Objective To find an ideal method of DNA isolation from blood and especially from clotted blood and to minimize the volume of blood collected for laboratory and clinical tests.Methods DNAs were isolated from antiagglutinated and agglutinated blood samples from auricular veins of 30 healthy subjects. The DNAs of these samples were obtained by a nonenzymatic, nontoxic procedure optimized by us and determinated by agarose gel electrophoesis and PCR. Results The yields of DNA isolated from clotted blood and antiagglutinated blood were (40.2?8.86)mg DNA/L and (39.1?10.2)mg DNA/L, and purities were 1.87?0.11 and 1.92? 0.12. The DNAs that we isolated from all samples had high molecular weight and by PCR the dimorphism of ALU alleles of the 8th intron of t-PA was easy to be obtained, so they were complete and reliable. Conclusion This method is rapid, easy, efficient and nontoxic for isolation of DNA from clotted and fresh blood and meets requirements for clinical testing and molecular biology study.

Objective To optimize the extraction process for the effective fraction of Bushen Yizhi ( BSYZ) formula. Methods Yield of the effective fraction and contents of two marker compounds 2,3,5,4 -tetrahydroxystilbene-2-O-β-D-glucoside and osthole assayed by high performance liquid chromatography (HPLC) were used as the optimizing indexes, and weighted coefficient of the indexes was analyzed by analytic hierarchy process ( AHP). Ratio of liquid-to-solid, extraction time, and extraction times were used to screen the optimal conditions by orthogonal test. Results The optimized extraction process of the effective fraction of BSYZ formula was as follows: extracting with 8-fold water for three times, and boiling for 1.5 hours each time. The verification test showed that the comprehensive scores of the optimized extractive process were higher than those obtained in the orthogonal test. Conclusion The optimized extraction process and conditions are practical in the manufacture of the effective fraction of BSYZ formula.

Objective To compare the position,volume and matching index (MI) of esophagus between quiet end-inspiration and end-expiration in three dimensional CT (3D-CT) assisted with active breathing control (ABC) and the corresponding phases in four dimensional CT (4D-CT).Methods Eleven patients with peripheral lung cancer underwent 4D-CT simulation scan and 3D-CT simulation scans in end-inspiratory hold (CTEIH) and end-expiratory hold (CTEEH) in succession.The 0％ phase was defined as end-inspiratory phase (CT0),while the 50％ phase was defined as end-expiratory phase (CT50).The proximal,mid-,and distal thoracic esophagus were delineated separately on CT0,CT50,CTEIH and CTEEH images.The position,volume and MI of each segment esophagus between CT0 and CTEIH,CT50 and CTEEH were compared.Results In the left-right (x) direction,the position differences in the proximal,mid-,and distal thoracic esophagus between CT0and CTEIH were (-0.02 ±0.16)cm,(0.06 ± 0.26)cm and (0.10 ± 0.33) cm respectively,and in the anterior-posterior (y) direction,the position differences were (0.04 ±0.24)cm,(0.04 ±0.12) cm and (0.08 ±0.15) cm respectively,and the position differences in the same direction were not statistically significant.In the x direction,the position differences of the proximal,mid-,or distal thoracic esophagus between CT50 and CTEEH were (-0.02 ±0.24) cm,(0.12 ± 0.37) cm and (0.26 ± 0.33) cm respectively,and in the y direction,the position differences were (0.03 ±0.21)cm,(0.04 ±0.17)cm and (0.14 ±0.18)cm respectively,and the position differences in x and y directions of proximal and mid-thoracic esophagus between CT50 and CTEEH were not statistically significant,while the position differences in x and y directions of distal thoracic esophagus between CT50and CTEEH were both statistically significant (t =0.025,0.024,P ＜ 0.05).The volumes of the proximal,mid-and distal thoracic esophagus were all larger in CT0and CT50 than those in CTEIHand CTEEH,but without statistical differences.The MIs of the volumes of the proximal,mid-and distal thoracic esophagus between CT0 and CTEIH were (0.50 ± 0.17),(0.50 ± 0.19) and (0.56 ± 0.08),respectively,and those between CT50and CTEEH were (0.50 ±0.16),(0.47 ±0.14) and (0.51 ±0.15),respectively.The MI of each segment esophagus between CT0and CTEIHwas larger than that between CT50 and CTEEH,but without statistical differences.Conclusions The influence of breathing modes on the centroid positions of the proximal,mid-thoracic normal esophagus were not significant and there were spatial mismatches for any segment esophagus between 3D-CT assisted with ABC and 4D-CT.

