Objective To investigate the anti-tumor effects of combined recombinant murine interleukin-12 and GM-CSF gene adenovirus vector(AdCMVIL-12,AdCMVGMCSF) on B16 melanoma.Methods B16 melanoma cells were injected subcutaneous at right back of C57/BL6 mice,48 tumor bearing mice were divided into 4 groups at random,when tumor diameter attained to 5 mm,AdCMVIL-12,AdCMVGM-CSF and AdCMVIL-12+AdCMVGM-CSF and AdCMVLacZ(control) were injected in different groups,tumor growth and immune responses of each group were tested,and the IL-12 and GMCSF levels in serum were observed.Results In combined group,necrosis, lymphocytes and macrophages were found in tumor tissues.30 d after treatment,the average volume in combined group was(60.73?11.36)mm~3, compared with AdCMVLacZ(control) group,and AdCMVIL-12 group,AdCMVGM-CSF group [(675.18?80.59),(186.91?19.15),(223.45?23.11)mm~3],there were significant differences(P

Objective To observe the effect of tacrolimus(FK506)in promoting repair of the injured spinal cord pathway after neural stem cell transplantation in rats. Methods A neurysm clip was used to compress the T8 spinal cord segment of SD rats under microscope to establish model of spinal cord injury.The rats were randomly divided into three groups seven days after injury,ie,control group (injection of normal saline at the injury center),transplantation group(injection of neural stem cells,NSCs,at the injury center),FK506 group(injection of NSCs at the injury center plus 7 days of intrapernerve conduction was compared by using the Basso-Beatfle-Bresnahan (BBB) scale,BDA tracing,somatosensory evoked potential(SEP)and motor evoked potentials(MEP)monitoring at 1,2,4 and 8weeks. Results The motor function of the hind limb after injury was recovered in various degrees with time,with the most significant recovery at 4 weeks.The BBB score reached 6,the maximum at 8 weeks.BDA tracing showed that some nerve fibers were found crossing the injured center of the spinal cord one week later in FK506 group and cell transplantation group,that BDA-positive labeled corticospinal tract fibets were seen across the injury site in all groups by the end ofthe eight weeks.In the FKS06 group,the regeneration could be observed even as 1.7 cm away from tlle injury center.SEP latency was significantly shorter in the FK506 group after two weeks(P<0.05)and the MEP latency in the FK506 group was shortened significantly at four weeks compared with the other groups(P<0.05),indicating that the immunosuppressants could promote the recovery of the injured spihal cord,shorten the latency of SEP and MEP，improve SEP at early stage and MEP at late stage．Conclusions Systemic application immuno suppressive agents FK506 plays an important role in neuroprotection and neurotrophy，which promotes the repair of the injured spinal cord after neural stem cell transplantation．

Objective To investigate the effect of treating tibial plateau fractures involving the posterior column via anterolateral approach.To discuss its merit and demerit,indications and contraindications.Methods From Jan 2012 to Jan 2015,37 patients with closed tibial plateau fractures involving the posterior column were treated by 3.5mm proximal tibial plate with narrow plate head through standard anterolateral approach.The group included 25 males and 12 females,aged from 23 to 65 years old (average 44.6 years old).During the treatment and follow-up period,the curative effect was evaluated by using Rasmussen's radiological grading for early radiological outcomes and HSS grading,Lysholm grading,Lachman test and Pivot-shift testfor clinical examination at the 1 year follow-up.Results The average time for the operation was 98 min (range,55-170 min).1 year follow-up was completed in 37 patients.The average fracture union time and full weight bearing were 10.6 weeks (range,8-16 weeks) and 11.2 weeks (range,8-20 weeks) respectively without reduction loss.No statistical difference was found in either the tibial plateau angle (TPA) or posterior slope angle (PA) when comparing the results at instant,3rd month,6th month and 12 month.The mean score of the Rasmussen' s radiological grading was 15.6(range,12-18) and the mean score of the HSS grading was 88.6 (range,80-100).The average range of motion of the knee joint was 128.6°(range,110°-150°).The mean score of the Lysholm grading was 91.6±3.9. The Lachman test and the Pivot-shift test were negtive.Conclusion Treating tibial plateau fractures involving the posterior column by 3.5 mm proximal tibial plate with narrow plate head through standard anterolateral approach is an effective method.The protocol is simple and safe,the approach is familiar by most clinicians.Good reduction and fixation,earlier functional exercise can be achieved easily.The knee function recovered well and earlier curative effect was satisfied.

