Tanshinone ⅡA activates PI3K/AKT signaling pathway to inhibit the apoptosis of mice cochlear pericytes induced by high glucose / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate whether tanshinone ⅡA can protect the apoptosis of mice cochlear pericytes induced by high glucose and its specific protective mechanism, so as to provide experimental evidence for the prevention and treatment of diabetic hearing loss. Methods: C57BL/6J male mice were used to prepare type 2 diabetes model, which were divided into normal (NG) group, diabetic (DM) group, diabetic+tanshinone ⅡA (HG+tanshinone ⅡA) group and tanshinone ⅡA group. Each group had 10 animals. Primary cochlear pericytes were divided into NG group, HG group (high glucose 35 mmol/L), HG+tanshinone ⅡA (1, 3, 5 μmol/L) group, HG+Tanshinone ⅡA+LY294002 (PI3K/AKT pathway inhibitor) group, LY294002 group, tanshinone ⅡA group and DMSO group. Auditory brainstem response (ABR) was used to measure hearing threshold. Evans blue was used to detect the permeability of blood labyrinth barrier in each group. TBA methods were used to detect oxidative stress levels in various organs of mice. Morphological changes of stria vascularis were observed by hematoxylin-eosin staining (HE). Evans blue was used to detect the vascular labyrinth barrier permeability in cochlea. The expression of apoptosis protein in stria vascularis pericytes was observed by immunofluorescence. Pericytes apoptosis rate was observed by flow cytometry. DCFH-DA was combined with flow cytometry to detect intracellular ROS content, and Western blot was used to detect the expression of apoptotic proteins (Cleaved-caspase3, Bax), anti-apoptotic proteins (BCL-2) and pathway proteins (PI3K, p-PI3K, AKT, p-AKT). SPSS software was used for statistical analysis. Independent sample t test was performed, and P<0.05 was considered statistically significant. Results: Animal experiments: Tanshinone ⅡA decreased the hearing threshold of DM group [(35.0±3.5) dB SPL vs. (55.3±8.1) dB SPL] (t=4.899, P<0.01), decreased the oxidative stress level in cochlea (t=4.384, P<0.05), improved the structure disorder, atrophy of cochlea vascular lines, vacuole increased phenomenon. Tanshinone ⅡA alleviated the increased permeability of the blood labyrinth barrier [Evans blue leakage (6.84±0.27) AU vs. (8.59±0.85) AU] in the cochlea of DM mice (t=2.770, P<0.05), reversed the apoptotic protein: Caspase3 (t=4.956, P<0.01) and Bax (t=4.388, P<0.05) in cochlear vascularis. Cell experiments: Tanshinone ⅡA decreased intracellular ROS content in a concentration-dependent way (t=3.569, P<0.05; t=4.772, P<0.01; t=7.494, P<0.01); Tanshinone ⅡA decreased apoptosis rate and apoptotic protein, and increased the expression of anti-apoptotic protein, p-PI3K/PI3K and p-AKT/AKT in concentration-dependent manner (all P values<0.05); LY294002 reversed the protective effect of tanshinone ⅡA on pericytes apoptosis (all P values<0.05). Conclusion: Tanshinone ⅡA can inhibit the apoptosis of cochlear pericytes induced by high glucose by reducing oxidative stress level and activating PI3K/AKT signaling pathway under high glucose environment, thus playing a protective role in diabetic hearing loss.

Clinical effects of free transplantation of expanded thoracodorsal artery perforator flaps in reconstructing cervical cicatrix contracture deformity after burns / 中华烧伤杂志

Objective: To explore the clinical effects of free transplantation of expanded thoracodorsal artery perforator flaps in reconstructing cervical cicatrix contracture deformity after burns. Methods: A retrospective observational study was conducted. From May 2018 to April 2021, 11 patients with cervical cicatrix contracture deformity after burns who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University, including 3 males and 8 females, aged 5 to 46 years, with a course of cervical cicatrix contracture deformity of 5 months to 8 years. The degree of cervical cicatrix contracture deformity was degree Ⅰ in one patient, degree Ⅱ in nine patients, and degree Ⅲ in one patient. In the first stage, according to the sizes of neck scars, one rectangular skin and soft tissue expander (hereinafter referred to as expander) with rated capacity of 200 to 600 mL was placed in the back. The expansion time was 4 to 12 months with the total normal saline injection volume being 3.0 to 3.5 times of the rated capacity of expander. In the second stage, free expanded thoracodorsal artery perforator flaps with areas of 10 cm×7 cm to 24 cm×13 cm were cut out to repair the wounds with areas of 9 cm×6 cm to 23 cm×12 cm which was formed after cervical cicatectomy. The main trunk of thoracodorsal artery and vein were selected for end-to-end anastomosis with facial artery and vein, and the donor sites were directly closed. The survival of flaps and healing of flap donor sites were observed on the 14th day post surgery. The appearances and cicatrix contracture deformity of the flaps, recovery of cervical function, and scar hyperplasia of donor sites were followed up. Results: On the 14th day post surgery, the flaps of ten patients survived, while ecchymosis and epidermal necrosis occurred in the center of flap of one patient and healed 2 weeks after dressing change. On the 14th day post surgery, the flap donor sites of 11 patients all healed well. During the follow-up of 6-12 months post surgery, the flaps of ten patients were similar to the skin around the recipient site in texture and color, while the flap of one patient was slightly swollen. All of the 11 patients had good recovery of cervical function and no obvious scar hyperplasia nor contracture in the flaps or at the donor sites. Conclusions: Application of expanded thoracodorsal artery perforator flaps can restore the appearance and function of the neck, and cause little damage to the donor site in reconstructing the cervical cicatrix contracture deformity after burns, which is worthy of clinical reference and application.

