Objective: To explore the effects of scar excision combined with negative-pressure on repair of hypertrophic scar in burn children. Methods: From October 2010 to August 2016, 25 children with hypertrophic scar after deep burn were hospitalized, with scar course ranging from 3 months to 11 years and scar area ranging from 35 to 427 [83(51, 98)]cm². A total of 35 scars of 25 children were located in trunk (11 scars), upper limb (11 scars), and lower limb (13 scars). All children received scar excision operation and negative-pressure treatment (negative-pressure value ranged from -40 to -20 kPa), among which 6 cases received scar excision operation and negative-pressure treatment for two times for further removal of scars. After scar excision, electronic spring scale was used to measure the tension of the incision. The tension value of children ranged from 3.43 to 23.84 [7.16 (5.59, 9.12)] N, and then the incision was closed with appropriate suture according to the value of the tension. The incision with smaller tension was firstly opened on post operation day (POD) 8. After removing the suture, negative-pressure was conducted to POD 14. The incision with larger tension was firstly opened on POD 12. After removing the suture, biological semi-membrane was used to reduce tension to POD 16. All healed incisions were performed with anti-scar treatment for 1 year and relaxation and fixation for 3 months. General condition of the incision was observed after operation. The reduction percentage of scar area was calculated half-year after operation. The Patient and Observer Scar Assessment Scale was used to record the overall score of scar and scar score of trunk, upper limb, and lower limb before operation and half-year after operation. Data were processed with paired t test and Wilcoxon rank sum test. Results: After removing the suture, all incisions of children healed well without redness, effusion, and rupture. Half-year after operation, the appearance and deformity of incision were obviously improved, and the symptoms including pruritus and pain were basically relieved. Half-year after operation, the scar area of children ranged from 0 to 174 [21(9, 47)]cm^2, which was significantly decreased as compared with that before operation (Z=-5.16, P<0.05). The reduction percentage of scar area ranged from 36% to 100% [(73±19)%]. Half-year after operation, the overall score of scar and scar score of trunk, upper limb, and lower limb of children were obviously decreased as compared with those before operation (with t values from 6.42 to 17.37, P values below 0.05). Conclusions Scar excision combined with negative-pressure treatment has a good clinical effect on repair of hypertrophic scar in burn children, which is suitable for clinical application.

Objective: To establish a method for repairing extremities with extensively deep burn using large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin and observe its effect. Methods: Medical records of two male patients with extremely extensive deep burn admitted to our hospital from May to November in 2018 were retrospectively analyzed. Two patients aged 44 and 25 years respectively, with total burn area of 90% and 97% total body surface area (TBSA) and full-thickness burn area of 85% and 70% TBSA, respectively. Preoperatively, the surgical area on the extremities was calculated to estimate the necessary amount of allogeneic scalp and Meek miniature skin. The large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin was prepared according to the methods described as follows. Thin medium-thickness fresh scalps with 3% TBSA and 0.30-0.35 mm in depth were harvested from each donor and spliced into a large piece with epidermis upward by spraying Meek glue. Then the spliced scalp was punched after covered with a single-layer gauze. Autologous microskin was transported onto the dermis of fresh large piece of allogeneic scalp by traditional floating method. Bilateral extremities with full-thickness burn of two patients were selected for self-control. The left upper extremity was denoted as treatment group while the right upper extremity was denoted as control group in Patient 1. The right lower extremity was denoted as treatment group while the left lower extremity was denoted as control group in Patient 2. Wounds in the treatment group were treated with fresh large piece of allogeneic scalp spliced by Meek glue and autologous microskin with expansion ratio of 1∶15 after escharectomy, while wounds in control group received grafting of Meek miniature skin with expansion ratio of 1∶6 and or 1∶9 after escharectomy. The donors of allogeneic scalp were 32 males who were the relatives or friends of the patients, aged 21-50 years, with scalp area of (548±48) cm². The healing conditions of donor sites of scalp were observed on post operation day 10, and were followed up within 3 months after operation to observe whether forming alopecia and hypertrophic scar or not. Wound healing condition was evaluated during follow-up in post operation week (POW) 2-5 and 4 months after operation. Wound coverage rates were calculated in both treatment and control groups in POW 2, 3, 4, and 5. Results: The donor sites of all allogeneic scalp of donors healed completely on post operation day 10. There was no alopecia or hypertrophic scar within 3 months after operation for follow-up. In POW 2, allogeneic scalp grafts basically survived in treatment group without obvious exudation, and most of the Meek miniature skin survived in control group with obvious exudation. Part of allogeneic scalp grafts dissolved and detached in treatment group in POW 3, and the surviving grafts scabbed. The eschar detached and new epithelium was observed in treatment group in POW 4 and 5. In POW 3-5, surviving Meek miniature skin in control group creeped and was incorporated, and the wounds shrank. Hypertrophic scar was observed in both treatment and control groups 4 months after operation, without obvious difference in scar as a whole. The wound coverage rates were respectively 84%-98% and 76%-92% in treatment group of two patients in POW 2-5, close to or higher than those of control group (35%-97% and 28%-81%, respectively). Conclusions The study establishes a novel method for splicing fresh allogeneic scalps into a large piece as the covering of microskin, which has good effect for repairing extensively deep burn wounds. Considering that allogeneic skin is scarce, this method may be a new option in clinical treatment for extensively deep burn patients.

