Objective To investigate the expression of neurotrophin- 3 (NT- 3) gene in Schwann cells (SCs) of rat sciatic nerve introduced by adenovir al vector in vivo. Methods Recombinant adenoviral vector for NT- 3 was propagat ed in 293 packaging cells and was titered by tissue culture infections dose 50 ( TCID50) method. Ad- NT- 3 was injected intraneurally into the rat sciatic nerv e following axotomy of the sciatic nerve which was repaired after the immediate injury. Immunohistochemical staining was employed to determine the expression of NT- 3 in SCs in rat sciatic nerve, and the expressive intensity difference was measured with LEICA M550 imagine analysis system on the tissue slides. Results Two days after injection of NT- 3 into the rat nerve, positive stain in the SCs were apparent in the vicinity of anastomosis. There was significant increase of the amount of NT- 3 expression 7 days following injection of Ad- NT- 3 (vs. 2 days group P0.05). Compared with the 2 days group, the 14 and 28 days groups still maintained a relatively high leve l of NT- 3 (P

Objective The articular neurohistology is a new approach to explore stabilized and chronic pain mechanisms for joint research. Furthermore, the distribution of mechanoreceptors and their functional condition play an important role in the movement and stability of joint. The animal experimental study suggest that there is a various characteristics of the type and amount of mechanoreceptors in different joints. Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs as joint mechanoreceptors are widely distributed on nearly all of human joint, and have impact on stabilization of joint. The present investigates the neurohistological features of Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs in the rotator cuff and the capsule, and explores their behaviors for maintaining stability of the shoulder joint. Methods Six rotator cuffs and capsules of shoulder joint were obtained from cadavers died of the accident. The tendinous tissue of subscapularis, supraspinatus、infraspinatus, and teres minis were resected from the interface of tendon and muscle to great and lesser tuberosity of humerus, the capsule tissues of glenohumeral were harvested as well. Then, the tissues were cut into a small fragments fixed with the formalin buffer solution, and embed in paraffin. The section was made along the longitudinal axis of the tendon at 150 ?m interval with Leitz 1516 machine. The modified Bielschowsky staining and the S-100 immuohistochemical staining were adopted to observe the mechanoreceptors of Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs. The distribution density and size of the sensitive nerve endings were analyzed with Leica imaging process system. Results The tendinous part of rotator cuff could be divided into three layers: synovial, intermediate and articular layer. The morphology of the these three types of mechanoreceptors, Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs, could be identified clearly in the every layer of the tendious tissues under microscopic examination. Pacini corpuscle was ellipse and had multi layer capsule with the thickness of 20-40 ?m; Ruffini corpuscle was columnar and 350-550 ?m in size; Most of Golgi tendon organs were found at the interface of tendon and muscle. Among of them, the Pacini corpuscle and Ruffini corpuscle were much more than the Golgi tendon organs in rotator cuff and capsule of shoulder. The results of immuohistochemical staining and the counting of nerve ending assisted by the computer showed the number of sensory nerve endings in the subscapularis, supraspinatus and anterior capsule of the shoulder were much more than in the infraspinatus, teres minis and posterior capsule. The diameter of nerve bundle in the subscapularis, supraspinatus and anterior capsule was thicker than in the other periarticular structures. In periarticular structures, the mean value of nerve fiber counting were 25 in infraspinatus, 25 in supraspinatus, 13 in subscapularis, 10 in teres minis, 22 in anterior capsule and 12 in posterior capsule. Conclusion Shoulder capsule and rotator cuff are rich in the three type of mechanoreceptors, Pacini corpuscle, Ruffini corpuscle and Golgi tendon organs. However, there are a different distribution density and size of them in rotator cuff and capsule of shoulder, but the three of mechanoreceptors are mainly concentrated in the subscapularis, supraspinatus and the anterior capsule. S-100 immuohistochemical staining is much more sensitive than the modified Bielschowsky silver staining in displaying the innervations of the rotator cuff and capsule. The characters of the sensory nerve ending distribution in shoulder imply that the innervations of the shoulder play an important role in keeping the stability and mobility of the shoulder.