Effects of exosomes from human adipose-derived mesenchymal stem cells on pulmonary vascular endothelial cells injury in septic mice and its mechanism / 中华烧伤杂志

Objective: To investigate the effects of exosomes from human adipose-derived mesenchymal stem cells (ADSCs) on pulmonary vascular endothelial cells (PMVECs) injury in septic mice and its mechanism. Methods: The experimental research method was adopted. The primary ADSCs were isolated and cultured from the discarded fresh adipose tissue of 3 patients (female, 10-25 years old), who were admitted to the First Affiliated Hospital of Air Force Medical University undergoing abdominal surgery, and the cell morphology was observed by inverted phase contrast microscope on the 5th day. The expressions of CD29, CD34, CD44, CD45, CD73, and CD90 of ADSCs in the third passage were detected by flow cytometry. The third to the fifth passage of ADSCs were collected, and their exosomes from the cell supernatant were obtained by differential ultracentrifugation, and the shape, particle size, and the protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and β-actin of exosomes were detected, respectively, by transmission electron microscopy, nano-particle tracking analysis and Western blotting. Twenty-four adult male BALB/c mice were adopted and were divided into normal control group, caecal ligation perforation (CLP) alone group, and CLP+ADSC-exosome group with each group of 8 according to random number table (the same grouping method below) and were treated accordingly. At 24 h after operation, tumor necrosis factor (TNF-α) and interleukin 1β (IL-1β) levels of mice serum were detected by enzyme-linked immunosorbent assay, and lung tissue morphology of mice was detected by hematoxylin-eosin and myeloperoxidase staining, and the expression of 8-hydroxy-deoxyguanosine (8-OHdG) of mouse lung cells was detected by immunofluorescence method. Primary PMVECs were obtained from 1-month-old C57 mice regardless gender by tissue block method. The expression of CD31 of PMVECs was detected by immunofluorescence and flow cytometry. The third passage of PMVECs was co-cultured with ADSCs derived exosomes for 12 h, and the phagocytosis of exosomes by PMVECs was detected by PKH26 kit. The third passage of PMVECs were adopted and were divided into blank control group, macrophage supernatant alone group, and macrophage supernatant+ADSC-exosome group, with 3 wells in each group, which were treated accordingly. After 24 h, the content of reactive oxygen species in cells was detected by flow cytometry, the expression of 8-OHdG in cells was detected by immunofluorescence, and Transwell assay was used to determine the permeability of cell monolayer. The number of samples in above were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference t test. Results: The primary ADSCs were isolated and cultured to day 5, growing densely in a spindle shape with a typical swirl-like. The percentages of CD29, CD44, CD73 and CD90 positive cells of ADSCs in the third passage were all >90%, and the percentages of CD34 and CD45 positive cells were <5%. Exosomes derived from ADSCs of the third to fifth passages showed a typical double-cavity disc-like structure with an average particle size of 103 nm, and the protein expressions of CD9, CD63 and TSG101 of exosomes were positive, while the protein expression of β-actin of exosomes was negative. At 24 h after operation, compared with those in normal control group, both the levels of TNF-α and IL-1β of mice serum in CLP alone group were significantly increased (with t values of 28.76 and 29.69, respectively, P<0.01); compared with those in CLP alone group, both the content of TNF-α and IL-1β of mice serum in CLP+ADSC-exosome group was significantly decreased (with t values of 9.90 and 4.76, respectively, P<0.05 or P<0.01). At 24 h after surgery, the pulmonary tissue structure of mice in normal control group was clear and complete without inflammatory cell infiltration; compared with those in normal control group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP alone group were more obvious; compared with those in CLP alone group, the pulmonary tissue edema and inflammatory cell infiltration of mice in CLP+ADSC-exosome group were significantly reduced. At 24 h after operation, endothelial cells in lung tissues of mice in 3 groups showed positive expression of CD31; compared with that in normal control group, the fluorescence intensity of 8-OHdG positive cells of the lung tissues of mice in CLP alone group was significantly increased, and compared with that in CLP alone group, the fluorescence intensity of 8-OHdG positive cells in the lung tissues of mice in CLP+ADSC-exosome group was significantly decreased. The PMVECs in the 3rd passage showed CD31 positive expression by immunofluorescence, and the result of flow cytometry showed that CD31 positive cells accounted for 99.5%. At 12 h after co-culture, ADSC-derived exosomes were successfully phagocytose by PMVECs and entered its cytoplasm. At 12 h after culture of the third passage of PMVECs, compared with that in blank control group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant alone group was significantly increased (t=15.73, P<0.01); compared with that in macrophage supernatant alone group, the fluorescence intensity of reactive oxygen species of PMVECs in macrophage supernatant+ADSC-exosome group was significantly decreased (t=4.72, P<0.01). At 12 h after culture of the third passage of PMVECs, and the 8-OHdG positive fluorescence intensity of PMVECs in macrophage supernatant alone group was significantly increased; and compared with that in blank control group, the 8-OHdG positive fluorescence intensity of PMVECs in macrophage+ADSC-exosome supernatant group was between blank control group and macrophage supernatant alone group. At 12 h after culture of the third passage PMVECs, compared with that in blank control group, the permeability of PMVECs monolayer in macrophage supernatant alone group was significantly increased (t=6.34, P<0.01); compared with that in macrophage supernatant alone group, the permeability of PMVECs monolayer cells in macrophage supernatant+ADSC-exosome group was significantly decreased (t=2.93, P<0.05). Conclusions: Exosomes derived from ADSCs can ameliorate oxidative damage in mouse lung tissue, decrease the level of reactive oxygen species, 8-OHdG expression, and permeability of PMVECs induced by macrophage supernatant.

