OBJECTIVE:To establish a method for determination of the principle components in streptomycin sulfate in Streptomycin sulfate/cod liver oil nasal drop.METHODS:UV spectrophotometry was use to determine the absorption of streptomycin sulfate at detection wavelength of 521 nm.The content of streptomycin sulfate was also calculated.RESULTS:The detection concentration of streptomycin sulfate has favorable linear correlation within 250～700 ?g?mL-1 (r=0.999 8).The average recovery was 99.75% (RSD=0.98%).CONCLUSION:The method is convenient,accurate and suitable for quality control of the preparation.

ObjectiveTo explore the tumor igenic property of side population cells (SP) from human gallbladder carcinoma cell line GBC-SD. Methods SP and non-SP cells were isolated from GBC-SD staining with Hoechst33342 dye by fluorescence-activated cell sorting (FACS). The soft agar clonal assay and xenograft assay were performed to characterize tumorigenic property of side population cells in vitro and in vivo, respectively. The percentage of SP cells was analyzed by FACS in 5 hu man gallbladder carcinoma specimens. ResultsThe percentage of SP cells accounted for approximately 0.87 ％ of GBC-SD cells. The clone-formed rates of SP was more frequent than that of non-SP cells (14.74％ ± 3.53％ vs 5.17％ ± 1.05％), there was statistically significant difference (t =2.75,P＜0. 05). SP cells could generate tumors with as few as 5 × 103 cells (four of seven animals), whereas at least 1 × 105 non-SP cells were needed to form a tumor (one of seven animals). Re-analysis of SPderived tumors by FACS showed that SP cells under in vivo conditions also have the capacity to regenerate the SP and non-SP fractions. Besides, analysis of Hoechst33342 revealed s small fraction of SP cells, ranging from 0. 27％ to 2.3％ in gallbladder carcinoma specimens. ConclusionSP cells from GBC-SD are highly tumorigenic similar as the cancer stem cells.

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.

Objective To discuss the feasibility and clinical application of percutaneous internal fixation (PIF) combined with pecutaneous osteoplasty (POP) for the treatment of metastasis of proximal femur with impending pathological fracture. Methods Six consecutive patients with metastases of the proximal femur, who could not be able to tolerate conventional surgery, underwent PIF together with POP. The results were analyzed. Results The procedure was successfully accomplished in all six patients. Neither pulmonary embolism nor death occurred in all patients during and after the operation. No fracture at operated area was observed during follow-up period. Conclusion For stabilization of proximal femoral metastatic lesion with impending pathological fracture, percutaneous internal fixation combined with pecutaneous osteoplasty is a safe and effective technique.

Objective To discuss the feasibility of biodegradable covered magnesium alloys stent for a rabbit model of lateral aneurysm in the common carotid artery (CCA). Methods The left CCA was ligated in 20 conventional New Zealand rabbits. MRA of the neck was performed 1 month after ligation. The rabbits with thickening of the right CCA and non-or slight thickening of the bilateral vertebral arteries were selected for lateral aneurysm model making. The venous pouch and the right CCA wall by discontinuous extroversion sutures, to form a lateral aneurysm model. The biodegradable covered magnesium alloys stents or Willis covered stents were inserted two weeks after model making. Angiography was performed prior to the procedure, 3, 6 and 12 months after stent implantation to evaluate the disappearance of the aneurysms and patency of the right CCA. The degradation behaviour is invastagated with molybdenum target 2 weeks, 1, 2, 4, 6,9 and 12 months after stent placement. Results The left CCAs were successful ligated in all rabbits. MRA 1 month after ligation showed thickening of the right CCA and non-thickening of the bilateral vertebral arteries in 17 of 20 rabbits. In these animals, the CCA lateral aneurysm model was deemed successful, and biodegradable covered magnesium alloys stents and Willis covered stents were implanted in 9 and 8 aneurysms, respectively. DSA after biodegradable covered magnesium alloys stent placement displayed disappearance of the aneurysms and patency of the CCA in all 9 rabbits during follow-ups. DSA 3 months after Willis stent placement displayed patency of the CCA in 7 rabbits and occlusion of the artery in one. No occlusion of the right CCA was observed on angiography at 6 and 12 months. The degradation of the biodegradable covered magnesium alloys stent was investigated with molybdenum target during follow-ups, and no changes was observed in Willis covered stent. Conclusion Biodegradable covered magnesium alloys stent is a feasible approach for the treatment of a lateral aneurysm in the right CCA.

