BACKGROUND:Previous studies have mainly focused on costal cartilage, articular cartilage, nasal septal cartilage, and auricular cartilage, but in vitro culture of human vertebral endplate cartilage is stil rarely reported. OBJECTIVE: To discuss the feasibility of culture of vertebral endplate chondrocytes from type I neurofibromatosis associated with scoliosis patientsin vitro and to study the biological characters of the chondrocytes. METHODS: Through two-step enzymatic digestion and tissue culture, the chondrocytes from the vertebral endplate of seven type I neurofibromatosis patients isolated and cultured in monolayer and passaged to observe the changes of cel morphology under inverted phase contrast microscope. Colagen type II expression was detected by immunocytochemistry to identify whether the cels had chondrocyte characters. The growth kinetics was detected by using MTT colorimetric assay to draw the growth curve of passage 2 chondrocytes. RESULTS AND CONCLUSION:A few chondrocytes crawled from the cartilage after 2 weeks culture and cels were passaged at 3 weeks. Along with passage going on, the phenotype of chondrocytes was changed from polygonal, round, triangle, and irregular shapes to fusiform. The colagen type II expression in passage 2 cels was positive by immunohistochemical staining. MTT test showed the growth curve of the passage 2 chondrocytes presented a transverse “S”. Cels were found logarithmic growth at days 4-7, reached platform stage at days 8-13, and decreased at day 14. It is an effective and simple procedure by two-step enzymatic digestion and tissue explant method to culture vertebral endplate chondrocytes with high purity and good viability from type I neurofibromatosis patients associated with scoliosisin vitro. Passage 2 chondrocytes from the vertebral endplate exhibit the best viability at days 4-7, which can be used as targets for research of pathogenesis of type I neurofibromatosis with atrophic scoliosis.

The combined use of chemometrics and chemiluminescence (CL) measurements,with the aid of the stoppedflow mixing technique,developed a simple time-resolved CL method for the simultaneous determination of captopril (CPL) and hydrochlorothiazide (HCT).The stopped-flow technique in a continuous-flow system was employed in this work in order to emphasize the kinetic differences between the two analytes in cerium (Ⅳ)-rhodamine 6G CL system.After the flow was stopped,an initial rise of CL signal was observed for HCT standards,while a direct decay of CL signal was obtained for CPL standards.The mixed CL signal was monitored and recorded on the whole process of continuousflow/stopped-flow,and the obtained data were processed by the chemometric approach of artificial neural network.The relative prediction error (RPE) of CPL and HCT was 5.9％ and 8.7％,respectively.The recoveries of CPL and HCT in tablets were found to fall in the range between 95％ and 106％.The proposed method was successfully applied to the simultaneous determination of CPL and HCT in a compound pharmaceutical formulation.

This paper was aimed to study the genetic diversity and genetic relationship of germplasm resource of Lonicerae Japonicae Flos in order to provide references for its breeding. A total of 36 samples from 18 farm varieties and wild species, as well as related species of Lonicera japonica Thunb. from the main production areas were studied by ISSR-PCR markers. The Jaccard coefficient was worked out by NTSYS-pc software. And a cluster dendrogram of different samples was established based on unweighted pair-group method with arithmetic mean (UPGMA). The re-sults showed that 12 ISSR primers generated 129 loci of which 114 loci were polymorphic. The average percentage of polymorphic bands (PPB) is 88.37%. In the cluster dendrogram, different samples of Lonicera japonica are in the same group, which showed that it is a natural species; the wild sample is separated from the cultivated samples; the traditional type-Maohua is relative stable, but the type of Jizhuahua includes complicated varieties, and it has obvi-ous genetic variation; the new variety Jiufeng 1 has already distinct into one stable type. It was concluded that the ISSR method was suitable for germplasm identification, genetic diversity analysis of Lonicerae Japonicae Flos, thus providing a theoretical foundation for its cultivation and breeding.

