Objective To evaluate the clinical application of 2000 ArthroCare System in knee arthroscopic surgery. Methods 221 cases of knee problems were treated with 2000ArthroCare System. The disorders of the 221 cases diagnosed by the arthroscopy were as follows: 73 cases of osteoarthritis, 49 meniscus tear, 29 degenerative cartilage injury, 11 plica synovitis, 11 Kaschin Beck disease, 8 ACL, 5 osteochondritis dissecans, and 2 TKA brisement. The operative procedures, such as meniscectomy, meniscoplasty, fitting of cartilage and ligament, synovectomy, and release of lateral patellar retinaculum, were done with 2000ArthroCare System and arthroscopy. Results The Lysholm Knee scores were 43.92 preoperatively, 81.96 three months postoperatively, and 92.06 six months postoperatively. Conclusion Knee problems can be effectively released with 2000 ArthroCare system vaporization under arthroscopic guidance. The advantages of this procedure are very limited tissue damage, mild reaction, less blood loss, early rehabilitation, and fine functional recovery.

OBJECTIVETo investigate the rationality of N3 classification and its sub-classification in the 7th UICC/AJCC TNM classification system.

METHODSClinicopathological data of 610 patients with stage N3 advanced gastric cancer who underwent standard D2 and D2+ radical surgery at the Department of Surgical Oncology, The First Hospital of China Medical University, from January 1980 to March 2010 were analyzed retrospectively. Patients were divided into N3a and N3b groups, and clinicopathological characteristics and prognosis were compared between N3a and N3b patients. Overall survival rate was determined using the Kaplan-Meier estimator. The log-rank test was used to identify differences between the survival curves of different groups. In multivariate analysis, Cox proportional hazard model was used to identify independent factors associated with prognosis.

RESULTSAmong 610 patients, 426 were men and 184 were women, 394 were N3a and 216 were N3b, with a mean age of(57±11) years old (range 23 to 83). A total of 19 842 lymph nodes were examined, in which 9 575 nodes were positive, with the metastatic ratio of 48.3%. The 5-year overall survival rate was 20.0%. Univariate analysis of prognostic factors suggested that tumor location (P=0.000), tumor size (P=0.003), Borrmann type (P=0.000), pathologic type (P=0.043), lymphatic vessel invasion (P=0.000), growth pattern (P=0.019), invasion depth (P=0.000), resection extent (P=0.000) and N3 sub-classification (P=0.000) were significantly associated with the prognosis of N3 patients. Further analysis showed that tumor size (P=0.028), invasion depth (P=0.000) and gastric resection extent (P=0.002) were significantly associated with the prognosis of N3a patients, while Borrmann type (P=0.034), lymphatic vessel invasion (P=0.002), invasion depth (P=0.008) and resection extent (P=0.003) were significantly associated with the prognosis of N3b patients. Multivariate analysis revealed that lymphatic vessel invasion (P=0.009), resection extent (P=0.001), invasion depth (P=0.000) and N3 sub-classification (P=0.000) were independent prognostic factors of N3 patients; resection extent (P=0.004) and invasion depth (P=0.001) were independent prognostic factors of N3a patients; lymphatic vessel invasion (P=0.006) and invasion depth (P=0.009) were independent prognostic factors of N3b patients. Comparison of 5-year survival rate revealed that there was significant difference between T2-4N3a and T2-4N3b patients (P=0.000), while there was no significant difference between T2N3a and T2N3b patients (P=0.140). On the contrary, there were significant differences between T3N3a and T3N3b patients, T4aN3a and T4aN3b patients, T4bN3a and T4bN3b patients, respectively (all P<0.05). Further comparison demonstrated that there were significant differences between T4aN3a and T4bN3a patients, T4aN3b and T4bN3b patients, respectively (P=0.000, P=0.041). Besides, there were no significant differences in 5-year survival rate between T2N3 (at present, staged as III(A), T3N3a (III(B) and T4aN3a (III(C) patients(P=0.506), and T3N3b (III(B), T4aN3b (III(C) and T4bN3a (III(C) patients(P=0.283), respectively.

