OBJECTIVE:To investigate the regulation effects of oleanolic acid on the mitochondrial nitric oxide synthase(mt-NOS) in human umbilical vein endothelial cells (HUVECs) with oxidative damage. METHODS:HUVECs in exponential phase were divided into normal group,model group and oleanolic acid low,medium and high dose groups(5,20 and 35 μmol/L). After drug acting for 24 h,all groups were given culture solution containing 100 μg/ml ox-LDL to reproduce oxidative damage except normal group. CCK-8 was used to detect cell viability. The mitochondria in cells were extrated,enzyme chemical method was used to detect mtNOS activity and mtNO content,fluorescence microplate method was used to detect fluorescence intensity of reactive oxygen species(ROS),and western blot was used to detect expression of cytochrome C(Cyto-C). RESULTS:Compared with nor-mal group,the cell viability in model group was decreased;mtNOS activity,mtNO content,ROS fluorescence intensity and Cy-to-C protein expression were increased,with significant differences (P<0.05). Compared with model group,the cell viability in oleanolic acid low,medium and high dose groups was increased;mtNOS activity,mtNO content,ROS fluorescence intensity and Cyto-C protein expression were decreased,with significant differences(P<0.05),and they had positive correlation with concentra-tions. CONCLUSIONS:Oleanolic acid can decrease the mtNOS activity of HUVECs,reduce the production of mtNO and Cyto-C, by a mechanism that may be related to the decrease of ROS expression.

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

BACKGROUND:A large number of studies have confirmed that tissue-engineered stem cel therapy is feasible to repair peripheral nerve injury, but the repair mechanism is unclear.
OBJECTIVE:To observe the differentiation and homing of bone marrow mesechnymal stem cel s under local nerve microenvironment by exploring the migration and effect of bone marrow mesenchymal stem cel s in the repair of damaged nerve.
METHODS:Male SD rats, aged 8 weeks, were selected to establish segmental nerve injury models by freezing the sciatic nerve. Thirty-six model rats were randomized into three groups (n=12):frozen nerve injury group, cel injection into the nerve group, cel injection around the nerve group. Before modeling and at 4, 8, 12 weeks after cel implantation, the sciatic nerve function index was measured. Electrophysiological test, contractility recovery rate, wet weight recovery rate of the triceps surae were detected and Masson staining was performed;toluidine blue staining of the distal nerve injury and immunofluorescence staining of the damaged nerve were performed.
RESULTS AND CONCLUSION:At 4, 8, 12 weeks after cel implantation, the sciatic nerve function index was ranked as fol ows:frozen nerve injury group

Objective: To investigate the clinical effect of free medical sural artery perforator myocutaneous flap for repairingof the woundwith osteomyelitis. Methods: 17 patients suffered from the wound with osteomyelitis were treated in the Ningbo First People′s Hospital, There were 11 males and 6 females with an mean age of 53.2 years (range, 21-76 years). The sizes of the defect ranged 5 cm×4 cm to 13 cm×6 cm. All patients underwent debridement and used antibiotic-loaded bonecement to cover the wound. Meanwhile, patients were treated with sensitive antibiotics, operation and free medical sural artery perforator myocutaneous flap were used to treat the wound. Preoperative use ultrasound and CT angiography to positioning perforator, The flap area ranged from 6 cm×4 cm to 13 cm×7 cm and the donor sites were closed directly. The author provided the patients with the treatment of anti-inflammatory, anti-spasmodic and anti-coagulantin the postoperative. Used infrared thermograms to assess the flap blood supply. Results: One flap skin margin was non union due to poor blood supply.All of the other 16 flaps success survived and the donor sites were closed directly. Postoperative follow-up period was 4 to 23 months and the flaps had satisfied texture and appearance.All the donor sites had a good healing with no pain and complications, also the osteomyelitis was controlled. Conclusions The free medical sural artery perforator myocutaneous flap is reliable for reconstruction of the wound with osteomyelitis.

Objective:To explore the feasibility and clinical effectiveness of preoperative positioning grid combined with CTA assisted design of anterolateral thigh perforator flap (ALTPF) in reconstruction of soft tissue defects around foot and ankle.Methods:From May 2018 to December 2021, a total of 18 cases with soft tissue defects around foot and ankle were admitted to the Department of Plastic and Reconstructive Surgery, First Affiliated Hospital of Ningbo University. The patients were 11 males and 7 females, aged from 32 to 78 years old, with an average age of 57.5 years old. Among them, 10 had chronic ulcer wounds, 6 had traumatic wounds, and 2 had postoperative wounds after malignant tumor resection. The sizes of soft tissue defect ranged from 6.0 cm×8.0 cm to 9.0 cm×13.0 cm, and the flap sizes ranged from 8.0 cm×10.0 cm to 11.0 cm×15.0 cm. Preoperative positioning grid combined with CTA three-dimensional reconstruction were used to mark the exit point of the perforator vessels in digital format, in order to restore the course of vessels and calculate the length of the vascular pedicles. ALTPFs were accurately designed based on the digitally reconstructed images, and then the ALTPFs were used to repair the soft tissue defects around the ankle. The flap donor sites were directly closed in stage Ⅰ. After the operation, all the patients were included in scheduled follow-ups at the outpatient department to observe the appearance of the recipient flaps and donor sites. The functional evaluation of the affected feet were assessed according to the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score.Results:All 18 flaps survived, and the wounds in both the recipient and donor sites healed in stageⅠ. Postoperative follow-up for the 18 patients were 5 to 36 months, with an average of 13 months. At the last follow-up, the appearances of the flaps were good without swelling, the donor sites had good recovery, and there was no obvious scar hypertrophy. The function evaluation of the affected feet were found at excellent in 10 cases, good in 6 cases, and fair in 2 cases.Conclusion:Preoperative position grid combined with CTA three-dimensional reconstruction can digitally mark the exit points and running courses of the perforator vessels. It is an effective method for accurate position of perforator vessels before surgery. It can effectively reduce the operation time, lower the surgical risks, and achieve a high survival rate of the flap, thus holding considerable clinical value.