Effect evaluation of denosumab combined with curettage and bone cement reconstruction in the treatment of recurrent giant cell tumor of bone around the knee joint.

OBJECTIVE: Giant cell tumor of bone (GCTB) is a common primary bone tumor with latent malignant tendency. GCTB is prone to occur around the knee joint, and surgery is the major treatment method. There are relatively few reports on denosumab in the treatment of recurrent GCTB around the knee joint and postoperative function evaluation of patients. This research aimed to explore the appropriate surgical options for the treatment of recurrent GCTB around the knee joint. PATIENTS AND METHODS: 19 patients with recurrent GCTB around the knee joint, who were admitted to Hospital for 3 months following denosumab treatment from January 2016 to December 2019, were included as the research subjects. The prognosis was compared between patients treated with curettage combined with polymethylmethacrylate (PMMA) and those with extensive-resection replacement of tumor prosthesis (RTP). A deep learning model of Inception-v3 combined with a Faster region-based convolutional neural network (Faster-RCNN) was constructed to classify and identify X-ray images of patients. The Musculoskeletal Tumor Society (MSTS) score, short form-36 (SF-36) score, recurrence, and the rate of complications were also analyzed during the follow-up period. RESULTS: The results showed that the Inception-v3 model trained on the low-rank sparse loss function was obviously the best for X-ray image classification, and the classification and identification effect of the Faster-RCNN model was significantly better than that of the convolutional neural network (CNN), U-Net, and Fast region-based convolutional neural network (Fast-RCNN) models. During the follow-up period, the MSTS score in the PMMA group was significantly higher than that in the RTP group (p<0.05), while there was no significant difference in the SF-36 score, recurrence, and the rate of complications (p>0.05). CONCLUSIONS: The deep learning model could improve the classification and identification of the lesion location in the X-ray images of GCTB patients. Denosumab was an effective adjuvant for recurrent GCTB, and widely extensive-resection RTP could reduce the risk of local recurrence after denosumab treatment for recurrent GCTB.

[Summary and analysis of total auricle reconstruction in adult microtia patients].

Objective: To observe the clinical effect of auricle reconstruction in adult patients with microtia and summarize the experience. Methods: Clinical data of adult patients with microtia who underwent total auricle reconstruction using the modified Nagata's two stage for microtia reconstruction from June 2016 to June 2021 were analyzed. A total of 41 adult patients (42 ears) with microtia were enrolled, including 30 males and 11 females, with the median age at the time of surgery of 37 years. Autogenous costal cartilage was used as the auricular framework for all patients in this group. The first stage surgery was performed according to the modified Nagata's two stage for microtia reconstruction procedure,cartilage auricular framework carving was performed by different methods according to the ossification state of adult costal cartilage. Six months following the primary operation, ear elevation and cranioauricular angle formation, retroauricular facial flap transfer and medium-thick skin grafting were performed in the second stage. Results: All patients successfully completed two stage operation. During the follow-up of 3 months and 24 months, all the 41 patients were satisfied with the morphology of reconstructed auricle. Conclusion: According to the costal cartilage status of adult patients, different costal cartilage carving techniques can be used for total auricle reconstruction to obtain ideal surgical results.

[Effects and mechanism of interleukin-17-modified mouse bone marrow mesenchymal stem cells on rejection reaction of allogeneic skin transplantation in mice].