Objective To investigate the clinical effects,feasibility,points for attention and complications of treating distal radial fractures through minimally invasive palmar locking plate.Methods A total of 22 patients with distal radial fractures managed by minimally invasive palmar locking plate from August 2009 to August 2010 were enrolled in the study.According to AO classification,there were five patients with type A2 fractures,three with type A3,four with type B1,seven with type B3,two with type C1 and one with type C2.Two vertical or parallel palmar incisions of 2 cm long were performed.The distal transverse incision was along the proximal wrist crease,while the distal and proximal vertical incisions were close to the radial side of flexor carpi radialis.Then,palmar locking plate was inserted through deep pronator quadratus.Results Follow-up period was 10-18 months(mean,12 months).According to Dienst function evaluation criteria,the results were excellent in 13 patients,good in six,fair in three and poor in zero,with excellence rate of 86％.Complications were as follows:one patient had delayed healing of proximal wrist crease incision;one patient had numbness of thenar eminence and the symptom was disappeared three months postoperatively;two patients remained pain on ulnar side of the wrist joint;two patients had limited function of the wrist joint.Conclusions Minimally invasive palmar locking plate is safe,mini-invasive and reliable when treating distal radial fractures,which is conducive to early functional exercise.Furthermore,the method achieves satisfactory appearance of the incision which meets the minimal invasion and thus deserves clinical practice.

Objective:To explore the clinical diagnosis and treatment methods for nasal septum malignancies. Methods:A retrospective study of seven cases of primary nasal septum malignancy was conducted. These cases were treated at the Tianjin First Central Hospital between February 1993 and October 2009. Results:Among the seven cases, one was reported malignant lymphoma, which underwent chemotherapy and radiation therapy;the other six malignant tumors included three cases of adenoid cystic carcinoma, one malignant melanoma, one chondrosarcoma, and one papilloma. Patients were treated by nasal endoscopy, external approach, lip midline incision, overturned facial surgery, chemotherapy, and radiation. During follow-up, which lasted for 2 years to 16 years from time of surgery, two fatalities resulting from recurrence or metastasis were recorded. One of these fatalities occurred after 2 years, and the other was recorded at 3 years and 4 months after surgery. Four patients survived without recurrence. Conclusion:The key treatment for nasal septum malignancies was one-time radical resection. Combined postoperative treatment and follow-up should be conducted.

The combined use of chemometrics and chemiluminescence（CL）measurements,with the aid of the stopped-flow mixing technique,developed a simple time-resolved CL method for the simultaneous determination of captopril（CPL）and hydrochlorothiazide（HCT）.The stopped-flow technique in a continuous-flow system was employed in this work in order to emphasize the kinetic differences between the two analytes in cerium（IV）-rhodamine 6G CL system.After the flow was stopped,an initial rise of CL signal was observed for HCT standards,while a direct decay of CL signal was obtained for CPL standards.The mixed CL signal was monitored and recorded on the whole process of continuous-flow/stopped-flow,and the obtained data were processed by the chemometric approach of artificial neural network.The relative prediction error（RPE）of CPL and HCT was 5.9% and 8.7%,respectively.The recoveries of CPL and HCT in tablets were found to fall in the range between 95% and 106%.The proposed method was successfully applied to the simultaneous determination of CPL and HCT in a compound pharmaceutical formulation.

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

Objective To investigate the influencing factors and technical points of induced membrane technique for treatment of bone defect.Methods All of 20 patients of bone defect were treated by induced membrane technique in our hospital from January 2008 to November 2014,including 15 males and 5 females;aged 13-69,average 38.5;infectious bone defect in 16 cases and non-infectious bone defect in 4 cases.Record the complications,evaluate the healing of bone defect and functional recovery of adjacent joints by Paley method,respectively,and grade the range of movement (ROM) of adjacent joints by authors's method.Results In the first stage of surgery,1 case needed a second operation as the wound gradually spitted and bone cement contaminated after tighten closure of the skin flap,while others had no infection or recurrence of infection.In the second stage of surgery,3 cases had induced membrane damage and defect.All were followed-up from 12 to 50 months (average 19.7 months);all the bone defects healed,the clinical healing time was 3.0 to 7.0 months (average 4.7 months).The healing time in the 3 cases with induced membrane damage and defect (average 6.0 months) was longer than that in patients without induced membrane damage and defect(average 4.6 months).1 case of infectious bone defect with induced membrane damage and defect had local infection in 6 months after the second stage of surgery,for whom the conservative treatment was invalid but got controlled after second operation while 1 case of infectious bone defect without induced membrane damage and defect had local infection in 12 months after second stage of surgery,in whom the infection was controlled by the conservative treatment,the others had no infection or recurrence of infection,no broken of fixators noted;at the last follow-up,all the bone defect healing graded excellent,the functional recovery of the adjacent joints graded:excellent in 8 cases,good in 10 cases,and fair in 2 cases (the excellent and good rate was 90％),the ROM of the adjacent joints graded:excellent and good in 8 cases,respectively,fair and poor in 2 cases,respectively (the excellent and good rate was 80％).Conclusion Induced membrane technique has advantages of simple surgery,faster healing of bone defect,no correlation between the healing time and the length of bone defect,fewer complications,etc,but in clinical application,the operators must understand the therapy principle and pay attention to the influencing factors and technical points so as to avoid operation errors,reduce complications and improve therapeutic effect.