Objective To investigate the feasibility of amplitude of intraoperative nerve action potentials (NAP) for early quantitative diagnosis of peripheral nerve injury. Methods The sciatic nerve injury model were established in 16 rabbits. Intraoperative NAP were recorded after 4 weeks. According to amplitude of NAP, the injuried nerve were divided into 3 groups: NAP ＜ 100 μV in A group, 100 μV ≤NAP ＜ 500 μV in B group, NAP ≥ 500 μV in C group. Nerve specimen 1cm distal to injuried point were resected that received glycine silver stain and image analysis including number, diameter and cross section area of regenerative axons. Footprint parameter and ulcer area were measured and contrasted between each two groups. Results The number, diameter and cross section area of A group regenerative axons have significant difference with B and C group, no significant difference between B and C group; Footprint parameter and ulcer area have significant difference in each two groups. Conclusion Amplitude of intraoperative NAP can be a quantitative criteria to diagnose the degree of peripheral nerve injury that provides experiment evidence for guide intraoperative decision-making in clinical practice.

Objective To evaluate the effectiveness of using adipose-derived stromal cells (ADSCs)into a tissue-engineered peripheral nerve on bridging sciatic nerve gaps.Methods Forty-eight F344 female rats weighing 200 - 220 g were randomly divided into 6 groups of nerve grafting to repair 15 mm long asiatic nerve lesions,with 8 mrs in each group.Group A:ADSCs-laden acellular nerves;group B:differentiated ADSCs-laden acellular nerves;group C:Schwann cells-laden acellular nerves;group D:acellular nerves without cells;group E:autografts;group F:nonimplanted grafts.The effects were evaluated in terms of electrophysiulogy,Fluorogold retrograde tracing,histology and tracking studies.Results At 12 weeks after surgery,there was no graft bridging nerve gap in nonimplanted grafts.All the examinations of group A and B were better than group D,respectively (P＜0.05 or P＜0.01).But there were no statistically significant differences among group A,B,C,and D (P＞0.05).Conclusion ADSCs and differentiated ADSCs could promote nerve regeneration when used as seed cells to build tissue-engineered peripheral nerves with acellular nerve scaffolds.

AIM: To explore the neuroprotective action of progesterone(PROG), which has been proved to be a "neuroactive steroid". METHODS: The model of focal cerebral ischemia was established in rats by reverserble inserting a nylon thread with a diameter of 0.2 mm into the anterior cerebral artery through the internal carotid artery. The effect of PROG was assessed by determining water,sodium, potassium, and calcium contents in striatum of rats subjected to 2 h ischemia followed by 22 h reperfusion. RESULTS: The water,sodium,and calcium contents of middle cerebral artery occlusion(MCAO) striatum were obviously higher,the potassium content was obviously lower than those of non-MCAO striatum in I/R and DMSO groups,but there was no significant difference between these two groups.Compared with the result in I/R and DMSO groups , water, sodium and calcium contents significantly decreased, but potassium(P0.05). CONCLUSION: Treatment with PROG can significantly reduce the striatal injury of rats with cerebral ischmia-reperfusion.

Objective To investigate CT appearances of abdominal primary malignant fibrous histiocytoma(MFH).Methods The CT characteristics,clinical features and pathological data of 1 7 patients with MFH proved pathologically were analyzed retrospectively. Results The lesions located in retroperitoneum were 6,in liver were 5,in kidney were 2,in superior mesentery was 1,in greater omentum was 1,in stomach was 1,in ileum was 1.The lesions are oval shape,lobulated,nodule shape,and the size of these lesions were large. 2 cases of MFH located in gastrointestinal tract were slightly low density,and the remaining were uneven high density due to necro-sis.In CT contrast enhanced scan,the solid portion and internal divisions showed progressive or continuous enhancement,and the nec-rosis were not enhanced in MFH located in the retroperitoneum,the greater omentum,the superior mesentery and the liver.MFH in kidney was poorly circumscribed and showed mild progressive enhancement lower than normal renal parenchyma.The stomach and ileum lesions showed uniform and continuous enhancement with normal gastrointestinal mucosa in corresponding parts.Conclusion Imaging features of retroperitoneal MFH were the same as those of interstitial tumors,and most tumors showed features of progres-sive and persistent enhancement,but have different imaging appearances with the malignant lesions in corresponding parts.