Objective: To explore the effects and mechanism of interleukin-17 (IL-17)-modified mouse bone marrow mesenchymal stem cells (BMSCs) on the allogeneic skin transplantation in mice. Methods: (1) The femur, tibia, and humerus were isolated from five BALB/c mice (all female, aged 4 to 8 weeks, the same gender and age below) after sacrifice. BMSCs were isolated, purified, and cultured by whole bone marrow density gradient centrifugation combined with adherent separation method. The third passage of cells was used for morphological observation and identification of adipogenic and osteogenic differentiation. The fourth passage of cells was used for identification of the expression of stem cell surface markers. The third to sixth passages of BMSCs were pretreated with mouse recombinant IL-17 at a final mass concentration of 50 ng/mL for 5 days, and then were harvested for morphological observation. After being labeled with carbocyanine fluorescent dye (CM-Dil), IL-17-pretreated BMSCs and IL-17-unpretreated BMSCs were obtained for morphological observation and the labeling rates were calculated. (2) Forty-five C57BL/6J mice were divided into phosphate buffer solution (PBS) control group (n=13), BMSCs alone group (n=16), and BMSCs+ IL-17 group (n=16) according to the random number table. One day before the skin transplantation of mice, 0.1 mL BMSCs (5×10(6) cells/mL) without CM-Dil labeling were injected to the 13 mice in BMSCs alone group through the tail vein, and 0.1 mL BMSCs (5×10(6) cells/mL) labeled with CM-Dil were injected to the other 3 mice in BMSCs alone group through the tail vein. IL-17-pretreated BMSCs (5×10(6) cells/mL) without CM-Dil labeling in the volume of 0.1 mL were injected to the 13 mice in BMSCs+ IL-17 group through the tail vein, and 0.1 mL IL-17-pretreated BMSCs (5×10(6) cells/mL) labeled with CM-Dil were injected to the other 3 mice in BMSCs+ IL-17 group through the tail vein. PBS in the volume of 0.1 mL was injected to the 13 mice in PBS control group through the tail vein. Forty-five BALB/c mice were used as donors, and forty-five treated C57BL/6J mice in the 3 groups were used as recipients to establish a back-to-back full-thickness skin transplantation model. On the 2nd day after transplantation, the same number of corresponding cells and the equal amount of PBS were injected to the recipient mice of each group again. On the 7th day after transplantation, three mice injected with CM-Dil-labeled BMSCs in BMSCs alone group and three mice injected with CM-Dil-labeled IL-17-pretreated BMSCs in BMSCs+ IL-17 group were sacrificed by cervical dislocation to track the CM-Dil-labeled BMSCs by fluorescence microscope, which was counted. After the dressing removal on the 6th day post transplantation, 7 mice were selected respectively from 13 mice in BMSCs alone group injected with BMSCs without CM-Dil-labeling, 13 mice in BMSCs+ IL-17 group injected with IL-17-pretreated BMSCs without CM-Dil-labeling, and 13 mice in PBS control group, respectively, to record the skin graft survival time. On the 8th day post transplantation, three of the remaining six mice in the three groups were taken for general observation of the grafted skin, serum levels of interferon-γ, IL-10, and transforming growth factor ß (TGF-ß) by enzyme-linked immunosorbent assay method, the percentage of CD4(+) CD25(+) forkhead/winged helix transcription factor p3 (Foxp3)(+) regulatory T cells (Tregs) in spleen by flow cytometer, and the histopathological observation of the grafted skin by hematoxylin eosin staining. The rest three mice in each group were also taken for histopathological observation as above on the 14th day post transplantation. Data were statistically analysed with independent sample t test, one-way analysis of variance, and least significant difference test. Results: (1) There were no significant differences in the morphology and size between IL-17-pretreated BMSCs and IL-17-unpretreated BMSCs on culture day 5. (2) After CM-Dil labeling, BMSCs and IL-17-pretreated BMSCs grew well, and the labeling rate was almost 100%. (3) On the 7th day post transplantation, there were 6.2±2.6 CM-Dil-labeled BMSCs per 100 fold visual field in the skin and adjacent subcutaneous tissue of mice in BMSCs alone group, which were significantly fewer than the 15.0±5.3 CM-Dil-labeled IL-17-pretreated BMSCs per 100 fold visual field in BMSCs+ IL-17 group (t=-2.962, P<0.05). (4) The skin graft survival time of mice in BMSCs alone group and BMSCs+ IL-17 group was (13.3±1.2) and (17.0±1.5) days respectively, significantly longer than (8.7±0.8) days in PBS control group (P<0.01), and the skin graft survival time of mice in BMSCs+ IL-17 group was significantly longer than that in BMSCs alone group (P<0.01). (5) On the 8th day post transplantation, most of the skin grafts of mice in PBS control group was black, scabby, and necrotic. Most of the skin grafts of mice in BMSCs alone group survived well, while all the skin grafts of mice in BMSCs+ IL-17 group survived well. (6) On the 8th day post transplantation, compared with those of PBS control group, the serum levels of IL-10 and TGF-ß of mice in BMSCs alone group and BMSCs+ IL-17 group were significantly higher (P<0.01), and the serum level of interferon-γ was significantly lower (P<0.01). Compared with those of BMSCs alone group, the serum levels of IL-10 and TGF-ß of mice in BMSCs+ IL-17 group were significantly higher (P<0.01), and the serum level of interferon-γ was significantly lower (P<0.01). (7) On the 8th day post transplantation, the percentages of CD4(+) CD25(+) Foxp3(+) Treg in spleen of mice in BMSCs alone group and BMSCs+ IL-17 group were significantly higher than the percentage of PBS control group (P<0.01), and the percentage of CD4(+) CD25(+) Foxp3(+) Treg in spleen of mice in BMSCs+ IL-17 group was significantly higher than that of BMSCs alone group (P<0.01). (8) On the 8th day post transplantation, infiltration of a large number of inflammatory cells and necrosis of epidermis and dermis were found in the skin grafts of mice in PBS control group; focal infiltration of inflammatory cells and slight epidermal degeneration were found in the skin grafts of mice in BMSCs alone group; the skin appendages of the skin grafts of mice in BMSCs+ IL-17 group survived well with angiogenesis. On the 14th day post transplantation, the skin grafts of mice in BMSCs alone group showed extensive infiltration of inflammatory cells, severe epidermal degeneration and focal necrosis; the skin grafts of mice in BMSCs+ IL-17 group showed focal infiltration of inflammatory cells and slight epidermal degeneration; the skin grafts of mice in PBS control group were completely necrotic. Conclusions: IL-17 can reduce the immune rejection in allogeneic skin grafting and prolong the survival time of mouse skin grafts by improving mice BMSCs' capabilities to induce immune tolerance and enhancing the homing ability of BMSCs.