Objective To investigate the value of external fixator in the treatment of posttraumatic elbow stiffness.Methods From January 2007 to December 2009,25 patients with posttraumatic elbow stiffness received surgical arthrolysis with assistance of external fixator.There were 10 patients with grade 1 stiffness,12 patients with grade 2 stiffness and three patients with grade 3 stiffness.The range of motion (ROM) was 0°-75° ( average 33.4°) preoperative,with ＜ 30° in 12 patients.For the patients with grade 1 stiffness,intraoperative distraction with an external fixator rather than open arthrolysis was performed ; for the patients with grade 2 stiffness,the heterotopic ossification was removed through a limited approach before the external fixator was applied; for the patients with grade 3 stiffness,additional osteotomy and internal fixation before arthrolysis were performed.After operation,there was a rehabilitation phase,followed by mobilization of the elbow joint under protection of the external fixator for a mean period of six weeks.Results The intraoperative mean ROM was 105.6°,which was significantly improved than that before operation (P ＜0.05).The maximal flexion degree was above 110°in all patients.No instability occurred.Two patients were excluded from the follow-up:one had ulna fracture two weeks after operation due to an incorrect placement of the pin,and additional internal fixation was then carried out;another patient had numbness in the ulna side when flexed the elbow after arthrolysis,and the ulna nerve transposition was performed through another operation.Other patients were followed up for a mean period of six months.At the latest follow-up,the mean ROM was 97.4°( range,70°-130°),significantly improved than that before operation (P ＜ 0.05 ),and lest 8.2°on average (0°-15 °) compared with the intraoperative ROM (P ＞ O.05).The maximal flexion in all patients was above 100° Conclusion With the advantages of minimal invasive or noninvasive,indirect capsular traction,stability maintenance during the movement,and prevention of the motion loss during the rehabilitation,arthrolysis of the elbow joint with the use of external fixation frame can be a good alternative in the treatment of posttraumatic elbow stiffness.

Objective To investigate the injury mechanism, fracture characteristics, treatment strategy and prognosis of Pilon fracture without fibula fracture. Methods 28 cases of Pilon fracture without fibula fracture (29 fractures) treated in our hos?pital were adapted from January 2009 to December 2014. 22 cases were male, 6 were female, aged 46 years old (23-75). Cause of injury:tumble in 14 cases, fall from high place in 8 cases, traffic accident in 5 cases, bruise in 1 case. 10 cases had associated in?juries, and 2 cases were open injuries. The OA/ATO fracture type, Rüedi&Allgower fracture type, and characteristics of fractures were summarized. The appropriate treatments were selected, and the operative method, operative time, surgical approach, fixation, fracture healing time, ankle Mazur score, Burwell?Charnley reduction score and postoperative complications were recorded. Re?sults On the mechanism of injury, 28 cases mostly had a median?low energy injury mechanism with little open injuries. On the fracture characteristics, there was a 96.6%anterior distal tibia articular fracture involved with no other specific fracture character?istics occurred. The treatment of the fracture mostly received a single operative approach to complete open reduction and internal fixation operation. The average operation time was 119 minutes, the average fracture healing time was 7.1 months, the excellent and good rate of ankle Mazur score was 86.2%and the satisfactory rate of Burwell?Charnley fracture reduction score was 93.1%. Skin infections occurred in 3 patients after operation. After effective antibiotic and correct clinical treatment, the incisions got healed in these 3 patients. Conclusion Pilon fracture with intact fibula mostly is caused by median to low energy injury when an?kle at neutral or varus position, most with anterior distal tibia articular fracture by foot dorsal flexion. Generally use a single anteri?or approach can complete open reduction and internal fixation operation with satisfactory clinical treatment. This kind of fracture could achieve good reduction, good bone healing, good function and little complications after correct treatment.