Clinical effect of large medium thickness skin graft on the back combined with autologous razor thick skin replantation to the back donor area in the treatment of giant congenital melanocytic nevus of the upper limb in children / 中国医师杂志

Objective:To investigate the clinical effect of large medium thickness skin graft on the back and scalp replantation in the back donor area after complete resection of giant congenital melanoma nevus (GCMN) in children′s upper limbs.Methods:From April 2017 to may 2020, 16 pediatric patients with GCMN of upper limbs were treated in the First Affiliated Hospital of Air Force Military Medical University, including 9 males and 7 females, aged from 2 years to 7 months to 12 years. Giant melanoma nevus area 14 cm×11 cm-23 cm×20 cm, the wound after removing the skin of giant melanocytic nevus of the limb was covered with vaseline oil gauze for 2-3 days, and then a large medium thickness skin graft was cut on the back with a drum skin extractor for transplantation. The wound in the back skin donor area was replanted with a blade thick scalp.Results:The effect of excision of giant nevus of upper limb and skin grafting on the wound of back medium thickness donor area in 16 pediatric patients was satisfactory, and there were no serious complications such as skin necrosis and poor survival. Plasma swelling was formed under the skin graft of one child′s limb, which healed after opening and drainage and three dressing changes. Anti-scar and rehabilitation treatment was performed on the limb and donor site.The patients were followed up for 6-18 months. There was no obvious scar hyperplasia and contracture in the skin graft area and donor area. The skin color and elasticity of the back and limb skin graft area were close to the normal skin around the wound, and the activities of elbow joint, wrist joint and interphalangeal joint were not limited. The parents of the pediatric were satisfied with the function and appearance of the limb skin graft area and back skin donor area of giant nevus.Conclusions:The function and appearance of large medium thickness skin graft on the back after excision of congenital giant nevus of upper limb in pediatric are better; There is no obvious scar formation after scalp replantation in the back donor area, and the repair effect is better.

Effect of microvascular pericytes of cochlear stria vascularis on endothelial cell permeability in C57BL/6J mice / 中华耳鼻咽喉头颈外科杂志

Objective: To study the changes in the permeability of the blood labyrinth barrier of the aging cochlea in mice, and to establish a non-contact co-culture model of endothelial cells (EC) and pericytes (PC) to furtherly investigate the cochlear stria vascularis microvascular pericytes impact on the permeability of endothelial cells. Methods: C57BL/6J mice were divided into two groups, three months old as young group, 12 months old as senile group. Cell experiment was divided into four groups, EC group, EC+PC co-culture group, D-gal+EC group and D-gal+EC+PC co-culture group. Auditory brainstem response (auditory brain response, ABR) was used to detect the auditory function of the two groups of mice. Evans blue staining was applied to detect the permeability of the cochlear blood labyrinth barrier of the two groups of mice. Transmission electron microscopy was used to observe the ultrastructure of blood labyrinth barrier endothelial cells, pericytes and tight junctions in the two groups of mice. Immunohistochemistry was used to detect the expression levels of tight junction proteins in the stria vascularis of the cochlea of the two groups of mice. Transwell chamber was used to detect the permeability of endothelial cells. Western blot and immunofluorescence technology were used to detect the expression level of tight junction protein on endothelial cells. SPSS 20.0 software was used to analyze the data. Results: Compared with the young group, the ABR threshold of the aging group was significantly increased, the latency of wave I was prolonged (t=10.25, P<0.01;t=5.61, P<0.05), the permeability of the cochlear blood labyrinth barrier was increased and the expression of tight junction protein on the vascular stria was decreased (P<0.05). The cochlear ultrastructure showed that the cochlear vascular stria microvascular lumen was deformed, the basement membrane thickened and the tight junction gap between endothelium enlarged. The positive rate of ECs and PCs in primary culture was more than 95%. The cells induced by 15 g/L D-gal were determined to be senescent cells. Compared with EC group, the expression of tight junction protein in endothelial cells of D-gal+EC group decreased（t=7.42，P<0.01；t=13.19，P<0.05）and the permeability increased (t=11.17, P<0.01). In the co-culture group, the expression of tight junction protein between endothelial cells in EC+PC co-culture group and D-gal+EC+PC co-culture group increased and the permeability decreased. Conclusions: In aging mice, the permeability of cochlear blood labyrinth barrier will increase and the level of tight junction protein will decrease; in aging state, cochlear vascular stria microvascular pericytes may affect endothelial cell permeability by regulating the expression of tight junction protein.