CONCLUSIONSN3 sub-classification should be included in the final TNM classification system. It is suggested that T2N3, T3N3a and T4aN3a may be categorized into III(A stage, T3N3b, T4aN3b and T4bN3a may be categorized into III(B stage,T4bN3b may be categorized into III(C stage or IIII( stage.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrectomy ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Stomach Neoplasms ; classification ; Surgical Oncology ; Survival Rate ; Young Adult

Objective To investigate the influence of necroptosis pathways in microglia activation and inflammatory factor levels in cerebral ischemia/reperfusion (I/R) injury in rats and their significances.Methods A modified suture method was used to establish the models of middle cerebral artery I/R in rats.(1) SD rats according to the random number table were divided into cerebral I/R 6 h group,cerebral I/R 12 h group,cerebral I/R 24 h group,and cerebral I/R 72 h group (n=6);the expression of ionized calcium binding adaptor molecule-1 (iba-1) was detected by immunohistochemical staining,and time points enjoyed obvious iba-1 expression were selected according to the experimental results.(2) SD rats were randomly divided into sham operation group,cerebral I/R group,80 nmol necrostatin-1 (Nec-1) intervention group and 160 nmol Nec-1 intervention group (n=6),and the Nec-1 intervention was given 2 h after ischemia;Longa method was used to evaluate the neurological function scores and TTC method was used to detect the infarct volume;and the appropriate dosages of Nec-1 were selected according to these results.(3) SD rats were randomly divided into sham operation group,cerebral I/R group,solvent group and Nec-1 intervention group (n=6),and Nec-1 or DMSO intervention was given 2 h after ischemia.HE staining was used to observe the survival and proliferation ofmicroglias around the infarction tissues;immunohistochemical staining was used to detect the iba-1 expression surrounding the infarction tissues;immunohistochemistry and Western blotting were employed to observe the cytokine tumor necrosis factor (TNF)-α and interleukin (IL)-1β and glia-derived neurotrophic factor (GDNF) expressions.Results (1) The iba-1 expression at cerebral I/R 24 h group was significantly increased as compared with that in other groups (P＜0.05).(2) As compared with those in the cerebral I/R group,the neurological function scores and infarct volume were significantly decreased in the 80 nmol Nec-1 intervention group and 160 nmol Nec-1 intervention group (P＜0.05);more obvious changes were noted in the 160 nmol Nec-1 intervention group as compared with those in the 80 nmol Nec-1 intervention group,with significant difference (P＜0.05).(3) HE staining showed peri-infarct tissue inflammatory cell infiltration,reduced neuron number,cell body shrinkage and nuclear pyknosis in the cerebral I/R group,while these changes were less obvious in the Nec-1 intervention group;as compared with cerebral I/R group,the Nec-1 intervention group had significantly decreased expressions of iba-1,TNF-α,IL-1 β and GDNF (P＜0.05).Conclusion Necroptosis pathway involves in the activation ofmicroglias after I/R injury in rat brains;Nec-1 inhibits the activation of microglia,and reduces the neuronal damage by regulating inflammatory cytokines.

