Potency is one of the most important indexes of inactivated vaccines.A number of methods have been established to assay the potency,of which the NIH test and single-dose mouse protection test are the prescribed methods.Here,we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine,which uses fewer animals and takes less time to complete.Depending on the quality requirements of a vaccine(e.g.minimum potency),a rabies reference vaccine is,for example,diluted to the minimum potency,and 50 μL of the dilution is taken to inoculate 10 mice.The same amount of the test rabies vaccine is inoculated into another 10 mice.After two weeks,all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization(FAVN) test.By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine,the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality.The reliability of this method was also confirmed in dogs.The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.

Objective To analyze the treatment principles of burn patients with fractures and the clinical application of external fixation.Methods The treatment methods and efficacy in 48 patients with bum complicated fracture were summarized.Limb fractures were 53,femoral shaft 21,tibia and fibula 28,humerus 13,ulnar bone torsion 5.Closed 44,open 23.All limbs received external fixation,with single-arm multifunctional external fixator in 47,bilateral 17,Monteggia 3.Burn wound dressing was dealed with the situation in accordance with surgical debfidement or skin flap.Results The 48 cases were found no deaths,and 67 fractures healed and discharged healing time 2.5～5 months,3.1 months on average.Limb and joint function showed a good recovery.Conclusions External fixation showed fixation stability,simple operation,mild tissue injury,easy handling and anti-burn scar contracture and other advantages.Clinical results were satisfactory.

OBJECTIVE:To evaluate the clinical efficacy and safety of letrozole combined with traditional Chinese medicine for ovulation dysfunction induced by polycystic ovarian syndrome(PCOS). METHODS: 300 PCOS outpatients were randomized to 3 groups of 100 each: clomiphene citrate (CC) group (188 cycles);urinary gonadotropin (HMG) group (104 cycles) and letrozole (LE) group (155 cycles). The occurrence rate of mature follicle,ovulation rate,pregnancy rate,abortion rate,superfoetation-terata rate,cancellation rate,incidence of no dominant follicle forming and OHSS etc were compared between the two groups after treatment. RESULTS: 30 babies were born in CC group,34 in HMG group and 43 in LE group were born. The occurrence rate of mature follicles,ovulation rate and pregnancy rate in the LE group were higher than in either CC group or HMG group(P0.05);the superfoetation rate in HMG group was higher than in CC group or LE group(P

Objective Clinical effects of dot from body skin irradiated xenogeneic skin improved composite graft.Methods Selected 80 patients with severe burn patients randomized after admission on the basis of conventional treatment,3-5 days line Crust treatment group transplanted autologous point-like skin irradiation pigskin coverage.Results Treatment of patients in group one week dressing see irradiated pigskin Tiefu full two weeks the pigskin was dry scab-like.Wound healing treatment group (29 ± 5) days (P ＜ 0.01) was significantly shorter than the control group(39 ±4) days.Wound healing rate of treatment is significantly higher than the control group (P ＜0.01).Wound infection rate reduced greatly reduce labor intensity and dressing,and reduce pain.1 year after scar formation in patients with good flexibility and functionality.Conclusions Punctate since improved composite body skin irradiated xenogeneic skin transplantation can improve skin graft survival rate,promote wound healing,treatment of a large area of the burn wound repair is feasible and effective.

BACKGROUND:Previously deep burn wound or skin defects are generaly repaired with skin grafting or flap of skin grafting. Obvious scar hyperplasia usualy appears after operation, which requires multiple surgeries. Meanwhile, patients have to suffer from great pain and bear high cost. OBJECTIVE: To observe the clinical effects on deep wounds by continuous traction of self-designed skin-stretching device (patent No. ZL 2012 2 0022443.7). METHODS: Thirty patients with deep burn wound, skin defect or funicular scar were enroled, including 22 males and 8 females, aged 18-49 years, and randomly divided into two groups. Skin-stretching device was adopted for skin traction treatment. Twenty cases underwent skin traction from 1 kg puling force to 5 kg, with an increase of 1 kg per 2 days, 6 hours a day for 10 days. Blood flow at the beginning, 1, 5, 10, 15, 20, 30, 60 minutes of the skin traction, and the changes of wound edge skin as wel as histological changes of the skin were observed. Of the remaining 10 cases, 2, 6, and 2 cases underwent skin traction of 2, 4, 7 kg, respectively. Blood flow and skin changes were also observed to find out the most suitable and safe force. RESULTS AND CONLUSION:Al the 30 cases achieved primary healing without necrosis of skin, infection or peripheral circulatory disorders, and the appearance and function recovered wel. The healing time was 8-24 days. The skin-stretching device was most safe under 4 kg puling force, by which, there was neither blood circulation obstacle nor tear of skin. After traction, the skin blood flow and the number of cels increased, especialy the epithelial basal cels. The colagen fibers became thicker and denser, and the elastic fibers regenerated significantly; the fibroblasts and capillary density increased. It has been proved that we can better close the wound and reduce scar formation effectively with the self-designed skin-stretching device for skin traction.