Objective To investigate the clinical efficacy of open reduction and internal fixation with dynamic anterior plate-screw system for quadrilateral area ( DAPSQ ) in the treatment of elderly patients with acetabular fracture involving medial displacement of the quadrilateral surface. Methods Between January 2007 and December 2014, a series of 18 senile patients with acetabular fractures involving medial displace-ment of the quadrilateral surface were treated at our department. They were 13 men 5 women, with a mean age 67. 8 years ( range, from 61 to 78 years ) . By the Judet-Letournel classification, there were 4 anterior column fractures, 5 anterior column plus posterior hemitransverse fractures, 8 double column fractures and one T-shaped fracture. The delay from injury to surgery averaged 6. 5 days. All were treated with the technique of DAPSQ. Screw internal fixation of the quadrilateral area followed plate moulding via a single ilioinguinal ap-proach. Fixation with lag screws for the posterior column was added if necessary. Results The 18 pa-tients were followed up for 12 to 35 months ( mean, 26 months ) . By the Matta criteria for fracture reduction, 9 cases were rated as excellent, 6 as good and 3 as poor. Bone healing was achieved within 2 to 4 months( mean, 3. 5 months ) . By the modified Merle d'Aubigné & Postel criteria, the affected hips scored 9 to 18 points ( mean, 16. 3 points ) at the final follow-ups, giving 7 excellent, 7 good, 2 fair and 2 poor cases. By the Harris evaluation, the affected hips scored 58 to 98 points ( mean, 87. 6 points ) , giving 7 excellent, 8 good, one fair and 2 poor cases. The postoperative complications included urinary tract infection in one, lesion of lateral femoral cutaneous nerve in one, heterotopic ossification ( Brooker Grade Ⅰ) in one, and traumatic osteoarthritis in 5 cases 2 of whom had to receive total hip arthroplasty. Conclusion The technique of DAPSQ is suitable for the treatment of acetabular fractures involving medial displacement of the quadrilateral surface in elderly patients, because it can provide and maintain stable fixation of the fracture in the quadri-lateral area and preserve hip joint function, leading to fine outcomes.

Objective To discuss the feasibility and short-term clinical effectiveness of DSA-guided percutaneous vertebroplasty (PVP) for the treatment of painful osteoblastic metastatic spinal lesions. Methods During the period from Jan. 2010 to Dec. 2011 at authors’ hospital PVP was carried out in 23 patients with osteoblastic spinal metastases (34 lesions in total). Coexisting osteoblastic pathological fracture was found in twelve patients. The WHO standards, visual analogue scale (VAS) and karnofsky-KPS score were used to evaluate the therapeutic results. Results Technical success was achieved in all patients. All patients were followed up for at least 3 months. Of 20 patients who had complete clinical data, complete remission (CR) was obtained in 6, partial remission (PR) in 10, mild remission (MR) in 3 and no remission (NR) in one. The clinical effectiveness (CR+PR) was 80%. The mean VAS scores dropped from preoperative (7.0 ± 1.6) to (2.2 ± 1.9) at 24 hours after the treatment, and to (2.4 ± 2.1) and (2.5 ± 2.1) at one and three months after the treatment respectively. The mean KPS scores rose from preoperative (76.5 ± 10.4) to (86.5 ± 11.8), (88.0 ± 12.0) and (89.0 ± 10.8) at 24 hours and one, three months after the treatment respectively. Small amount leakage of PMMA was observed in 4 cases (17.4%) with no obvious clinical symptoms. Conclusion DSA-guided PVP is a feasible and effective treatment for painful osteoblastic spinal metastases. This therapy can immediately relieve pain and reinforce spine, besides, it can remarkably improve the living quality and
decrease the incidence of paraplegia.

