Objective:To explore the efficacy of a flat ground exoskeleton robot in improving the walking ability of stroke survivors.Methods:Fifty-eight stroke survivors with mobility difficulties were randomly divided into a robot group ( n=29) and a control group ( n=29). In addition to routine rehabilitation, the control group received conventional walking training, while the robot group underwent exoskeleton robot-assisted gait training. The 30-minute training sessions were held twice a day, 5 days per week for 5 weeks. Before as well as after 2 and 4 weeks of treatment, everyone′s walking ability was tested using the 6-minute walk test (6MWT) and functional ambulation scale (FAC). General lower limb motor function was quantified using the Fugl-Meyer Lower Extremity assessment (FMA-LE). Moreover, gait analysis was conducted before and after 4 weeks of treatment. Results:After 2 and 4 weeks of treatment, the average 6MWT times of both groups were significantly better than before the treatment, with the improvement of the robot group significantly greater than that of the control group after 2 weeks. After 2 and 4 weeks the average FMA-LE and FAC scores of both groups had improved significantly compared with before treatment. After 4 weeks the stride frequency and gait cycle of both groups had improved significantly.Conclusions:Exoskeleton robot-assisted gait training can improve walking ability and lower limb motor function of stroke survivors about as well as conventional walking training.

Objective To analyze the recurrence of breast cancer without use of chest wall bolus during adjuvant intensity modulated radiotherapy after modified radical mastectomy, so as to investigate the necessity of bolus use. Methods A total of 218 patients undergoing adjuvant intensity modulated radiotherapy after modified radical mastectomy during the period from January 2013 to June 2019 were enrolled as the study subjects. The chest wall bolus was not used during the whole period of radiotherapy after modified radical mastectomy, and the recurrence of breast cancer in the chest wall was analyzed after radiotherapy. Results The post-surgical follow-up through outpatient records, inpatients records, local residents’ health system and telephone was performed until June 31, 2021. The proportion of follow-up was 100%, and the mean follow-up period was 48.9 months. There were three cases with breast cancer recurrence in the chest wall, including one case with recurrence in the chest wall alone and two cases with recurrence in the chest wall and regional lymph nodes, and the overall recurrence of breast cancer was 1.4% in the chest wall. Among the 3 cases with breast cancer recurrence in the chest wall, there were two cases with N3 stage and positive for HER2, and one triple-negative breast cancer case, and all three cases developed distal metastases upon local recurrence. Among 218 study subjects, there were 5 cases with grade Ⅰ radioactive skin reaction, 3 cases with grade Ⅱ radioactive skin reaction, and no grade Ⅲ or Ⅳ radioactive skin reaction occurred. In addition, no grade Ⅲ or Ⅳ acute radioactive injury was seen in the chest wall skin among the 218 study subjects. Conclusion No use of chest wall bolus may be considered during adjuvant intensity modulated radiotherapy after modified radical mastectomy in presence of systemic therapy if tumor invasion into skin is not observed prior to therapy.

Objective:To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods:The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang′an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher′s exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results:The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups ( P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar ( F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining ( t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue ( Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining ( t=4.569, 15.519, P<0.01). Conclusions:Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