ObjectiveTo explore the mechanism of cucurbitacin B （CuB） in inhibiting cell proliferation and glycolysis. MethodCell counting kit-8 （CCK-8） was applied to investigate the effect of different concentrations of CuB （0， 40， 80， 120， 160， 200， 400， and 800 nmol·L^-1） on the proliferation of HuCCT1 cells. The effect of different concentrations of CuB （50， 100， and 200 nmol·L^-1） on the colony formation ability of HuCCT1 cells was detected by plate cloning assay. The effect of different concentrations of CuB （50， 100， 200 nmol·L^-1） on the HuCCT1 cell cycle was analyzed by flow cytometry. Visible spectrophotometry was employed to detect the activity of key glycolytic enzymes hexokinase （HK） and pyruvate kinase （PK）） and changes in glucose consumption， lactate production， and adenosine triphosphate （ATP） production in HuCCT1 cells after administration of different concentrations of CuB （50， 100， 200 nmol·L^-1）. Western blotting was used to assay the effect of CuB on the expression of cell cycle-related proteins， proliferation-related proteins， key glycolytic proteins， and Akt/mammalian target of rapamycin （mTOR） pathway-related proteins. ResultAs compared with the blank group， CuB at dose of 160-800 nmol·L^-1 after 24 h administration and CuB at dose of 80-800 nmol·L^-1 after 48 h administration inhibited the proliferation of HuCCT1 cells in a time- and dose-dependent manner （P<0.05， P<0.01）， and the median inhibitory concentration was 200 nmol·L^-1 48 h after administration. CuB can restrain the colony formation ability of HuCCT1 cells in a dose-dependent manner （P<0.01）， and block HuCCT1 cell cycle in G₂ phase （P<0.05， P<0.01）. CuB （100 and 200 nmol·L^-1） can suppress the activities of HK and PK and reduce cell glucose consumption and production of lactate and ATP （P<0.05， P<0.01）. Western blot results showed that CuB （100 and 200 nmol·L^-1） can inhibit the protein levels of cycle-related protein Cyclin B₁， proliferating cell nuclear antigen （PCNA）， HK1， HK2， PKM1， PKM2， phosphorylated Akt （p-Akt）， phosphorylated mTOR （p-mTOR）， and phosphorylated ribosomal protein S6 （p-RPS6） （P<0.05， P<0.01）. ConclusionCuB can inhibit aerobic glycolysis in HuCCT1 cells via the Akt/mTOR pathway， thereby affecting cell proliferation.

ObjectiveTo investigate the effect and mechanism of osthole on the proliferation and apoptosis in human intrahepatic cholangiocarcinoma HuCCT1 cells. MethodThe effect of 10， 20， 40， 80， and 120 μmol·L^-1 osthole on the proliferation of HuCCT1 cells was detected by the cell counting kit-8 （CCK-8）. A blank group， and low-， medium-， and high-dose osthole groups （16， 32， and 64 μmol·L^-1） were set up. The effect of osthole on cell clone formation rate was detected by colony formation assay. The effect of osthole on cell cycle and apoptosis was detected by flow cytometry. The effect of osthole on cell apoptotic morphology was detected by Hoechst 33342 fluorescent staining. The effect of osthole on cell cycle protein cyclin B₁， proliferating cell nuclear antigen （PCNA）， cysteine-aspartic acid protease （Caspase）-9， Caspase-3， cleaved Caspase-9， cleaved Caspase-3， cleaved poly（ADP-ribose） polymerase （cleaved PARP）， B-cell lymphoma-2 （Bcl-2）， phosphorylated protein kinase B （p-Akt）， phosphorylated mammalian target of rapamycin （p-mTOR）， and phosphorylated ribosomal protein S6 （p-RPS6） was detected by Western blot. ResultThe cell viability in the osthole group（40，80，120 μmol·L^-1） decreased （P<0.05，P<0.01）， with the half maximal inhibitory concentration （IC₅₀） of 63.8 μmol·L^-1 as compared with that in the blank group. Compared with the blank group， the osthole groups（32，64 μmol·L^-1）showed reduced clone formation rate （P<0.01）， increased number of cells in the G₂ phase （P<0.05，P<0.01）， decreased number of cells， increased pyknosis and fragmentation， increased apoptosis rate （P<0.05，P<0.01）， down-regulated expression of cyclin B₁， PCNA， Bcl-2， Caspase-3， Caspase-9， p-Akt， p-mTOR， and p-RPS6 （P<0.05，P<0.01）， and up-regulated expression of cleaved Caspase-3， cleaved Caspase-9， and cleaved PARP （P<0.05，P<0.01）. ConclusionOsthole can inhibit the proliferation and promote the apoptosis of HuCCT1 cells， and its mechanism may be related to the Akt/mTOR signaling pathway.