Short-term outcomes of total hip arthroplasty in the treatment of Tönnis grade 3 hip osteoarthritis in patients with spondyloepiphyseal dysplasia / 北京大学学报(医学版)

OBJECTIVE@#Severe hip osteoarthritis, caused by bone or joint maldevelopment, biomechanical transformation and previous surgical intervention, is inclusively existed in spondyloepiphyseal dysplasia (SED). To investigate and discuss the short-term efficacy and possible effects of total hip arthroplasty in the treatment of Tönnis grade 3 hip osteoarthritis in patients with SED.@*METHODS@#From January 2017 to June 2019, 374 patients with hip osteoarthritis were involved for total hip arthroplasty conducted by senior professional surgeons, of whom 9 patients (6 males and 3 females) with 12 hip osteoarthritis secondary to the SED met the inclusive and exclusive criteria and received the above-mentioned hip operation. The short-term outcomes were observed.@*RESULTS@#All the patients were implanted with Johnson & Johnson ceramic on ceramic cementless hip prostheses within the arthroplasty. They were followed up for an average period of 20 months. Except for one muscular calf vein thrombosis case, no complications, such as aseptic loosening, joint dislocation, fracture, neurovascular injury, deep vein thrombosis and infection were observed in all the 9 patients. Before the surgery, the average Harris hip score was 35.55, while the average of the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was 56.56. The level of quality of life indicated by SF-12 score was 41.56 on average. The mean pre-operation visual analogue scale (VAS) was 7.44. At the last follow-up, the average Harris hip score increased to 89.56, whereas the average WOMAC declined to 41.11. Compared with the baseline point, the average SF-12 score went up to 56.33. Dramatic drop of the mean VAS value to 2.67 was also observed at the last follow-up. In addition, post-operative increase of several pelvic-related parameters including pelvic incidence, pelvic tilt and sacral slope could be observed in the SED patients. The average measured pelvic incidence, pelvic tilt and sacral slope were 68.95°±4.60°, 52.75°±1.06° and 17.45°±1.77° before operation, respectively; whilst the mean value of these specific parameters increased to 76.98°±5.12°, 60.51°±4.35° and 18.10°±2.02°, respectively. The even leg lengths of the lower extremities were obtained after total hip arthroplasty.@*CONCLUSION@#Total hip arthroplasty is satisfactory in the short-term pain relieve and function recovery for the management of Tönnis grade 3 hip osteoarthritis secondary to the SED.

Effects of Ramipril on the expression of connexin 43 in cerebral arteries of spontaneously hypertensive rats / 生理学报

The present study was designed to examine whether Ramipril (an inhibitor of angiotensin-converting enzyme) affected spontaneous hypertension-induced injury of cerebral artery by regulating connexin 43 (Cx43) expression. Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) were randomly divided into WKY, WKY + Ramipril, SHR, and SHR + Ramipril groups (n = 8). The arterial pressure was monitored by the tail-cuff method, and vascular function in basilar arteries was examined by pressure myography. Hematoxylin-eosin (HE) staining was used to show vascular remodeling. The expression and distribution of Cx43 was determined by using immunofluorescence and immunohistochemistry analysis. The protein and mRNA levels of Cx43 were examined by Western blot and real-time PCR analysis, respectively. The results showed that chronic Ramipril treatment significantly attenuated blood pressure elevation (P < 0.01, n = 8) and blood vessel wall thickness in SHR (P < 0.01, n = 8). The cerebral artery contraction rate in the SHR group was higher than that in the WKY group (P < 0.05, n = 8). The cerebral artery contraction rate in the SHR + Ramipril group was lower than that in the SHR group (P < 0.05, n = 8). Pretreatment with 2-APB (Cx43 non-specific blocker) or Gap26 (Cx43 specific blocker) significantly decreased the vasoconstriction rate, while pretreatment with AAP10 (Cx43 non-specific agonist) significantly increased the vasoconstriction in the SHR + Ramipril group (P < 0.05, n = 8). In addition, the expression of Cx43 mRNA and protein in cerebral arteries of SHR group was higher than that of WKY group (P < 0.05, n = 8). The mRNA and protein expression of Cx43 in cerebral arteries of SHR + Ramipril group was significantly lower than that of SHR group (P < 0.05, n = 8). These results suggest that Ramipril can down-regulate the expression of Cx43 mRNA and protein in cerebral arterial cells of SHR, lower blood pressure, promote vasodilation, and improve arterial damage and vascular dysfunction caused by hypertension.