Objective To investigate the efficacy and safety of compound lysostaphin disinfectant solution in the latter stage of the treatment of bum wounds infection,and to evaluate its clinical value.Methods The selected 60 cases of bum wound infection were grouped into treatment and control group at random.The treatment group recieved compound lysostaphin disinfectant solution(Shanghai Hi-tech Co.bio-engineering,Ltd.)with debridement and wet,once a day;the control group with normal saline+gentamicin wet gauze,once a day.After cleaning the wound two groups 40 cases of underwent regular skin debridement,20 cases in each group.The bacteria changes in the wound were detected,of bacteria removal the accumulated rate of wound healing time and healing rate were compared.Results 3,6,9 days after treatment the accumulated rate of bacteria removal was 45.8%,73.9%, 89.5%higherthan the control group(13.8%,27.6%,57.4%)respectively(P＜0.01).Wound healing time in the treatment group was(12±5)d,shorter than the control group(16±4)d Was significantly(P＜0.01). The rate of wound healing in the treatment group was significantly higher(P＜0.01).No adverse reactions were found.Conclusion The treatment application of compound iysostaphin disinfectant in the later stage of bum wound is effective in control of infection,increases the survival rate of skin and shortens wound healing time,so it is safe and effective.

Objective To assess the value of four different techniques of detecting the Mycobacterium tuberculosis (MTB) in bronchoalveolar lavage fluid (BALF) in the diagnosis of tracheobronchial tuberculosis. Methods A total of 98 patients diagnosed as tracheobronchial tuberculosis were selected from May 1,2013 to June 30,2016. The clinical data was analyzed retrospectively,and the positive rates of MTB of the 960 cultrue, the direct smears , the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were compared. Results The positive rates of the 960 cultrue,the direct smears,the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were 20.4%(20/98),15.3%(15/98),70.4%(69/98) and 74.5%(73/98),respectively. Among the four techniques ,the positive rates of the modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay were significantly higher than those of the 960 cultrue and the direct smears(P<0.05,respectively). However,no significant difference was found between the modified Ziehl?Neelsen stain method and the Xpert MTB/RIF assay (P > 0.05). Conclusions The modified Ziehl?Neelsen stain method and Xpert MTB/RIF assay for detecting the MTB in BALF have high clinical value in the diagnosis of tracheobronchial tuberculosis.

Objective To test the immune therapeutic effects of PICKCa adjuvant rabies vaccine for the post-exposure mice and beagle dogs and immune memory effect for human.Methods For the mice and beagle dogs first to infect with rabies wild virus and then immunize with PICKCa rabies vaccine.For volunteers, first to immunize with PICKCa rabies vaccine after 2.5 years to take peripheral blood mononuclear cell and then to check cells secreted interferon-gwith flow cytometry. Results In 3 independent assays of post-explosure immunization of mice and beagle dogs PICKCa rabies vaccine were much better then commercial adjuvant-free rabies vaccines, the protective rate from 20%-30%to 70%-100%Also, in the assay of cells secreted IFN-γby peripheral blood mononuclear cells from volunteers immunized by PICKCa rabies vaccine the IFN-γcell levels were much higher than control.Conclusions Compared with commercial adjuvant-free rabies vaccines the PICKCa rabies vaccine had significant immune therapeutic effects in the mice and dogs.Also PICKCa rabies vaccine had good immune memory effect in human.

African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.

Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation.Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group,PD-L1 inhibitor group,pirfenidone group and combined treatment group(n=10).After successful modeling,PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days,and oral pirfenidone(500 mg/kg)was given on a daily basis.The survival rate of the mice and tumor growth rate were compared among the 4 groups.The expressions of CD3,CD8,CD45,E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment,and bone marrow-derived suppressor cells(MDSCs)were observed with immunofluorescence staining;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN),creatinine(CRE)and lactate dehydrogenase(LDH-L)were analyzed using an automated biochemical analyzer.Results Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days(P<0.05),but the combined treatment produced an obviously stronger inhibitory effect(P<0.05).PD-L1 inhibitor and pirfenidone alone significantly increased E-cadherin expression and decreased N-cadherin expression in the tumor tissue(P<0.05).The two treatments both significantly increased the percentage of CD3+,CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue,and these changes were more obvious in the combined treatment group(P<0.05).No significant differences were found in serum ALT,AST,BUN,CRE or LDH-L levels among the 4 groups(P>0.05).Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.