[Antagonistic effect of oleanolic acid on anaphylactic shock].

AIM: To study the antagonistic effect of oleanolic acid (OA) on anaphylactic shock in guinea-pigs sensitized with 6-aminopenicillanic acid (APA)-protein. METHODS: The anaphylactic shock was achieved by i.p. of APA-protein in guinea pigs. The serum antibody titers of guinea pigs were measured with the passive cutaneous anaphylaxis test, and the pulmonary histamine by the method of fluorometric determination. The degranulation of mast cells was examined by rat calvarian periosteum method. RESULTS: In guinea pigs sensitized with APA-protein, OA 25, 50, 100 mg . kg-1 reduced anaphylactic shock rates from 100% (sensitized control group) to 70%, 18%, and 15% (OA pretreated group), and death rates from 100% to 70%, 18%, and 0%, respectively. OA had an inhibitory effect on the antibody formation and reduced the histamine contents in the lung tissues in the sensitized guinea pigs. In addition, the homologous passive cutaneous anaphylaxis in guinea pigs and the degranulation of mast cells of calvarial periosteum in rats were prevented by OA 25, 50, and 100 mg . kg-1. CONCLUSION: The antagonistic effect of OA on anaphylactic shock was related to the inhibition of antibody formation and the reduction of histamine contents.

[Influence of carbenicillin on pharmacokinetics of ribostamycin in 7 volunteers when they were used combinedly].