Objective:To explore the effects of Freund′s complete adjuvant on autophagy protein expression in rat tuberculous wound model.Methods:The experimental research method was used. In the first batch, twelve 6-week-old male Sprague-Dawley (SD) rats were sensitized by subcutaneous injection of Freund′s complete adjuvant into the hips. Three weeks later, the rats were infected with attenuated Bacille Calmette-Guérin (BCG) subcutaneously on both sides of the back spine. After establishing the tuberculosis wound rat model, according to the random number table (the same grouping method below), the rats were divided into 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group, with 3 rats in each group, with continuous normal feeding to the corresponding days after infection. In the second batch, twenty-three 6-week-old male SD rats were divided into blank control group ( n=3, normal feeding without any treatment), BCG alone group ( n=5), BCG+ rapamycin group ( n=6), BCG+ 3-methyladenine group ( n=6), and BCG+ starvation group ( n=3). The last 4 groups of rats were sensitized as before, and infected as before 1 week later. Rats in BCG alone group were fed normally without any treatment. Rats in BCG+ rapamycin group or BCG+ 3-methyladenine group were intraperitoneally injected with rapamycin or 3-methyladenine once every other day and fed normally. Rats in BCG+ starvation group were fasted for 48 hours after infection and then fed normally. All the rats in the first batch of 4 groups were sacrificed on the corresponding days after infection, and the tissue where the buttocks were injected with Freund′s complete adjuvant was harvested; the tissue of rats in the second batch of BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were harvested the same as before 7 days after infection, and all the rats in blank control group were taken the same tissue at the same time point. Hematoxylin-eosin staining was performed to observe the structure and morphology of cells in the tissue harvested; immunohistochemistry was used to observe the protein expressions of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B) in the tissue harvested. Data were statistically analyzed with Kruskal-Wallis test and Bonferroni correction. Results:Inflammatory cell infiltration was observed in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, granuloma formation was seen in 15 d infection group, part of tissue cell necrosis was seen in 32 d infection group and 43 d infection group, and cell necrosis in 43 d infection group was worse than that in 32 d infection group. Seven days after infection, inflammatory cell infiltration was seen in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group, while regular arrangement of cells and no inflammatory cell infiltration were observed in blank control group. There were no statistically significant differences in the protein expressions of Beclin-1 or LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in 8 d infection group, 15 d infection group, 32 d infection group, and 43 d infection group ( H=1.923, 5.821, P>0.05). Seven days after infection, the protein expressions of Beclin-1 and LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in blank control group, BCG alone group, BCG+ rapamycin group, BCG+ 3-methyladenine group, and BCG+ starvation group were respectively 0.325% (0.250%, 0.360%), 3.225% (1.340%, 3.987%), 4.823% (2.630%, 6.559%), 4.216% (1.790%, 5.969%), 1.765% (0.865%, 2.649%), and 0.301% (0.264%, 0.516%), 2.865% (1.455%, 5.768%), 1.033% (0.398%, 1.873%), 1.168% (0.429%, 1.907%), 0.655% (0.283%, 1.652%). The protein expression of Beclin-1 in the tissue of rats where the Freund′s complete adjuvant was injected in BCG+ rapamycin group was significantly higher than that of blank control group ( Z=4.796, P<0.05). The protein expression of LC3B in the tissue of rats where the Freund′s complete adjuvant was injected in BCG alone group was significantly higher than that of blank control group ( Z=4.953, P<0.05). Conclusions:Freund′s complete adjuvant can enhance the expression levels of local tissue autophagy-related proteins Beclin-1 and LC3B in rat tuberculous wound model.

Objective:To evaluate the reliability of a rat tuberculous wound model established by injecting Bacillus Calmette- Guérin (BCG). Methods:The experimental research was conducted. According to the random number table, fifteen 6-week-old male Sprague-Dawley rats were divided into normal control group and infection group, with 3 rats in normal control group and 12 rats in infection group. Rats in infection group were injected with Freund's complete adjuvant, 3 weeks later, they were injected subcutaneously with BCG bacterial solution to establish a model of tuberculous wounds in rats; rats in normal control group did not receive any treatment. On the 8th, 15th, 32nd, and 43rd day of infection, the skin condition at the injection sites of the rats in infection group was observed roughly. Skin tissue at the injection sites of 3 rats in infection group at each corresponding time point stated above and skin tissue at the corresponding sites of the rats in normal control group were stained with hematoxylin-eosin to observe the cell arrangement, necrosis and inflammation. On 43rd day of infection, acid-fast staining was performed on the skin tissue at the injection sites of the rats in infection group to observe the distribution of bacteria.Results:On the 8th, 15th, 32nd, and 43rd day of infection, tuberculous wound lesions were gradually developed at the skin tissues at the injection sites of the rats in infection group. The cells of the diseased tissue of the rats in infection group arranged disorderly or concentrically, and the number of granulomas and necrotic cells gradually increased, while the skin tissue cells in the corresponding parts of the rats in normal control group arranged regularly with no inflammatory cell infiltration. On the 43rd day of infection, a large number of rod-shaped bacteria were observed in the skin tissue at the injection sites of the rats in infection group.Conclusions:The rat tuberculous wound model established using BCG is stable and reliable, which can meet the experimental requirements.