Objective:To investigate the effect of percutaneous kyphoplasty on the vertebral height and Cobb angle in elderly patients with osteoporotic thoracolumbar compression fractures.Methods:Seventy elderly patients with osteoporotic thoracolumbar compression fractures who were admitted to the Affiliated Hospital of Hubei University of Traditional Chinese Medicine from March 2019 to March 2020 were selected as the study objects. They were grouped according to the random number table method, with 35 patients in each group. The patients in the observation group were treated with multi-point balloon expansion percutaneous kyphoplasty, and the patients in the control group were treated with single balloon expansion percutaneous kyphoplasty. The Visual Analogue Scale (VAS) and Oswestry Dysfunction Index (ODI) score were compared between the two groups before and after treatment. The relative height of injured vertebrae, Cobb angle, bone cement diffusion volume ratio, operation time, radiation exposure time and bone cement injection volume were recorded.Results:There was no significant difference in VAS and ODI score between the two groups before operation (all P>0.05). At 12 months after operation, the VAS and ODI score of the two groups were lower than those before operation (all P<0.05), and the ODI score of the observation group was significantly lower than that of the control group ( P<0.05). At 12 months after operation, the relative height of injured vertebrae in the observation group was higher than that in the control group, and the local Cobb angle was significantly lower than that in the control group (all P<0.05). The total effective rate of the observation group was significantly higher than that of the control group (94.28% vs 82.86%, P<0.05). Compared with the control group, the observation group had higher proportion of grade Ⅱ in the diffusion volume ratio of bone cement and more bone cement injection, and longer operation time (all P<0.05), while there was no significant difference in the radiation exposure time between the two groups ( P>0.05). Conclusions:The treatment of percutaneous kyphoplasty with multi-point expansion of balloon under overextended posture can more effectively improve the relative height of injured vertebrae and improve local Cobb angle of elderly patients with osteoporotic thoracolumbar compression fractures, and does not increase the risk of bone cement leakage. It is worthy of clinical application and promotion.

Objective To investigate the effect and mechanism of Qianggu Tongbi capsule on osteoarthritis factors in rats.Methods Osteoarthritis(OA)rat model was established by anterior cruciate ligament transection method.Rats were ran-domly divided into model group,Qianggu Tongbi capsule group and glucosamine group.HE staining was used to identify the model.Toluidine blue staining was used to observe the changes of cartilage.Real-time quantitative polymerase chain reaction(qRT-PCR)was applied to detect the expression of Toll-like receptors(TLR4)and nuclear factor kappa-B(NF-κB)in each group.The levels of IL-1β,TNF-α,and IL-6 were measured by the enzyme linked immunosorbent assay(ELISA)kit.The levels of TLR4 and p-NF-κB were detected by immunohistochemistry.Results HE staining showed that,compared with the control group,a large number of inflammatory cell infiltration and synovial tissue thickening were observed in the model group,while the inflammatory cell infiltration and synovial tissue thickening were not obvious in the Qianggu Tongbi capsule and glu-cosamine groups.Toluidine blue staining results showed that chondroitin sulfate in the cartilage tissue of the model group was significantly reduced compared with the control group,while levels of chondroitin sulfate in the Qianggu Tongbi capsule group and glucosamine group were significantly increased.Compared with control group,the expression and immunohistochemical re-sults of TLR4 and NF-κB,and the levels of IL-1β,TNF-α and IL-6 in model group were significantly increased(all P＜0.05).Compared with model group,the expression and immunohistochemical results of TLR4 and NF-KB,and the levels of IL-1β,TNF-α and IL-6 in Qianggu Tongbi capsule group and glucosamine group were significantly decreased(all P＜0.05).Conclusion Qianggu Tongbi capsule can inhibit expression of TLR4 and NF-κB by reducing the production of inflammatory factors to a-chieve the effect of treatmenting osteoarthritis.

Objective:To investigate the immune effects of Clostridium difficile toxoid B (CdtB) vaccine formulated with different mucosal adjuvants through microneedle immunization, and to provide ideas for the prevention and treatment of Clostridium difficile infection. Methods:CdtB vaccine was prepared with purified Clostridium difficile toxin B(TcdB) after formaldehyde detoxification. Female BALB/c mice were immunized with different doses of vaccine alone or in combination with mucosal adjuvants. The titers of specific serum IgG and fecal IgA were detected at 0 d, 7 d, 14 d, 28 d and 42 d after immunization. The protective effects of CdtB vaccine were evaluated by cell neutralization assay and Clostridium difficile challenge infection. Results:(1) With the increase of immune dose, the mice immunized with CdtB vaccine alone by microneedle not only produced better serum specific IgG, but also had higher level of IgA in feces. (2) When the mice were immunized with CdtB vaccine containing LT or CTB adjuvant by microneedle, the trend of serum specific IgG titer in each group increased with the increase of immune dose, especially in the group containing LT adjuvant. There were significant differences in the trend of specific IgA titer in feces between the adjuvant groups and the group without adjuvant, but the adjuvant effect was not obvious. (3) No significant difference in serum IgG titer was observed between the mice immunized with 10 μg CdtB by microneedle or intraperitoneal injection, but microneedle immunization significantly increased fecal IgA level. (4) The neutralization titers of specific antibodies in mouse serum after immunization and the test results of challenge protection in mice confirmed that the use of CdtB vaccine had certain protective effects.Conclusions:CdtB vaccine had better immune effects in mice through microneedle immunization, but the adjuvant effects of LT and CTB were not significant.