Objective To investigate the effect of local injection of rat adipose-derived mesenchymal stem cells (ADSCs)on the phenotype of macrophages in skin wound.Methods The cryopreserved primary SD rat ADSCs were resuscitated and then sub-cultured.ADSCs of the third generation were used in the experiments.Thirty six SD rats were divided into ADSCs group (n =18) and control group (n =18) by random numbers table method.The full thickness skin wounds were established on bothsides of the spine.After the model establishment 0.2 ml ADSCs suspension labeled by live cell stain Chloromethylbenzamido derivatives of 1,l'-dioctadecyl-3,3,3',3'-tetramethylindo-carbocyanine perchlorate (CM-Dil) with the concentration of 5 × 106/ml was subcutaneously annularly injected in the skin wound of SD rats in ADSCs group.The SD rats in control group were given 0.2 ml serum-free Dulbecco modified Eagle medium (DMEM).On 3,7,and 14 days after injury,six rats were selected from each group to measure the wound area and healing rate.The healed wound tissues were harvested to observe the morphology by HE staining.The expressions of interleukin (IL)-10 were detected by immunohistochemistry staining.The double-positive expressions of CD68 and inducible nitric oxide synthase (iNOS) (M1 type macrophages) and of CD68 and arginase-1 (Arg-1) (M2 type macrophages) were detected by immunofluorescent staining.The distribution of CM-Dil-labeled ADSCs in healed wound tissue 14 days after injury was observed by inverted fluorescence microscope.Results (1) At day 3 after injury,the wound areas in two groups were covered with crust and surrounding redness,and the wound healing rates were slightly different;at day 7 after injury,the wound area of ADSCs group was significantly smaller than that of control group,and the healing speed and rate of ADSCs group was significantly higher than that of control group (P ＜ 0.01);at day 14 after injury,the healing rate of ADSCs group was nearly 99％ (P ＜ 0.01),and the healing skin tissue texture of ADSCs group was better than that of control group.(2) At day 3 after injury,there were a large number of inflammatory cells and disorganized collagen fiber in the wound areas of two groups;at day 7 after injury,the inflammatory cells infiltration reduced in ADSCs group compared with control group,and the collagen fiber arrangement in control group was in disorder;at day 14 after injury,the inflammatory cells in both groups obviously decreased,and ADSCs group had more new vessels and more orderly arrangement of collagen fiber than control group.CM Dil labeled ADSCs were seen in the healing wound tissue in ADSCs group.(3) At day 3 after injury,there was little difference in M1 type macrophage distribution in the two groups;ADSCs group had more M2 type macrophage cells than control group significantly (P ＜0.01);the expression of IL 10 in ADSCs group was not high,which did not differ from that of control group;at days 7 and 14 after injury,ADSCs group has fewer M1 type macrophage cells,more M2 type macrophage cells,and higher expressions of IL-10 than control group (P ＜ 0.01).Conclusion The ADSCs trasplantation can promote the change from M1 type to M2 type macrophages,facilitating wound regeneration and healing.

Objective: To investigate the effect of local autologous dermal fibroblasts transplantation on hypertrophic scar formation and wound healing quality in early scar formation. To explore the feasibility of fibroblasts for prevention and treatment of hypertrophic scar. Methods: Dermal fibroblasts were isolated from the dorsal skin tissue of New Zealand white rabbits by mechanical method combined with enzyme digestion. Passage 3 cells were induced to differentiate into osteoblasts and adipocytes. The complete epithelialization time and hypertrophic scar formation after full-thickness skin defect were confirmed by pre-experiment study. In the experiment, 6 rabbits were used, left ear as experimental group and right ear as control group. In the experimental group, the passage 4 dermal fibroblasts labeled with 5-bromodeoxyuridine (5-BrdU) were injected subcutaneously around the wound and hypertrophic scar on 20 d (day 2 after epithelialization) and 30 d (most obvious scar hyperplasia) after surgery. As a control group, physiological saline was injected following the same protocol. On 37 days after surgery, the hypertrophic scar tissue were harvested and assessed by gross view and histological examination. The transplanted cells were detected by immunofluorescence staining, transforming growth factor beta 1 (TGF-β1) and decorin(DCN) mRNA expression were assayed by real-time fluorescence polymerase chain reaction (RT-PCR), and the protein expression of TGF-β1、DCN、Collagen type Ⅰ and type Ⅲ were tested by enzyme linked immunosorbent assay(ELISA). Results: Compared with the control group, the scar in the experimental group was flatter and softer, the color was slightly lighter, and the volume was reduced. The histological results showed that compared with the control group, the number of fibroblasts and inflammatory cells in the superficial dermis was reduced, the proliferation of connective tissue and collagen deposition were reduced, and the basal cells and collagen fibers were arranged in order in the experimental group. The results of RT-PCR showed that TGF-β1 mRNA expression level in the hypertrophic scar tissue reduced significantly and DCN increased significantly in the experimental group, compared with the control group (P<0.01). ELISA results showed that TGF-β1, type Ⅰ and Ⅲ collagen contents in hypertrophic scar tissue were significantly lower than that in the control group, while the DCN expression was significantly increased (P<0.01). Conclusions The rabbit dorsal dermal fibroblasts significantly inhibited the hypertrophic scar in rabbit ears. The mechanism may be related to the up-regulation of DCN expression and down-regulation of TGF-β1 expression, reduction of type Ⅰ and Ⅲ collagen synthesis and extracellular matrix deposition, in the hypertrophic scar tissue.