Foamy macrophages (FM), also known as foam-like macrophages, refer to lipid-laden monocytes or macrophages. FM are a kind of inflammatory cells that are rich in lipid droplets in cytoplasm. In the diseases caused by Mycobacterium tuberculosis (Mtb), such as granuloma and tuberculous wounds, FM can not only inhibit the immune response, but also affect the prognosis and outcomes. The formation mechanisms of FM caused by Mtb infection have some specificity, which may be an important factor for its long-term survival in cells and influences on disease prognosis and outcomes. Therefore, studying the mechanisms of Mtb-mediated formation of FM is conductive to further reveal the pathological evolution of diseases and provide new ideas for further precise treatment. This article reviewed the mechanisms of Mtb-mediated formation of FM in recent years.

Chronic refractory wound refers to the wound with unclear etiology, multiple and complex injury factors, slow healing, and no obvious tendency of healing after treatment for 4 weeks. The formation and evolution process of chronic refractory wounds are very complex, involving re-epithelialization of wound tissue, cell proliferation, tissue remodeling, and angiogenesis and lymphangiogenesis. The abnormal expression of long non-coding RNA may be involved in the formation of chronic refractory wounds, but the specific pathogenesis and related molecular biological changes are still controversial. In this paper, we reviewed the process and role of long non-coding RNA in regulating keratinocyte differentiation, fibroblast proliferation, and regeneration of vascular and lymphatic endothelial cell in chronic refractory wounds.

Objective To study the temporal distribution of macrophage and its phenotype markers in fibrous capsules around silicone implants.Methods Thirty rats were randomly divided into five groups:days 1,3,7,14 and 35.Silicone prostheses (10 ml) were implanted subcutaneously into backs of rats.On each indicated day,the tissue specimens were collected,fixed in 4％ paraformaldehyde for 24 hours and embedded in paraffin.Immunofluorescence was used to detect temporal distribution of M1/M2 macrophages.Results The number of CD68+ macrophages at day 1 (65.8±12.9) was smaller than that at day 3 (102.8±14.5,P＜0.05) and day 7 (116.8±14.2,P＜0.05);and the number of CD68+ macrophages at day 7 was larger than that at day 14 (56.8±12.9,P＜0.05) and day 35 (21.40±6.35,P＜0.05);the proportion of iNOS+ CD68+ M1 cells at day 1 and day 3 was 0.48±0.13,0.60±0.13,respectively,and they were higher than that at day 7 (0.21±0.03,P＜0.05),day 14 (0.21±0.03,P＜0.05) and day 35 (0.17±0.04,P＜0.05);the proportions of CD206+ CD68+ M2 cells at day 1,day 3,day 7,day 14,day 35 were 0.70±0.06,0.60±0.07,0.70±0.08,0.67±0.02 and 0.60±0.06,respectively.Conclusions After the implantation of silicone prostheses,M1 cells increase in early stages and M2 cells maintain in high level throughout the experiment period.