Objective To analyze artifacts of gross tumor volume (GTV) and correlated factors in each phase images of four dimensional CT (4DCT) for peripheral lung cancer based on three dimensional CT (3DCT) assisted with active breathing control.Methods Nineteen patients with peripheral lung cancer underwent 3DCT (CTFB) and 4DCT simulation scans during free breathing and then underwent 3DCT simulation scans in end inspiration hold (CTEIH) and end expiration hold (CTEEH) assisted with active breathing control.The relative deviations (Devref) between the reference GTV (GTVref) and the GTVsdelineated based on CTFB (GTVFB) and all phases of 4DCT were calculated respectively.Correlations between GTVref and Devmax and between the tumor motion in the cranio-caudal (CC) direction and Devref were analyzed.Results The maximum median Devref of GTV was GTVFB with 17.83％,and the maximum median Devref of the GTV in all phases of 4DCT was GTV30 with 17.20％.A significant negative correlation was found between GTVEIH and Devmax (r =-0.691,P =0.001).The Devref was crrnelated with the tumor motion amplitude in the CC direction (r =0.323-0.617,P =0.005-0.150).The partial regression coefficient of influence of GTVref size and motion amplitude in the CC direction to the tumor Devmax were -0.500 and 0.583,P =0.002 and 0.001,respectively.Conclusions The GTV artifacts in different phase of 4DCT for the peripheral lung cancer were different to each other,and the influence of target displacement to artifacts was larger than that of target volume,so artifacts could be reduced by controlling breathing to reduce target displacement.

Objective To investigate the clinical efficacy of plate fixation with in situ reinforcing ulnar nerve and anterior transposition by olecranon osteotomy approach in the treatment of type C2 and C3 humeral intercondylar fractures.Methods A retrospective case control study was conducted to analyze the clinical data of 23 patients with humeral intercondylar fractures admitted to the Affiliated Hospital of Hangzhou Normal University from June 2014 to January 2017.There were 17 males and six females,aged 19-56 years,with an average of 41.2 years.According to AO classification,there were 15 patients with type C2 and eight with type C3.All patients were treated with plate internal fixation by olecranon osteotomy approach.The patients were divided into anterior group (11 patients) which was treated with ulnar nerve release anterior transposition and in situ reinforcing group (12 patients) which was treated with ulnar nerve release in situ.The operation time,intraoperative blood loss,fracture healing time,elbow flexion and extension range,elbow joint Mayo score,and excellent and good rate were compared between the two groups.Postoperative complications were recorded.Results All patients were followed up for 15-43 months,with an average of 26.7 months.All patients had no need of blood transfusion.There were no significant differences between anterior group and in situ reinforcing group in operation time [(219.1 15.1) minutes vs.(211.2 ± 17.1) minutes],intraoperative blood loss [(263.6 ± 35.3) ml vs.(237.5 ±25.6)ml] and fracture healing time [(18.2±2.4)weeksvs.(18.9±1.7)weeks] (P＞0.05).Nosignificant differences were found between the two groups in elbow flexion [(120.3 ±7.1) ° vs.(120.3 ±4.1)°],straightness [(3.7±2.7)° vs.(4.1 2.4)°],Mayo score [(89.09 ±9.17)points vs.(86.67 ±10.29)points] and excellent and good rate [91％ (10/12) vs.83％ (10/12)] (P ＞0.05).After operation,no complications such as infection,heterotopic ossification,ulnar neuritis,elbow valgus deformity,or internal fixation failure occurred.Conclusions For humeral intercondylar fracture,the plate fixation with in situ reinforcing ulnar nerve and anterior transposition by olecranon osteotomy approach both have advantages such as small surgical trauma,strong fixation,fewer complications,satisfactory recovery of elbow function.Ulnar nerve anterior transposition or not does not affect the occurrence of ulnar neuritis,and both can obtain satisfactory clinical efficacy.

Biopsy ; Endopeptidase K ; Fluorescent Antibody Technique ; Glomerulonephritis, IGA ; pathology ; Glomerulonephritis, Membranous ; pathology ; Humans ; Kidney ; pathology ; Lupus Nephritis ; pathology ; Paraffin Embedding ; Staining and Labeling