Objective: To study the clinical effect of nano-fat mixed granule fat transplantation in the treatment of cicatricial facial depression and atrophy, and to explore the related experimental mechanism. Methods: (1) From January 2012 to April 2018, 105 patients conforming to the inclusion criteria, with cicatricial facial depression and atrophy deformity who needed facial fat transplantation, were admitted to our unit. Their medical records were analyzed retrospectively. According to the patients′ wishes, 54 patients (12 males and 42 females) aged 10-59 years received traditional transplantation of pure autologous granule fat collected from abdomen/thigh and were included in simple transplantation group, while the other 51 patients (14 males and 37 females) aged 7-63 years received transplantation of autologous nano-fat mixed granule fat collected from abdomen/thigh and were included in mixed transplantation group. The treatment satisfaction of patients 3 and 6 months after operation was assessed by the facial fullness, symmetry, scar, and complications using self-made scales and photo data before and after operation. Six months after operation, the patients were assessed whether they needed to undergo a second operation, and the rate of second operation was calculated. During the second operation, the fat of patients transplanted in the first operation was collected, and the morphology of adipocytes and microangiogenesis was observed under a scanning electron microscope. (2) Adipose-derived stem cells (ADSCs) were isolated and cultured from abdominal fat of a 4-week-old male Sprague-Dawley (SD) rat. The 5th passage of cells were selected to observe cell morphology after cultured for 14 days, observe expression of vimentin and cytokeratin-18 by immunofluorescence method, identify osteogenic and adipogenic differentiation, and detect rates of CD29 and CD44 positive cells by flow cytometer (n=3). Eighteen 4-week-old male SD rats were divided into ADSCs transplantation group, simple scar group, and blank control group according to the random number table, with 6 rats in each group. Rats in ADSCs transplantation group and simple scar group were subcutaneously injected with 1 mL bleomycin which was dissolved in phosphate buffered saline (PBS) with a mass concentration of 1 mg/mL at the back to establish scar models. After 3 hours, rats in ADSCs transplantation group were injected with 1×10⁶ ADSCs suspended in 0.1 mL PBS at the same injection site, while rats in simple scar group were injected with 0.1 mL PBS. Rats in blank control group were injected with the same doses of PBS in the same place at the same two time points mentioned above. After continuous injection for 28 days in each group, the full-thickness skin tissue of the injected area of all rats was collected to observe the collagen fibers by Masson staining and expressions of α-smooth muscle actin (α-SMA) and transforming growth factor β₁ (TGF-β₁) by immunohistochemistry, and the positive cells were counted. Data were processed with Mann-Whitney U test, χ² test, one-way analysis of variance, and least significant difference test. Results: (1) Compared with the preoperative condition, the facial fullness and symmetry of patients in simple transplantation group were better in 3 months after operation, with scar color closer to the surrounding skin, and the filling volume of patients in this group decreased in 6 months after operation as compared with that in 3 months after operation. In mixed transplantation group, the facial fullness and symmetry of patients were better in 3 and 6 months after operation as compared with the preoperative condition, with scar color and texture closer to the surrounding skin, and the filling volume in 6 months after operation was not obviously reduced as compared with that in 3 months after operation. Fat liquefaction and subcutaneous nodule formation occurred respectively in 1 patient in simple transplantation group within 3 months after operation. The treatment satisfaction of patients in mixed transplantation group was significantly higher than that in simple transplantation group in 3 and 6 months after operation (Z=-2.566, -3.084, P<0.05 or P<0.01). Six months after operation, the second operation rate of patients in mixed transplantation group was 7.84% (4/51), which was significantly lower than 22.22% (12/54) in simple transplantation group (χ²=4.199, P<0.05). At the second operation, compared with those of simple transplantation group, the cells of fat transplanted in the first operation of patients in mixed transplantation group were more plump, without collapse or dryness, and the cells were closely arranged, with smaller gap; the tubular and the cord-like microvascular structure on the cell surface were more abundant, and the cell gap was full of network-like microvascular structure that grew into the adipose tissue. (2) The fifth passage of cells isolated and cultured from rat fat grew adherently to the wall, with long fusiform or spindle shape, showing shoal-of-fish-like growth. Vimentin and cytokeratin-18 were highly expressed in the cells. Cells showed osteogenic and adipogenic differentiation ability by induction. The positive expression rates of CD29 and CD44 were higher than 90.00%. The cells were identified as ADSCs. After 28 days of injection, the collagen fibers in the dermis of skin tissue at the injection area of rats in blank control group were finely arranged. In simple scar group, a large amount of collagen was deposited in the dermis of skin tissue at the injection area of rats, the fiber bundles were thick and loosely unevenly arranged, and a large number of inflammatory infiltration and scattered muscle fibers were observed. In ADSCs transplantation group, the collagen fibers in the dermis of skin tissue at the injection area of rats were thicker than those of blank control group, with still neat arrangement, and a small amount of scattered muscle fiber and inflammatory infiltration was observed. After 28 days of injection, the expression of α-SMA in ADSCs transplantation group was mainly in microvessels in the dermis of skin tissue at the injected area of rats, and the number of α-SMA and TGF-β₁ positive cells was (49±12) and (63±10) cells per 20-fold field of view, respectively, which was similar to (35±16) and (44±17) cells per 20-fold field of view of blank control group (P>0.05), all significantly less than (135±13) and (121±23) cells per 20-fold field of view of simple scar group (P<0.05). Conclusions Compared with those of autologous simple granule fat transplantation, autologous nano-fat mixed granule fat transplantation has better filling fullness in the treatment of patients with scar facial depression and atrophy. The filling effect lasts longer, and the improvement of scar texture is more obvious. As showed in the rat scar model experiment, the mechanism may be that ADSCs inhibit the expressions of α-SMA and TGF-β₁, thus inhibiting the formation of scar.

