, for 10 days.The positive percent ages of CD4,CD8,CD25 and CD56 in T cells of peripheral blood before and after thymopeptide treatment were determined by flow cytometry.Results:The positive percentages of CD4 and CD25 in T cells of peripheral bl ood after 100 mg/d thymopeptide treatment were significantly higher than those befor e thymopeptide treatment (P<0.05),while those of CD4，CD8，CD25 and CD56 in T cells of peripheral blood after 200 mg/d thymopeptide treatment all increased significantly (P<0.05 or P<0.01). Conclusion:These results suggest that large dose of thymopeptides can increase i mmune function of patients with malignant tumor under radiotherapy,and the curat ive effect of 200 mg/d thymopeptides is better.

OBJECTIVE: To analyze the operative methods and clinical effects of autologous distal tibial cancellous bone graft with periosteum in treating cartilage injury of talus with Hepple III-IV type. METHODS: From June 2014 to August 2017, 25 patients (25 feet) with Hepple III to IV cartilage injury of talus were treated with autogenous ipsilateral distal tibial cancellous bone graft. Including 14 males and 11 females, aged from 18 to 52 years with an average of (38.4±3.1) years; left foot was in 8 cases and right foot was in 17 cases. According to Hepple classification, type III of 9 cases, type IV of 16 cases. The curative effect was assessed by Amercian orthopedic foot and ankle society (AOFAS) and visual analogue scale (VAS) before operation and 12 months after operation. RESULTS: All 25 patients were followed up for 12 to 28 months with an average of (14.2±2.5) months. AOFAS and VAS scores were improved from preoperative 53.02±10.06, 8.02±1.14 to 88.04±7.45, 1.26±1.74 at 12 months after operation (<0.05). According to AOFAS standard, 16 cases got excellent results, 6 good, 3 poor. CONCLUSIONS Autologous distal tibial cancellous bone graft with periosteum is an effective method for Hepple III-IV cartilage injury of talus. It can effectively relieve ankle pain and improve ankle joint function.
Adolescent ; Adult ; Bone Transplantation ; Cancellous Bone ; Cartilage ; Female ; Humans ; Male ; Middle Aged ; Periosteum ; Talus ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

Background Leber hereditary optic neuropathy (LHON)is a common inherited eye disease,which generally affects young adults with bilateral loss of central vision.Mutation frequency of Leber hereditary has not been fully clarified. Objective This study was to investigate the mutation frequency of mitochondrial NDI gene associated with LHON in Chinese population. Methods The proposal of the study was approved by Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject initial of this trial.Eight hundred and ninety-four LHON patients and 134 normal subjects were collected.Genomic DNA was extracted from peripheral blood leukocytes of the all participants.Polymerase chain reaction (PCR) was used to amplify and sequence analysis of the mitochondrial ND1 gene was performed and aligned with revised Cambridge Reference Sequence(rCRS) of mitochondrial DNA.Then mutated gene frequency was screened and analyzed. Results Mutational analysis of mitochondrial ND1 gene in 894 LHON patients revealed the presence of G3316A,T3394C,G3460A,C3497T,G3635A,G3733A,and T4216C.11.19％ LHON patients (100/894 ) were found to be associated with the gene mutations mentioned above,and 3.24％ patients (29/894) showed the co-occurrence of three primary mutations.Mutation frequencies in LHON patients were 2.57％,2.23％,1.45％,3.80％,0.67％,0.11％,0.34％,respectively,and G3316A,T3394C,C3497T and T4216C also were detected in 134 normal controls with the mutation frequencies of 4.48％,2.99％,4.48％ and 1.49％,respectively.Mutation frequency analysis showed an insignificant difference in the mutations of G3316A,T3394C,C3497T and T4216C between LHON patients and normal controls (x2 =0.926,P=0.336;x2 =0.052,P=0.820; x2 =0.142,P=0.707;P=0.129).G3376A,G3496T,G3700A,A4136G,T4160C and C4171A were absent in Chinese LHON patients. Conclusions Mitoehondrial ND1 gene in LHON is a mutational hotspot in Chinese population,11.19％ (100/894)associated with LHON was caused by ND1 gene mutation.G3635A,G3733A may be rare pathological mutation in Chinese population.However,G3316A,T3394C,C3497T and T4216C are insufficient to produce the clinical phenotype,but they may play a synergic role for penetrance and phenotypic manifestation in LHON.

OBJECTIVETo establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).

METHODSAccording to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.

RESULTSThe standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.

CONCLUSIONThe fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; DNA Primers ; DNA Probes ; DNA, Viral ; genetics ; Female ; Herpesvirus 6, Human ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis ; virology ; Reproducibility of Results ; Roseolovirus Infections ; diagnosis ; virology ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo clone and express Streptococcus suis serotype 2 (S. suis 2) sly gene for constructing an foundation on identification of S. suis 2 protective antigen.

METHODSThe sly gene was amplified from S. suis 2 clinical isolate strain 05ZYH33 genome DNA by PCR. The gene fragment was inserted into the expression vector pET-30b(+) to build pET30b-sly. When recombinant vector pET30b-sly was identified by restriction enzyme cutting and DNA sequencing as a correct one, subsequently it was transformed to E. coli Rosetta for expression under IPTG induction. The obtained fusion protein was purified by Ni-NTA affinity chromatography. The immunologic and hemolysis activity of the purified protein was proved through Western blot and hemolysis assay respectively.

RESULTSThe PCR product was around 1500 bp. The gene segment inserted into the recombinant vector was proven to be completely identical with the sly gene sequence in the total genome sequence of S. suis 2. The target protein expressed was up to 30% of the total somatic protein under IPTG induction. The protein purity reached above 80% after purification. The protein could be recognized by human serum infected with S. suis 2 and could dissolve swine erythrocytes with the Hemolytic titer as 256.

CONCLUSIONThe expression vector pET30b-sly was successfully constructed. The target protein could be over-expressed in E. coli and possessed its biological activity after purification.

Animals ; Bacterial Proteins ; genetics ; metabolism ; pharmacology ; Blotting, Western ; Chromatography, Affinity ; Hemolysis ; drug effects ; Humans ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; Streptococcus suis ; genetics ; metabolism ; Swine

Objective To assess the efficacy and safety of monoclonal antibody rituximab combined with cyclophosphamide (CTX) in the treatment of refractory and recurrent autoimmune hemolytic anemia.Methods Seven cases with refractory and recurrent autoimmune hemolytic anemia ( including 1 case of Evans syndrome) were recruited during January,2007 to December,2010.Treatment regimens were as follows:rituximab:375 mg/m2,1 time/week,2-6 courses; CTX:1 g,1/10 d,2-7 courses; combined with intravenous immunoglobulin (IVIG) 5 g,1 time/week,given 1 day after rituximab administration.The efficacy and safety of this regimen were assessed during follow-up.Results All the patients showed good responses (7/7).Six patients achieved complete remission (6/7) and one achieved partial remission ( 1/7 ).Average follow-up time for the patients was 27 months.All patients remained in remission during the 12-month follow-up visits.Two patients showed elevated indirect bilirubin and increased reticulocyte counts within 24 months.One patient achieved complete remission after additional rituximab therapy,and another patient remained partial remission after cyclosporine therapy.At the time of 36-month follow-up visit,the patient relapsed and was retreated with 3 courses of rituximab combined with CTX and eventually achieved partial remission.All patients tolerated the treatment well with few mild side effects.Conclusions Rituximab combined with CTX is effective and relatively safe in patients with refractory and recurrent autoimmune hemolytic anemia.Additional treatment to relapse patients about 12-24 months after drug withdrawal continues to be effective.