[Objective]To investigate the morphological changes of the roof of the subacromial bursa and the extent of involvement of the subacromial bursa after rotator cuff tear.[Method]For the experimental group,the roof of the subacromial bursa was obtained from patients with rotator cuff tear both at the tear site and 2.5 to 3.0 cm distal to the tear site during surgery of rotator cuff repair.For the control group,the roof of the subacromial bursa was obtained during surgery of recurrent shoulder dislocation or humeral shaft fracture at the exposure site.The specimens were processed for histological staining,immunohistochemical analysis and for transmission electron microscopy.The number of cells were quantitated by counting cells with blue-stained nucleus in the subacromial bursa using a computer image analysis system.[Result]The number of cells increased significantly in the roof of the subacromial bursa obtained from both sites,the tear site and the distal site,in the patients with rotator cuff tear,compared with the normal bursa(P

[Objective]On May 12,2008,an over 8.0-magnitude earthquake occurred in Wenchuan county,Sichuan province,China.The anthor's field hospital was deployed to An county,which arrived on the 3rd day after the disaster.During the first week of the deployment,284 victims with musculoskeletal injuries received orthopedic management.The objective of this article is to highlight the experience gained by this author's medical assistance team,focusing on the orthopedic care in the early part of the post-disaster relief activities.[Method]Records of 284 patients who underwent orthopedic management were reviewed retrospectively.[Result]There were a large number of musculoskeletal injuries amongst the survivors in the acute phase of the disaster,including spinal trauma,fractures and dislocations,peripheral nerve injuries and soft tissue injuries.The average age was 49.3 years(range,11 to 92 years) with 133 men and 151 women.The patients with upper extremity fracture were 85,with lower extremity fracture 73,with spinal fracture 23,with shoulder or hip dislocation 7.The patients with hand or foot injuries,peripheral nerve injuries and soft tissue injuries were 96.Open injuries were 67,the remaining were closed injuries.Totally 131 patients recovered,and the rest were transferred to the rear hospital for further management.The X-ray postoperatively showed that 21 open fractures were reduced functionally by means of external fixator,and the shoulder or hip dislocations were reduced anatomically.The wounds with primary suture had no infection.There were no other complications.[Conclusion]Correct diagnsis of the injuries,providing scientific and resonable therapeutic measures,and correct operative indication should be emphasized for early orllopedic care.Definitive surgeries such as open reduction and internal fixation are not recommended strongly.External fixation for post-earthquake open fractures is simple and effective,allowing simple approach to wound care and transferring to rear hospital during the whole period of fixation.

AIM:To develop a HPLC method for determining scutellarin and geniposide in Yinshanlian Granules(Herba Artemisiae scopariae,Herba Scutellariae barbatae, Fructus Gardemiae,etc). METHODS: The analysis was performed on Diamonsil C_(18)(200 mm?4.6 mm,5 ?m) with mixture of acetonitrile(A) and 0.1% phosphoric acid solution as mobile phase in gradient mode.The concentration of solvent A were 5%,33%,5% and 5% at 0,30,31 and 35 min,respectively.The detection wavelength was set at 240 nm and the column temperature was at 30 ℃.(RESULTS:)The linear calibration curves were obtained in the concentration range of 0.032-0.288 mg/mL for scutellarin and 0.009 6-0.086 4 mg/mL for geniposide.The average recoveries of scutellarin and geniposide were 100.6%(RSD=0.80%,n=9),102.6%(RSD=1.1%,n=9),respectively.CONCLUSION: The method is quick,simple,and reproducible,which can be used to control the quality of Yinshanlian Granules.

Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.

Objective:To investigate the diagnostic value of different detection methods for Mycobacterium tuberculosis in bronchoalveolar lavage fluid (BALF) from patients with pulmonary tuberculosis.Methods:BALF from l00 patients in Changsha Central Hospital from January 2013 to December 2015 was collected.Among 100 patients,65 cases were clinically diagnosed as tuberculosis,and 35 cases served as control.BALF smear method,polymerase chain reaction (PCR) and membrane reverse dot blot (RDB) were used for synchronous detection of Mycobacterium tuberculosis.Results:The positive rates by BALF smear method,PCR and RDB were 43.08％,73.84％ and 92.31％,respectively (P＜0.05).Sensitivity,specificity,accuracy,and negative predictive value for BALF smear were 43.08％,88.57％,59.00％,and 45.59％,respectively;for PCR were 73.85％,100％,83.00％,and 67.31％,respectively;for RDB were 92.31％,100.00％,95.00％,and 87.50％,respectively.Conclusion:The technique of membrane RDB can not only accurately diagnose Mycobacterium tuberculosis,but also can rapidly and easily identify the resistance of Mycobacterium tuberculosis to streptomycin (SM),rifampicin (RFP) and isoniazid (INH) genotypes.It possesses high clinical value.