Objective To investigate the morphological properties of posterior Monteggia lesion with associated elbow dislocation,to propose its injury mechanism,and to present its surgical methods and its outcomes.Methods The injury mechanism,radiographs and surgical records of patients with posterior Monteggia fractures and associated elbow dislocation were retrospectively reviewed from January 2011 to December 2013.11 patients were included,with 43.3 years old on average.10 were resulted from high-energy injuries.According to the Jupiter classification,Ⅱ A fracture-dislocation 9 cases,Ⅱ B fracture-dislocation 1 case and Ⅱ C fracture-dislocation 1 case.The general medical data,morphological properties,and the surgical methods.Functional outcomes were followed up.Results These cases had several intrinsic morphologic features:fractures of the coronoid tip and its anteromedial aspect;dislocation of the olecranon from the trochlear notch;fracture of the radial head and disruption of the lateral collateral ligament;a normal proximal radioulnar joint.Operation was carried out with emphasis on elbow stability restoration.All patients underwent the primary operations in the supine position and a routine posterior approach was used.An additional anterior approach was used in 1 case (Ⅱ B).The coronoid tip was first fixed,followed by the anteromedial coronoid fragment,the radial head,the olecranon,and then the LCL.An anatomic olecranon plate was used as the fundamental fixation device.Anteromedial coronoid fractures were mostly fixed with a T-shaped metacarpal plate.The coronoid tip fractures were fixed with screw,Kwire or suture,and the injured LCL was repaired with suture anchors.Ten of the 11 patients were followed up with a mean period of 22.7 months.The extension-flexion motion ranged from 45°-140° (104° average),and the pronation-supination motion ranged from 45° 180° (128° average).According to the Broberg and Morrey functional index,the excellent to good ratio was 70％.Conclusion Patients with posterior Monteggia fracture and associated ulnohumeral dislocation have small and comminuted coronoid fractures,and mostly have a normal PRUJ relationship.It might experience a different mechanism to those of the posterior Monteggia fracture without elbow dislocation.Emphases should be address to restore the rotation stability of the elbow when operation.

Objective : To investigate the epidemiological and clinical characteristics of cases with pertussis in Yiwu City, Zhejiang Province, so as to provide insights into pertussis diagnosis and control. Methods: A total of 186 patients with definitive diagnosis of pertussis in medical institutions in Yiwu City from November 1, 2020 to August 31, 2022 were recruited, and subjects' demographic, clinical symptoms and history of pertussis vaccination were collected using questionnaire surveys. The temporal, population, and spatial distribution and clinical symptoms of pertussis were analyzed using a descriptive epidemiological method, and the clinical characteristics of pertussis patients with different doses of pertussis vaccination were compared. Results: Pertussis was found to predominantly occur during the period between July and November (101 cases, 54.30%), and the three highest-incidence regions included Jiangdong Street, Beiyuan Street and Choujiang Street (87 cases, 46.77%). The 186 pertussis cases included 105 males (56.45%) and 81 females (43.55%), and included 144 cases with age of onset under 7 year (77.42%). Preschool and diaspora children were predominant among all pertussis cases, and the main clinical symptoms included spasmodic cough (97 cases, 52.15%), post-tussive vomiting (82 cases, 44.09%) and aggravated cough at night (77 cases, 41.40%). Routine blood tests measured 119 cases with abnormal white blood cell counts (63.98%), 137 cases with abnormal lymphocyte counts (73.66%), 39 cases with abnormal neutrophil counts (20.97%), 21 cases with abnormal platelet counts (11.29%) and 111 cases with abnormal hemoglobin concentrations (59.68%). There were 55 cases that were unvaccinated (29.57%), 23 cases that were not fully vaccinated (12.37%), and 108 cases that were fully vaccinated (58.06%). There were significant differences among pertussis cases with different doses of vaccination in terms of age, incidence of post-tussive vomiting, percentage of abnormal platelet counts and percentage of hemoglobin concentrations (all P<0.05). Conclusions The majority of pertussis cases are preschool and diaspora children in Yiwu City from November 1, 2020 to August 31, 2022, and the clinical symptoms mainly include spasmodic cough, post-tussive vomiting and aggravated cough at night, with atypical symptoms. The capability for differential diagnosis of pertussis is required to be improved in medical institutions.