Sepsis is life-threatening multiple organ failure (MOF) caused by the disorder of body's response to infection. Although it is the main reason of death in intensive care patients, the mechanism of sepsis-induced MOF is still unclear. As an essential organelle of cells, mitochondrial is responsible for many funtions in cells such as energy supply and signal regulation. Recent researches showed that mitochondrial dysfunction may be one of the important reasons of MOF. This review describes the pathophysiological functions of mitochondria and the pathological damage mechanisms of MOF in sepsis, it will provide the therapeutic targets or new ideas for clinical treatment of sepsis.

The aim of this study was to construct a recombinant adenovirus expressing extracellular domain gene of human epidermal growth factor receptor variant Ⅲ (EGFRvIII ECD), and to prepare single domain antibody targeting EGFRvIII ECD by immunizing camels and constructing phage display antibody library. Total RNA was extracted from human prostate cancer cell line PC-3 cells and reversely transcribed into cDNA. EGFRvIII ECD gene was amplified using cDNA as template, and ligated into pAdTrack-CMV plasmid vector and transformed into E. coli BJ5183 competent cells containing pAdEasy-1 plasmid for homologous recombination. The recombinant adenovirus expressing EGFRvIII ECD was obtained through transfecting the plasmid into HEK293A cells. The recombinant adenovirus was used to immunize Bactrian camel to construct EGFRvIII ECD specific single domain antibody library. The single domain antibody was obtained by screening the library with EGFRvIII protein and the antibody was expressed, purified and identified. The results showed that recombinant adenovirus expressing EGFRvIII ECD was obtained. The capacity of EGFRvIII specific phage single domain antibody library was 1.4×10⁹. After three rounds of enrichment and screening, thirty-one positive clones binding to EGFRvIII ECD were obtained by phage-ELISA, and the recombinant single domain antibody E14 with highest OD₄₅₀ value was expressed and purified. The recombinant E14 antibody can react with EGFRvIII ECD with high affinity in ELISA assessment. The results indicated that the EGFRvIII specific single domain antibody library with high capacity and diversity was constructed and the single domain antibody with binding activity to EGFRvIII was obtained by screening the library. This study may facilitate the diagnosis and treatment of EGFRvIII targeted malignant tumors in the future.
Adenoviridae/genetics* ; DNA, Complementary ; ErbB Receptors ; Escherichia coli/genetics* ; Genetic Vectors/genetics* ; Humans ; RNA ; Recombinant Proteins/metabolism* ; Single-Domain Antibodies