0bjective To establish a standard curve method with more accuracy employed in fluorescence real-time PCR(RT-PCR)as a alternation of the general method.Methods β-actin and KLK11 plasmid DNA for quantitative standard curve were constructed in our study,and Plasmids of β-actin was employed as a internal control.After serial dilution these plasmid were used as DNA standard to obtained slope.Expression of these two genes in malignant prostate cancer cell line LNCAP were tested by real-time PCR,and we analyzed the RT-PCR results with two different methods and compared their accuracy.Results Thestandards curves made from these linear DNA standards showed good linearity (R2=0.991 and 0.992 for β-actin and KLK11 standards graphs),but also displayed a discrepancy in their PCR efficiency(β-actin 123% and KLK11 99%).There were different results after two different stand curve analytical method:the expression of KLK11 mRNA in LNCAP was downregulated in general standard curve method.In the new analytical method,howerer,KLK11 upregnlated for 4.46-fold.And there was a significant difference between aplification efficiency of targt gene and internal control gene(t=4.829,P<0.05).Conclusions Compared with general standard curve method,the new advanced standard curve method described here avoids an error which considers there is identical amplification efficiency between target gene and internal reference gene.It is considered to be a more correct analytical method in fluorescence real-time PCR.

Objective To investigate the ultrasound characteristics of pseudothrombophlebitis.Methods The ultrasound characteristics of thirty-two popliteal cyst cases with pseudothrombophlebitis which were confirmed by MRI or puncture were retrospectively analyzed.The former group was matched with a case control group of sixty-four patients with asymptomatic popliteal cysts,the ultrasound images were comparative analysis between the two groups.Results Compared to the control group,both the length and width of the case group were larger [(12.4 ± 4.7) ×(2.5±0.4)cm vs (5.3±2.9) × (1.2±0.4)cm,P ＜ 0.001],and there were 18(43.8％) cases with the cysts extension into the calf.The case group were more prone to show cyst with poor ultrasound penetration or solid-cystic echo (78.1％ vs 9.4％,P =0.000),which were diagnosed as popliteal cyst with hematoma or infection.Six cases of case group showed irregular anechoic area surrounding the inferior border of the cysts,which were diagnosed as ruptured popliteal cyst,no similar imaging detected in the control group(P =0.001).Both the two group showed septation in the cyst,but it did not differ significantly(P ＞0.05).Conclusions Ultrasonography is helpful to the diagnosis of pseudothrombophlebitis,the ultrasound characteristics include large cyst extension into the calf,cyst with poor ultrasound penetration or solid-cystic echo and irregular anechoic area surrounding the inferior border of the cysts.

Purpose To study the expression of TMPRSS2, ERG and ETV1 in prostatic cancer and their clinical pathologic signifi-cance. Methods Tissue microarray and immunohistochemistry (MaxVision) were used to detect TMPRSS2, ERG and ETV1 expres-sion in 70 prostatic cancer tissues, 10 prostatic intraepithelial neoplasia tissues and 18 benign prostate tissues. Results There was no statistical significance on positive rate of the expression of TMPRSS2 among prostatic cancer tissues, prostatic intraepithelial neoplasia tissues and benign prostate tissues (P>0. 05). The positive rate (81. 4%) of ERG in prostatic cancer tissues was significantly higher than that in prostatic intraepithelial neoplasia tissues ( 30. 0%) and benign prostate tissues ( 0. 0 ) ( P <0. 05 ) . The positive rate (68. 6%) of ETV1 in prostatic cancer tissues was significantly higher than that in prostatic intraepithelial neoplasia tissues (50. 0%) and benign prostate tissues (22. 2%) (P<0. 05). There was no correlation among the positive rates of TMPRSS2, ERG and ETV1 in prostatic cancer tissues and age (P>0. 05). The expression of TMPRSS2, ERG and ETV1 was positively correlated to Gleason score and clinical stage (P<0. 05). TMPRSS2 had positive correlation with ERG (rs =0. 465, P<0. 01). TMPRSS2 had positive correla-tion with ETV1 (rs =0. 590, P<0. 01). ERG had no positive correlation with ETV1 (rs =0. 151, P>0. 05). Conclusion ERG and ETV1 are expected to become therapeutic targets for prostate cancer. Detecting TMPRSS2, ERG and ETV1 at the same time is helpful to diagnosis and differential diagnosis of prostatic cancer, which might be new molecule markers of prostate cancer.