Objective: To evaluate the outcome of modified rotary-propulsion facial artery perforator flaps for infraorbital defects repair, after facial tumorresection. Methods: Between January 2014 and June 2017, 21 patients with midface tumor were treated, including basal cell carcinoma (n=17) and squamous cell carcinoma (n=4). The size of the tumor ranged from 0.8 cm × 2.0 cm to 2.0 cm× 3.5 cm. The extended resection of the tumor tissue was performed, based on the tumor type. Intraoperative frozen specimen was used to make sure no tumor residual at margin or wound base. According to the location and size, anarterial perforator flap was designed to cover the defect.The donor site was closeddirectly. The flap size ranged from 1.5 cm × 3.0 cm to 3.0 cm × 5.0 cm. The length of flap pedicle was 1.0 cm to 2.5 cm. Results: All flaps completely survived, with satisfying blood flow.The incisions healed well. After 12 to 36 months follow-up, most scars in the donor site were hidden in nasal buccal sulcus and nasolabial folds. There was no lower eyelid ectropion, eyelid and eyeball separation, oral commissural and nasal distortion. The texture and color of the flap was similar to surrounding skin.The overall appearance is satisfactory. Conclusions The modified rotary-propulsion facial artery perforator flaps, to repair soft tissue defects in infraorbital area, is reasonable and useful. It is characterized by limited tissue damage indonor site, flexible rotation of flaps, and hidden incision scars. More importantly, it can reduce the occurrence of lower eyelid ectropion

OBJECTIVETo establish an animal model of sulfur mustard (SM)-induced acute lung injury in rats through different routes and compare the morphological changes in lung tissue and cells.

METHODSOne hundred and thirty-six male rats were selected and randomly divided into 5 groups, namely peritoneal cavity SM group (n=32), trachea SM group (n=32), peritoneal cavity propylene glycol group (n=32), trachea propylene glycol group (n=32), and normal control group (n=8). The rats in peritoneal cavity SM group were injected intraperitoneally with diluted SM (0.1 ml, 8 mg/kg), and the rats in trachea SM group were injected intratracheally with diluted SM (0.1 ml, 2 mg/kg). Once the rats were sacrificed at 6, 24, 48, and 72 h after SM treatment, morphological changes in lung tissue and cells were observed by light and electron microscopy.

RESULTSIn the peritoneal cavity SM group, the epithelial cells of bronchioles maintained intact with increased exudate and bleeding in alveolar cavity and large areas of pulmonary consolidation under the light microscope. In the tracheal SM group, focal ulcer formed in the epithelial cells of bronchioles with increased exudate and bleeding in alveolar cavity, partial pulmonary consolidation, and compensatory emphysema in peripheral alveolar space under the light microscope. The alveolar interval areas were widened obviously in both groups in a time-dependent manner. Under the electron microscope, we observed local loss of cellular membrane in type I alveolar epithelium, broken or lost microvilli in cells of typeⅡalveolar epithelium and fuzzy mitochondrial crista as well as the appearance of ribosome detached from rough endoplasmic reticulum in both two groups. Compared with those in the trachea SM group and the control group, the ratio of the alveolar septum average area to the visual field area in the peritoneal cavity SM group at 6, 24, 48, and 72 h was significantly higher (P<0.05).

CONCLUSIONThe lung tissue injury through the intraperitoneal route is more severe than that through the tracheal route, while focal ulceration of bronchioles epithelial cells appears in the case of tracheal route. The degree of injury increases over time in both groups, and the cellular damage is approximately the same in both groups.