G protein-coupled estrogen receptor alleviates cerebral ischemia-reperfusion injury through inhibiting endoplasmic reticulum stress / 生理学报

The aim of this study was to investigate whether G protein-coupled estrogen receptor (GPER) could alleviate hippocampal neuron injury under cerebral ischemia-reperfusion injury (CIRI) by acting on endoplasmic reticulum stress (ERS). The CIRI animal model was established by middle cerebral artery occlusion (MCAO). Female ovariectomized (OVX) Sprague-Dawley (SD) female rats were randomly divided into 4 groups: control, ischemia-reperfusion injury (MCAO), vehicle (MCAO+DMSO), and GPER-specific agonist G1 (MCAO+G1) groups. The neurobehavioral score was assessed by the Longa score method, the morphological changes of the neurons were observed by the Nissl staining, the cerebral infarction was detected by the TTC staining, and the neural apoptosis in the hippocampal CA1 region was detected by TUNEL staining. The distribution and expression of GRP78 (78 kDa glucose-regulated protein 78) in the hippocampal CA1 region were observed by immunofluorescent staining. The protein expression levels of GRP78, Caspase-12, CHOP and Caspase-3 were detected by Western blot, and the mRNA expression levels of GRP78, Caspase-12, and CHOP were detected by the real-time PCR. The results showed that the neurobehavioral score, cerebral infarct volume, cellular apoptosis index, as well as GRP78, Caspase-12 and CHOP protein and mRNA expression levels in the MCAO group were significantly higher than those of control group. And G1 reversed the above-mentioned changes in the MCAO+G1 group. These results suggest that the activation of GPER can decrease the apoptosis of hippocampal neurons and relieve CIRI, and its mechanism may involve the inhibition of ERS.

The role and mechanism of TNFα-induced protein 3(A20) in phenotypic transformation of human dermal fibroblasts to myofibroblasts / 中国医师杂志

Objective To investigate the role and mechanism of A20 protein in the phenotypic transformation of human dermal fibroblasts (Fb) to myofibroblasts (MFb).Methods Primary human normal skin Fb (NFb) and scar skin Fb (HFb) were isolated and cuhured by enzymatic digestion.The cells were used in logarithmic growth phase at 3-5 passages,and Fb was stimulated by transforming growth factor-β (TGF-β) to mimic the Fb to MFb transformation process.Cell transfection was conducted to knock out the expression of A20 in Fb,and quantitative real-time polymerase chain reaction (PCR) test,protein immunoblotting test were conducted to detect A20,type Ⅰ collagen (Col Ⅰ),type Ⅲ collagen (Col Ⅲ)and α-smooth muscle (α-SMA),and p-Smad 2 expression in Fb.Results Compared with normal skin Fb,A20 was down-regulated in pathological scar Fb (P ＜ 0.05).After TGF-β stimulated,A20 expression was significantly decreased (P ＜ 0.05),while the expression of fibrosis associated molecules such as Col Ⅰ,Col Ⅲ and α-SMA and p-Smad 2 was significantly increased (P ＜ 0.05) in Fb.Knockout of A20 in Fb further promoted TGF-β induced the expression of fibrosis associated molecules such as Col Ⅰ,Col Ⅲ and o-SMA and p-Smad 2 (P ＜ 0.05).Conclusions A20 protein is involved in the process of human dermal Fb to MFb phenotype transformation,and A20 negatively regulates the transformation effect mediated by TGF-β by inhibiting the activation of Smad signaling pathway.

Progress and confusion in the diagnosis and treatment of scar / 中国医师杂志

The prevention and treatment of scars has always been an important task in the field of wound repair.Humans have started treatment for scars long time ago and have created a series of treatments.At the same time,with the development of science and technology,the diagnosis and treatment of scars have made great progress.Through the basic research for new therapeutic targets and innovative of treatment methods,treatment for scar turns more diversified and integrated.As a result,clinicians will suffer many challenges while having more options for scars treatment.Combined with traditional treatment,developing new treatments,and the comprehensive therapy of multiple treatment methods is the mainstream trend of scar treatment.