Objective: To explore the revision strategy of the malunited tibial plateau fracture and to analysis the main points of four common revision operations and the clinical effect. Methods: From January 2012 to December 2016, 18 patients (5 males and 13 females) aged 35-60 years (average 49.7 years) underwent tibial plateau revision surgery in our hospital were collected. The time from the second revision operation to the first operation was 2-24 months (average 10.4 months). Our revision strategies were as follows: Firstly, to determine whether there is an infection or not. If there was infection, we changed the original internal fixation to external fixator to control infection. Secondly, todetermine whether the patient could suffer re-reduction and internal fixation. If the patient was older (>65 years old) or with severe local bone defect, total knee arthroplasty should be performed. Thirdly, patients were divided into four operation modes according to the tibial plateau fracture malunion type: 1. the original fracture line osteotomy; 2. the tibial tubercle + original fracture line osteotomy; 3. tibial metaphyseal window-rod reduction; 4. the osteotomy of fibula head and original fracture line osteotomy. Results: All patients were followed up for 12-30 months (average 16.8 months), and the operation time was 120-300 min (average 185 min). 2 cases were infected before operation and the original internal fixation were removed to instead of external fixator;1 patient underwent total knee arthroplasty; 3 cases were treated with metaphyseal open window-rod reduction and internal fixation; 6 cases were operated with the original fracture line osteotomy and internal fixation; 4 cases were treated with tibial tubercle osteotomy+original fracture line osteotomy and internal fixation; 2 patients underwent fibular head osteotomy+ original fracture line osteotomy and internal fixation. All patients achieved bony union at the last follow-up. The healing time was 3-6 months (mean 3.6 months). The postoperative knee Rasmussen score was 19-29 (average 22.9), compared with average 14.4 points before operation (t=-10.169, P=0.001). The postoperative range of motion of knee joint was 60-110 degrees (mean 94.5 degrees), compared with average 55 degrees before operation (t=-5.773, P=0.001). The post-operative VAS pain score was average 1.1 points, compared with average 4.2 points before operation (t=8.960, P=0.001). Fracture reduction was excellent in 12 cases and good in 5 cases, with the excellent and good rate of 100%. 3 patients still had 2mm collapse on the articular surface, while 3 patients still had mild valgus (less than 5 degrees). There were 2 cases of superficial infection of the wound surface after operation. Conclusion It was difficult to revise the malunion of tibial plateau fracture and it was necessary to make a detailed operation plan before the operation. Satisfactory clinical effects could be obtained for the patients by correct revision strategy. The key to success was the proper revision strategy which was adopted according to the different characteristics of the tibial plateau fracture malunion of the patients.

BACKGROUNDTo express and purify human alpha3-integrin to serve as the antigen to prepare its antibody and to separate the Vero cell clones without expression of alpha3-integrin.

METHODSThe human alpha3-integrin gene was amplified by using RT-PCR, then subcloned into a pQE30 expression vector and expressed in E. coli. The gene expression was confirmed by Western blot assay. Rabbit was inoculated with purified antigen to stimulate the antibody generation. The target Vero cells were separated by negative selection using antibody plus complement mediated cytolysis. The separated cell clones were confirmed by immunofluorescence and Western blot assay.

RESULTSThe alpha3- integrin gene was cloned and expressed effetively, Western blot assay revealed that the expressed protein held good immune reactivity. High titer antibody was generated. However the expression of alpha3-integrin was not detected on Vero, VeroE6, Hep-2, 2BS and 293 cells.

CONCLUSIONSThe results of the study suggested that hantavirus has other receptors on Vero cells beside alpha 3-integrin.

Animals ; Cercopithecus aethiops ; Cloning, Molecular ; Gene Expression ; Hantavirus ; Integrin beta3 ; biosynthesis ; genetics ; immunology ; Rabbits ; Receptors, Virus ; Vero Cells ; metabolism