Acute Lung Injury ; chemically induced ; pathology ; Animals ; Disease Models, Animal ; Lung ; pathology ; Male ; Mustard Gas ; toxicity ; Peritoneum ; Pulmonary Alveoli ; pathology ; ultrastructure ; Rats ; Trachea

Toll-like receptors (TLRs) functionally expressed in salivary epithelial cells, but their roles remain elusive. Among TLRs family, TLR3 is activated by dsRNA, a byproduct of viral infection. The aim of this study was to investigate the role of TLR3 in the inflammatory immune responses using HSG cells. Reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR and ELISA were performed to identify expression of TLRs and TLR3-mediated chemokine inductions. The chemotaxis assay of activated T lymphocytes was also performed. Treatment of HSG cells with polyinosinic: polycytidylic acid (poly(I:C)) significantly increased interferon-gamma-inducible protein 10 (IP-10), interferoninducible T-cell alpha chemoattractant (I-TAC), and regulated on activation, normal T-cells expressed and secreted (RANTES) gene expressions in a concentration-dependent manner. Anti-TLR3 antibody blocked the increases of IP-10 and I-TAC genes. Poly(I:C)-induced increases of IP-10 and I-TAC were also confirmed at protein levels from cell lysates, but their release into extracellular medium was detected only in IP-10. We found that the culture media from HSG cells stimulated with poly(I:C) significantly increases T lymphocyte migration. Our results suggest that TLR3 plays an important role in chemokine induction, particularly IP-10, in salivary epithelial cells.
Chemokines ; Chemotaxis ; Culture Media ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; Gene Expression ; Humans ; Lymphocytes ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; Toll-Like Receptors

Objective? To investigate the hope level of elderly patients undergoing total knee arthroplasty (THA) and analyze its influencing factors, so as to provide evidence for improving the hope level and quality of life of patients after THA. Methods? A cluster random sampling method was used to select 308 patients with bilateral THA for the first time who were admitted to two ClassⅢ Grade A general hospitals in Harbin from January 2017 to July 2018. Questionnaire surveys were conducted using Herth Hope Scale, the Second Edition of Beck Depression Scale and Social Support Scale. Results? The total score of hope level in elderly THA patients was (29.96±8.72). Among them, 76.62% (236/308) had moderate to high level of hope and the total score of depression was (19.20±11.75) which was at a moderate level. There were significant differences in the hope level of patients with different gender, place of residence, educational level, marital status and family per capita monthly income (P＜ 0.05). The results of multiple stepwise regression analysis showed that age, sex, family income per capita, depression, negative coping style, subjective support and objective support were the influencing factors of elderly THA patients' hope level (P ＜ 0.05). Conclusions? The THA patients' hope are at a medium level. Medical staff should pay attention to the relevant factors affecting the level of hope, adopt targeted nursing intervention measures, give psychological guidance to patients, provide social support, and further improve the level of hope of the THA patients.

Objective To observe the role of dried tangerine peel in Yigong Powder improves iron metabolism and promotes red blood cell generation in anemia of chronic disease (ACD).Methods With a two-by-two factorial design,the Yigong Powder was divided into dried tangerine peel and Chenpi absent Decoction. According to the random number table method,32 zymosan-induced generalized inflammation (ZIGI) mice were randomly divided into the model group,the dried tangerine peel group,the Chenpi absent Decoction group,and the Yigong Powder group. The dried tangerine peel group,Chenpi absent Decoction group and the Yigong Powder group were given dried tangerine peel(3.083 g/kg),Chenpi absent Decoction(12.33g/kg),and Yigong Powder(15.413g/kg)by gavage to the corresponding group of mice. The model group was given an equal amount of physiological saline by gavage,and treated continuously for 7 days. After the completion of administration,the body weight of each group of mice was recorded. The hemoglobin content of each group of mice was detected using a fully automatic cell counter,the serum iron content was detected using colorimetry,the serum ferritin content was detected using enzyme-linked immunosorbent assay (ELISA),and the spleen index was calculated. The liver tissue inflammatory factors interleukin-1β (IL-1β),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),interferon-γ (IFN-γ),interleukin-4 (IL-4),and interleukin-10 (IL-10) levels were detected using Luminex method. The mRNA expressions of liver tissue hepcidin gene (HAMP) and membrane iron transporter ( Fpn) were detected using real-time fluorescence PCR method. Results Dried tangerine peel and Chenpi absent Decoction both showed interactive effects in regulating hemoglobin,serum iron,serum ferritin content,improving spleen index,and regulating the mRNA expressions of HAMP,Fpn,as well as IL-1β and IFN-γ (P<0.05). Compared with the model group,dried tangerine peel significantly increased hemoglobin,serum iron content,and Fpn mRNA expression in ZIGI model mice,while decreasing ferritin content,spleen index,HAMP mRNA expression,and the levels of IL-1β,IL-6,TNF-α,and IFN-γ (P<0.05). Chenpi absent Decoction significantly increased serum iron content and Fpn mRNA expression in ZIGI model mice,while reducing spleen index,ferritin content,HAMP mRNA expression,and the levels of IL-1β and IFN-γ、IL-4 (P<0.05). Conclusion The effects of dried tangerine peel on inflammatory factors (IL-6 and TNF-α) and Fpn may play a key role in the improvement effects of Yigong Powder on ACD and iron metabolism.