Effects of calcium-sensitive receptors on 11β-hydroxysteroid dehydrogenase type 2 and cortisol in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

OBJECTIVE@#To investigate the effects of calcium-sensitive receptors (CaSR) on the expression of 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) and cortisol concentration in a neonatal mouse model of persistent pulmonary hypertension (PPH).@*METHODS@#Fifty-six newborn C57BL/6 mice were randomly divided into a control group (n=14), a PPH group (n=14), an agonist group (n=14), and an inhibitor group (n=14). The mice in the PPH, agonist, and inhibitor groups were exposed to a 12% oxygen concentration, and the agonist group and inhibitor group were given CaSR agonist (GdCl3, 16 mg/kg) and CaSR antagonist (NPS2390, 1 mg/kg) intraperitoneally once a day, respectively. The mice in control group were exposed to air, and then injected with an equal volume of normal saline as those in the PPH group every day. All mice were treated for 14 days. Morphological examination of heart and lung tissues was performed using hematoxylin-eosin staining. The expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein in lung tissues were measured by qRT-PCR and Western blot respectively. Brain natriuretic peptide (BNP) and cortisol levels in lung tissues were determined using ELISA.@*RESULTS@#Compared with the control group, the PPH group had significantly increased pulmonary artery wall thickness (WT%), ratio of right to left ventricular thickness (RV/LV), alveolar mean linear intercept, and BNP concentration and a significantly reduced radial alveolar count (P<0.05); compared with the PPH group, the agonist group showed significant increases in WT% and BNP concentration, while the inhibitor group showed significant reductions in the two indicators (P<0.05). Compared with the control group, the PPH group showed significant reductions in the expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein, but a significant increase in cortisol concentration (P<0.05); compared with the PPH group, the agonist group had significantly lower expression levels of 11β-HSD2 mRNA and 11β-HSD2 protein, but a significant higher cortisol concentration, while the inhibitor group showed opposite changes in these indicators (P<0.05).@*CONCLUSIONS@#CaSR may control the development and progression of PPH in newborn mice by regulating the expression of 11β-HSD2 and cortisol concentration.

Effect of calcium-sensitive receptors on endothelial nitric oxide synthase and nitric oxide in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

OBJECTIVE@#To study the effect of calcium-sensitive receptors (CaSR) on the expression of endothelial nitric oxide synthase (eNOS) and the concentration of nitric oxide (NO) in a neonatal mouse model of persistent pulmonary hypertension (PPH).@*METHODS@#Eighty neonatal C57BL/6 mice were randomly divided into control, PPH, agonist and antagonist groups. The control group was exposed to air, and the other three groups were exposed to 12% oxygen. The agonist and antagonist groups were intraperitoneally injected with a CaSR agonist (GdCl 16 mg/kg) and a CaSR antagonist (NPS2390, 1 mg/kg), respectively, while the PPH and control groups were intraperitoneally injected with normal saline instead. All mice were treated for 14 days. Alveolar development and pulmonary vessels were assessed by hematoxylin-eosin staining. The protein and mRNA expression of eNOS and its localization in lung tissues were determined by Western blot, qRT-PCR and immunohistochemistry. The levels of brain natriuretic peptide (BNP) and NO in lung homogenate were determined using ELISA.@*RESULTS@#Compared with the control group, the PPH and agonist groups showed significant increases in alveolar mean linear intercept, the percent wall thickness of pulmonary arterioles, right to left ventricular wall thickness ratio (RV/LV) and BNP concentration, but a significant reduction in radial alveolar count (P<0.05). The antagonist group had significant improvements in all the above indices except RV/LV compared with the PPH and agonist groups (P<0.05). Compared with those in the control group, the protein and mRNA expression of eNOS and NO concentration were significantly increased in the PPH group and increased more significantly in the agonist group, but were significantly reduced in the antagonist group (P<0.05).@*CONCLUSIONS@#CaSR plays an important role in the development of PPH in neonatal mice, possibly by increasing eNOS expression and NO concentration.

Effects of free superficial temporal fascia flaps and skin grafts in repairing deep wounds in posterior ankle region of extensively burned patients / 中华烧伤杂志

Objective@#To observe the effects of the method of combining free superficial temporal fascia flaps with skin grafts in repairing deep wounds in posterior ankle region of extensively burned patients.@*Methods@#From September 2013 to February 2017, 11 extensively burned patients with deep tissue defects in posterior ankle region were treated in our unit. Two patients had tissue defects in bilateral posterior ankle regions. The wound sizes ranged from 5.8 cm×4.6 cm to 11.7 cm×5.2 cm. Free superficial temporal fascia flaps with the same sizes as the wounds were designed and resected to repair wounds in posterior ankle regions after debridement. The proximal end of superficial temporal veins and posterior tibial veins were performed with end-to-end anastomosis, and superficial temporal arteries and posterior tibial arteries were performed with end-to-side anastomosis. Skin grafts were resected to cover the superficial temporal fascia flaps according to patients′ condition of donor sites. The donor sites were sutured directly.@*Results@#All fascial flaps in 11 patients survived, while 2 skin grafts had partial necrosis, and they healed after secondary skin graft. All patients were followed up for 6 to 13 months, and the shape and function of the operation sites in all patients recovered well.@*Conclusions@#The method of combining free superficial temporal fascia flaps with skin grafts can repair deep wounds in posterior ankle regions of extensively burned patients. It has the advantages of good appearances in the recipient sites, strong resistances to infection of fascia flaps, minimal damages to the donor sites, short course of disease, and good prognosis of patients.