Objective To discuss the surgical techniques and results of internal fixation supplemented with soft tissue repair of triad injury of the elbow via the combined anterior-lateral approach.Methods A retrospective analysis was done on 15 patients with triad injury of the elbow treated from January 2011 to August 2014.There were 10 males and 5 females,aged 38.4 years (range,20-61 years).Injury resulted from traffic accidents in nine patients,high-level falls in three and groundlevel falls in three.Radial head fractures were Mason type Ⅰ in five patients,type Ⅱ in eight,and type Ⅲ in two.Fractures of the coronoid process of the ulna were Regan-Morrey type Ⅰ in four patients,type Ⅱ in ten and type Ⅲ in one.Time interval between injury and operation was 8.5 d.All patients were firstly operated on through the anterior approach to have Herbert screw fixation or anterior capsule suture lasso fixation of the coronoid fracture as well as Herbert screw fixation of the radial head fracture.And then,elbow lateral incision was made to repair lateral collateral ligament complex and common extensor tendon by 2-0 Ethibon suture.Elbow flexion-extension,forearm pronation-supination and Mayo elbow performance score (MEPS) were used to evaluate the clinical elbow functions.Complications were recorded after operation.Results All patients were followed up for mean 19.6 months (range,12-30 months).All coronoid fractures healed.Radial head fractures in 14 patients healed.Comparison of preoperative to final follow-up variables presented significant differences in elbow flexion-extension [(45.1 ± 5.6) °:(129.5 ± 9.3) °],forearm pronation-supination [(55.4 ±.8.7) °:(140.5 ± 10.3) °] and MEPS [(25.1 ± 9.6) points:(91.2 ± 5.2) points] (P ＜ 0.01).Complications included heterotopic ossification in one patient and nonunion of radial head fracture in one patient.No neurovascular injury of the elbow,elbow residual instability,dislocation,elbow stiffness and active pain occurred.Conclusion Combined anterior-lateral approach with internal fixation and soft tissue repair is a simple and safe method that effectively restore the elbow joint function.

OBJECTIVE Toexplorethefeasibilityofpreparingamigrainemodelusingratdura materofsuperiorsagittalsinusinfollowingelectricalstimulation.METHODS MaleSDratswereexposed to 1 h electrical stimulation after brain electrode implantation,at 3 V,6 Hz,and a pulse of 0.25 ms.At 0, 1 5,30 min,and 1 h,orbital blood and brain tissue were taken.The content of calcitonin gene-related peptide (CGRP),substance P and prostaglandin E2 (PGE2 )in the plas ma and brain tissue was detec-tedbyradioimmunoassay.RESULTS ThebraintissuecontentofCGRP,substancePandPGE2in the sham group and normal group was not significantly different.CGRP,substance P and PGE2 in brain tissue of model group were 3.41 ±0.93,1 .80 ±0.64,3.41 ±0.93 and (1 .80 ±0.64)ng·g -1 respec-tively,significantly different from sham group(P<0.05).At different time points,the content of CGRP, substance P and PGE2 in plasma orbital venous plexus showed no significant difference between sham groupandmodelgroup.CONCLUSION ThereleaseofCGRP,PGE2andsubstancePinbraintissue is significantly different fro m that of plas ma after electrical sti mulation.The results suggest that a model of migraine can be constructed by electrical stimulation.