The experiment consists of two groups, namely the ribostamycin (Rib) group and the Rib and carbenicillin (Car) combination group. There were 7 healthy volunteers in each group. The Rib concentration in blood and urine was detected with the improved microbiology method. The organism tested was Bacillus pumilus. The results demonstrated that the drug serum level was in line with the one-compartment open model. The parameters such as K, Ka, T1/2, T1/2a, Tp, Cmax, and AUC in the Rib group and the combination group were 0.43 +/- 0.03 and 0.41 +/- 0.04 h-1, 3.09 +/- 1.37 and 2.82 +/- 1.48 h-1, 1.61 +/- 0.12 and 1.72 +/- 0.17 h, 0.26 +/- 0.12 and 0.31 +/- 0.15 h, 0.91 +/- 0.20 and 1.01 +/- 0.31 h, 37 +/- 4 and 34 +/- 6 micrograms.ml-1, 123 +/- 17 and 120 +/- 16 micrograms.h.ml-1, respectively. The 24-h urinary recovery rates of the 2 groups were 84 +/- 3% and 84 +/- 5%, respectively. There was no significant difference (P greater than 0.05) between the parameters of the 2 groups. The results indicate that Car has no apparent effect on the pharmacokinetics of Rib in the healthy volunteers.

Distribution of juxtaglomerular cells and synthesis and release of renin granules in patients with renal artery stenosis.

100 pieces of renal tissue from 4 patients with renal artery stenosis were studied histologically. Juxtaglomerular cells were found not only in the region close to the glomerulus but also in the proximal 2/3 of the afferent arteriole of the glomerulus, but not in the interlobular arterioles. In the cytoplasm of juxtaglomerular cells, three kinds of granules were found: crystalline granules (immature), the round or ovoid granules (mature), and the lobulated granules (fusion). Renin was synthesized in the rough endoplasmic reticulum and packaged in the Golgi complex. Electron microscopic findings suggested that the secretory microvesicles were produced on the surface of Golgi complex. They fused to form primary crystalline granules, which were enlarged by renin supplied by transportation vesicles and finally formed typical crystalline granules. The mature renin granules (round or ovoid) moved toward the periphery of the cytoplasma. Their limiting membranes on the side toward the basement membrane disintegrated and formed microvesicular bodies passing through the periplasm, and the basement membrane and were discharged into the interstitial matrix.

[Changes in degranulation and histamine release from mast cells in rats sensitized by penicilloyl-protein].

In this paper, the mechanism of the penicillin G anaphylactic reaction was studied. Rats were sensitized to penicilloyl (BPO)-protein. The incidences of anaphylactic reaction and death were 100 and 40%, respectively. After being challenged, the degranulation rates of rat cranial periosteous and peritoneal mast cells were 91 +/- 5 and 44 +/- 10%, respectively. In the control group, they were 11 +/- 5 and 7 +/- 2%, respectively. The histamine release rates from sensitized and control rat peritoneal mast cells were 13 +/- 2 and 3.0 +/- 0.4%, respectively. The passive cutaneous anaphylaxis (PCA) test demonstrated that the antibody titers in sensitized serum were 16-32.

[Establishment of epithelial cell line TBC-27 from a human bladder transitional cell carcinoma and its biological characteristics].

The cell line TBC-27, from a human bladder transitional cell carcinoma Grade II, was established and maintained for 8 months through 27 passages. TBC-27 cells have the epithelioid cell morphology during culture. The population doubling time of the 14th generation cells was 27 hours by growth curve. The chromosome karyotype of the 12th generation cells consisted mainly of hypodiploid (mode 40-45). Ultrastructurally, there was a great deal of freeribosome and vesicle in the cytoplasm and a great number of atypical microvilli on the surface of the 21st generation cells. The heterotransplantation test revealed that the xenografted tumor in the immunosuppressed suckling rats was similar to the initial bladder carcinoma in histology.

[Cultivation of urologic and male genital tumor cells].

Thirty specimens from 30 patients with urologic and male genital system tumors were cultured. Eighteen (60%) showed initial cell growth. Four (13%) were successful in primary culture. In a renal cell carcinoma and a bladder transitional cell carcinoma, the subcultures were successful after the primary culture. In eight months, having transferred the former for twelve generations and the latter for twenty-seven generations, these two cell lines are established. The morphological characters of the cultured cells, eradication of fibroblasts, conditions of culture and prevention of contamination are discussed.