MiR-214 Regulates the Human Hair Follicle Stem Cell Proliferation and Differentiation by Targeting EZH2 and Wnt/β-Catenin Signaling Way In Vitro

miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, β-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/β-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/β-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR-214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, β-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, β-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of β-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR-214 directly targets EZH2 and affects Wnt/β-catenin signaling. The miR-214/EZH2/β-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.

Clinical study of early application of toe flap to repair thumb damaged electrical burns / 中国医师杂志

Objective To study the surgery effect on early application of toe flap to repair thumb severe electric bum.Methods From July 2007 to October 2017,a total of 25 cases of thumb severe electric burn were repaired by thumb toe flap grafting.The function of the finger after operation was evaluated comprehensively,and the effect of 72 hours before and after the injury was compared.Results All the flaps survived and the finger function was excellent.The excellent and good rate of 25 cases was 72％.The excellent and good rate of 72 hours before and after injury were 85.7％ and 54.5％ respectively.5 cases had bad wound complications after operation.1 cases were operated within 72 hours after injury,and 4 cases were operated after 72 hours.Conclusions The early application of toe flap to repair severe electric bum thumb can maximize the recovery of function of the thumb.The earlier the operation,the fewer complications,the better the repair effect.

Effects of long-term walking exercise on health status of low labor-intensity people / 中国医师杂志

Objective To investigate the effect of long-term walking exercise on the health status of low-labor-intensity population under long-term walking exercise.Methods A total of 100 volunteer teachers from universities in Beijing were enrolled.After physical examination,they were randomly divided into control group and observation group.The observation group regularly carried out corresponding encouragingactivities and recorded the daily number of steps.The corresponding indexes were measured again after 450 days.Results 45.24％ participants in the observation group got more than 75％ participation rate.The lower the participation rate,the lower the average daily steps (P ＜ 0.05).The fasting blood glucose,systolic blood pressure,LDL and total cholesterol were lower than other groups in the group participation rate ＜25％ (P ＜0,05).The body mass index,systolic blood pressure,fasting blood glucose,total cholesterol,ratio diabetes and hypertension in low activities group (＜ 7 500 steps/day) were lower than those in other groups (P ＜ 0.05).Conclusions Walking exercise can achieve good long-term persistence with constant feedback.For those who are older and whose initial indicators are higher,participation rate and exercise volume are more likely to be better under the guidance of encouragement.Persisting long-term walking exercise can help reduce blood pressure and blood lipids.

Strategy for wound repair of skin and soft tissue defect and systematic rehabilitation treatment for functional reconstruction of patients with severe burn or trauma on knees / 中华烧伤杂志

Objective@#Strategy for wound repair of skin and soft tissue defect and systematic rehabilitation treatment for functional reconstruction of patients with severe burn or trauma on knees.@*Methods@#From January 2015 to October 2016, 26 patients with skin and soft tissue defect on knees after severe burn or trauma were hospitalized in our unit. Among these patients, 14 patients had patellar ligament defect, and 16 patients had knee joint capsule defect. Wound debridement was operated on 1 to 3 days after admission. After debridement, the area of skin and soft tissue defect ranged from 10 cm×7 cm to 42 cm×18 cm. Vacuum sealing drainage (VSD) treatment was performed after debridement, and flap transplantation operation was performed after VSD treatment for 5 to 7 days. Defects of nine patients were treated with local rotation flaps. Seven patients with skin and soft tissue defects on knees and knee joint capsule defects of 5 cm×3 cm to 9 cm×7 cm were treated with free anterolateral femoral flaps combined with fascia lata. Ten patients with skin and soft tissue defects on knees and patellar ligament defects of 6 cm×3 cm to 12 cm×4 cm were treated with free anterolateral femoral flaps combined with iliotibial tract. The area of flaps ranged from 11 cm×9 cm to 22 cm×15 cm. After flap transplantation operation, functional reconstruction of knee joint was carried out according to early, continuous, and sequential systematic rehabilitation treatment strategy. The pain degree and function of knee joint of patients were scored by the International Knee Documentation Committee (IKDC) Knee Evaluation Form before operation and 12 months after operation. The knee joint flexion and extension degrees of patients were measured by joint protractor in 2 weeks and 12 months after operation. The color Doppler ultrasound was used to evaluate integrity of knee joint capsule and continuity of patellar ligament of patients in 6 and 12 months after operation.@*Results@#All flaps of 26 patients survived well, and wounds healed completely after the operation. Distal parts of flaps of 2 patients treated with free anterolateral femoral flaps had local necrosis after the operation, and their wounds healed after debridement and transplantation of autologous intermediate split-thickness skin graft of thigh. The IKDC Knee Evaluation Form score of patients was (79±8) points in 12 months after operation, which was significantly higher than (64±7) points before operation (t=7.20, P<0.05). The flexion degree of knee joint of patients was (117±10)° in 12 months after operation, which was significantly larger than (35±8)° in 2 weeks after operation (t=32.65, P<0.05). The extension degree of knee joint of patients was (12±9)° in 12 months after operation, which was significantly smaller than (61±9)° in 2 weeks after operation (t=19.63, P<0.05). In 6 and 12 months after operation, 9 patients treated with local rotation flaps had good integrity of knee joint capsule and continuity of patellar ligament; 7 patients treated with free anterolateral femoral flaps and fascia lata had good integrity of knee joint capsule; 10 patients treated with free anterolateral femoral flaps and iliotibial tract had good continuity of patellar ligament. During follow-up of 12 months, all flaps survived well; knees of all patients had good appearance; knee joints functioned normally.@*Conclusions@#Good appearance and function of knees can be achieved by repairing wound of skin and soft tissue defect on knees after severe burn or trauma with local rotation flaps or free anterolateral femoral flaps with fascia lata or iliotibial tract plus systematic rehabilitation treatment of knee joint in early stage after flap transplantation operation.