Objective: To investigate the functional recovery and cosmetic effects of pre-expanded pedicled deltopectoral flap. Methods: From January 2008 to December 2018, 42 patients with 56 pre-expanded pedicled deltopectoral flaps from Xijing Hospital of Air Force Military Medical University were followed up at least 6 months. 18 of them were male, the remaining were female. And the average age was (24.7±7.3) years. Then the indicators were tested and evaluated. Tubes with different temperatures were used for temperature sensation test. The flaps were stabbed using a 27 G blunt needle to test algesthesia. Tactile threshold was measured by Semmes-Weinstein monofilament. Two-point discrimination was measured by Disk-Criminator. Elasticity of skin flaps was measured by CK-MC®960. Colors of skin flaps were analyzed by ANTERA®3D system, including L^*a^*b^*, melanin and hemoglobin content. And the postoperative scars were evaluated by the Patient and Observer Scar Assessment Scale (POSAS). The flap retraction rate [(flap area immediately after operation-flap area at the follow-up time)/flap area immediately after operation]was calculated. The satisfaction of patients, doctors and third parties was investigated as well. Statistical analysis of data was performed with SPSS 23.0, satisfaction rate was expressed as percentages. Scar scores were compared by Wilcoxon rank sum test. The values of elasticity, color L^*a^*b^*, melanin and hemoglobin between skin flaps and normal skin were analyzed by paired t test. The difference was statistically significant at P<0.05. Results: The scores of pain, heat and cold sensation were (4.61±0.78), (3.48±0.90) and (4.39±0.81), the tactile threshold was (0.26±0.23) g/mm², and the distance of two-point discrimination was (10.09±4.79) mm. There was no significant difference in elasticity, color b^*and melanin content between skin flap and normal skin of face and neck (P>0.05). The color of flap L^*, a^*and hemoglobin content of flaps were significantly different from those of normal skin from face and neck (P<0.05). The result of self-assessment of patients (POSAS) showed that there were significant differences (P<0.05) in pain, itch and stiffness between facial and chest donor scar. The result of observer evaluation (POSAS) showed that the scar score for all items around facial flap was better than that of chest donor area, the difference was statistically significant (P<0.05), and the retraction rate of flaps was (10.44 ±3.36)%. The satisfaction rates of doctors, patients and the third party were 92.86% (39/42), 71.43% (30/42) and 61.90% (26/42), respectively. Conclusions Excellent functional recovery and reliable cosmetic effect were observed in the facial-cervical scar repair with pre-expanded pedicled deltopectoral flap. Transfer of pre-expanded pedicled deltopectoral flap is an effective method for the treatment of facial-cervical scars.

Objective To explore the anatomic basis of the thinning of the free posterior tibial artery perforator flaps and the clinical effect of repairing wound on hand or foot due to trauma.Methods From November,2016 to December,2017,10 cases of lower extremity cadaver specimens perfused with red ralex were dissected,which were perfused through the amputated femoral artery.Five of them were left and the rest were right.All cases were males.The number,diameter,branches and distribution of the perforator was observed.From September,2012 to September,2017,there were 13 cases of clinical application,which were 5 cases of hand wound and 8 cases of foot wound.The size of the wound was 3.0 cm × 2.0 cm to 6.0 cm × 4.0 cm,and the flap area was 3.5 cm × 2.2 cm to 6.5 cm × 4.5 cm.The repairing procedure was suitable for the wound associated with tendon,bone,joint capsule exposure.Results The number of posterior tibial artery perforating branches that more than 0.50 mm in diameter was 4 to 6,and the mean diameter was (0.87±0.26) mm.The perforating branch penetrated into the fat layer and was divided into 3 layers of vascular network:deep fat vascular network,superficial fat vascular network and subdermal vascular network.The perforating branch was located according to the positional relationship from deep to shallow,and vessel diameter become smaller step by step.The perforating branch trunk gave off branches to the deep vascular network,and the superficial vascular network had the same origin or shared with the deep blood vessels.The subdermal vascular network issued from the superficial vascular network or directly from the perforating branch trunk.There was no or few communicating branch between the deep vascular network and superficial ones,besides the vessel pedicle.So trimming deep fat layer will not affect the blood supply of superficial vascular network and neither will affect the flap blood supply.Most of the deep fat tissue was trimmed in 13 cases.The superficial and subdermal fat vascular network was preserved,and the same to the trunk and branches of the pedicle.All the flaps survived.Of which,1 appeared arterial crisis on the 2nd day after operation,and relieved by the local injection of papaverine.There was 1 case of venous crisis on the 3rd day,and improved by stitches,local release of congestion.Followed-up time was ranged from 2 to 12 months.All flaps were soft with good blood supply and good appearance,and did not need a second thinning surgery.Flaps restored the protective feeling 6 months later.Conclusion The microdissection of perforator flap of posterior tibial artery provides a theoretical basis for the perforator flap thinning,and the thinning of perforator flap is a good method to repair the appearance and function of the wound after foot and hand injuries.