Prognostic analysis of plantar fasciitis treated by pneumatic ballistic extracorporeal shock wave versus ultrasound guided intervention / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the medium- and long-term effect of pneumatic ballistic extracorporeal shock wave versus ultrasound-guided hormone injection in the treatment of plantar fasciitis.</p><p><b>METHODS</b>The clinical data were collected from patients with plantar fasciitis admitted to PLA General Hospital pain department from September, 2015 to February, 2017. The patients were randomly divided into ultrasound-guided drug injection group and shock wave group. The therapeutic parameters including the numerical rating scale (NRS) scores in the first step pain in the morning, American Orthopedic Foot and Ankle Society (AOFAS) Ankle Hindfoot Scale, and thickness of the plantar fascia were monitored before and at 1 week, 1 month, 3 months, and 6 months after the treatment. The recurrence rate, effectiveness, and patient satisfaction were compared between the two groups at 6 months after the treatment.</p><p><b>RESULTS</b>Thirty-nine patients were enrolled in shock wave group and 38 patients in ultrasound group. The NRS scores in the first step pain in the morning were lowered after treatment in both groups (P<0.05), and the scores were significantly lower in ultrasound group than in shock wave group at 1 week and 1 month (P<0.01), but significantly higher in ultrasound group than in shock wave group at 3 and 6 months after treatment (P<0.05). The AOFAS functional scores were increased in both groups (P<0.05) at 6 months after treatment, was significantly lower in ultrasound group than in shock wave group than group B (90.44∓13.27 vs 75.76∓21.40; P<0.05). The effective rates in shock wave group and ultrasound group were 92.31% and 76.32%, respectively (P<0.05). Recurrence was found in 1 patient (2.56%) in shock wave group and in 8 (21.05%) in ultrasound group (P<0.05). The patient satisfaction scores were significantly higher in shock wave group than in ultrasound group (8.13∓2.67 vs 6.63∓3.75, P=0.048).</p><p><b>CONCLUSION</b>Pneumatic ballistic extracorporeal shock achieves better medium- and long-term outcomes than ultrasound-guided hormone injection in the treatment of plantar fasciitis.</p>

Role of calcium-sensing receptor in neonatal mice with persistent pulmonary hypertension / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.</p><p><b>METHODS</b>Forty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.</p><p><b>RESULTS</b>Compared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).</p><p><b>CONCLUSIONS</b>CaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.</p>

Influencing factors for complete resection and operation time in endoscopic submucosal dissection for colorectal tumors / 中华消化内镜杂志

Objective To explore influencing factors for complete resection and operation time of endoscopic submucosal dissection( ESD) for colorectal tumors. Methods This retrospective study included 95 consecutive colorectal tumors in 88 patients whose pathological diagnosis was adenoma and carcinoma, treated with ESD at the Department of Endoscopy of the First Hospital of Jilin University from January 2013 to December 2014. Multiple logistic regression analysis was conducted on the factors related to complete resection and operation time. Results Average tumor size was 28. 7±14. 1 mm(range,8?80 mm), and the average procedure time was 80. 72±63. 90 min. The rate of complete resection was 92. 6%(88/95),and the rate of incomplete resection was 7. 4%(7/95). Multivariate logistic regression analysis revealed that fibrosis (P=0. 012,OR=52. 473, 95%CI:2. 571?1140. 438) contributed to incomplete resection. Fibrosis ( P=0. 001, OR=0. 045, 95%CI:0. 007?0. 289) ,tumor size ( P=0. 035,OR=0. 170, 95%CI:0. 033?0. 884) ,granular?type laterally spreading tumor ( P=0. 013, OR=34. 432, 95%CI:2. 138?554. 476 ) , non?granular?type laterally spreading tumor(P=0. 044,OR=31. 715, 95%CI:1. 093?919. 904) were independent factors for extending operation time of colorectal ESD. Conclusion The severer fibrosis can induce higher rate of incomplete resection. The more severe fibrosis is, the larger tumor size is, and the longer operation time is.