[Inhibition of glutamatergic neurons in the dorsomedial periaqueductal gray alleviates excessive defensive behaviors of mice with post-traumatic stress disorder].

OBJECTIVE: To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey (dmPAG) in regulating excessive defensive behaviors in mice with post-traumatic stress disorder (PTSD). METHODS: Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno- associated viral vectors (rAAV2/9-CaMKII-mCherry, rAAV2/9-CaMKII-hM3Dq-mCherry and rAAV2/9-CaMKII-hM4Di-mCherry) into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons, followed 2 weeks later by PTSD modeling by single prolonged stress. The looming test, response to whisker stimulation test and contextual fear conditioning (CFC) test were used to observe changes in defensive behaviors of the PTSD mice. The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining. RESULTS: Compared with the control mice, the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest, response scores of defensive behaviors and freezing time (all P<0.01). Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors. Activation of the glutamatergic neurons in the dmPAG (in PTSD hM3Dq group) did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice, whereas inhibiting the glutamatergic neurons in the dmPAG (in PTSD hM4Di group) caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice (P<0.05 or 0.01). CONCLUSION: Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Comparison of different freezing methods for micro-volume semen.

BACKGROUND: Mico-volume semen freezing is essential and used popularly for fertility preservation of patients suffering cancer or undergoing male reproductive system related surgeries, and for other reasons that may risk fertility potential in ART cycles. However, clinicians and embryologists still face some unresolved technical and theoretical issues about the frozen-thawed efficiency. OBJECTIVE: To choose the appropriate freezing method for different volumes of normal semen samples. MATERIALS AND METHODS: We investigated the frozen-thawed outcomes of semen with different volumes (20 uL, 50 uL, 100 uL, 200 uL, 500 uL and 1 mL) using two freezing methods (FLNV, static liquid nitrogen vapour cooling followed by liquid nitrogen preservation; RFLN, direct rapid freezing in liquid nitrogen) and analyzed the vitality, progressive motility and DNA fragmentation index of thawed sperm. RESULTS: We found that semen freezing with volumes more than 100 uL had better outcomes than volumes less than or equal to 50 uL after thawing. FLNV presented a higher efficiency for cryopreservation of semen with volumes less than 50 uL. CONCLUSION: For smaller (micro) volumes, the FLNV technique is better than the RFLN method. https://doi.org/10.54680/fr24110110412.

[Influences and mechanism of extracellular vesicles from dermal papilla cells of mice on human hypertrophic scar fibroblasts].

Objective: To investigate the influences and mechanism of extracellular vesicles from dermal papilla cells (DPC-EVs) of mice on human hypertrophic scar fibroblasts (HSFs). Methods: The study was an experimental research. The primary dermal papilla cells (DPCs) of whiskers were extracted from 10 6-week-old male C57BL/6J mice and identified successfully. The DPC-EVs were extracted from the 3rd to 5th passage DPCs by ultracentrifugation, and the morphology was observed through transmission electron microscope and the particle diameter was detected by nanoparticle tracking analyzer (n=3) at 24 h after culture. The 3rd passage of HSFs were divided into DPC-EV group and phosphate buffer solution (PBS) group, which were cultured with DPC-EVs and PBS, respectively. The cell scratch test was performed and cell migration rate at 24 h after scratching was calculated (n=5). The cell proliferation levels at 0 (after 12 h of starvation treatment and before adding DPC-EVs or PBS), 24, 48, 72, and 96 h after culture were detected by using cell counting kit 8 (n=4). The protein expressions of α-smooth muscle actin (α-SMA) and collagen typeⅠ (ColⅠ) in cells at 24 h after culture were detected by immunofluorescence method and Western blotting, and the protein expression of Krüppel-like factor 4 (KLF4) in cells at 24 h after culture was detected by Western blotting. After the 3rd passage of HSFs were cultured with DPC-EVs for 24 h, the cells were divided into blank control group, KLF4 knockdown group, and KLF4 overexpression group according to the random number table. The cells in blank control group were only routinely cultured for 48 h. The cells in KLF4 knockdown group and KLF4 overexpression group were incubated with KLF4 knockdown virus for 24 h, then the cells in KLF4 knockdown group were routinely cultured for 24 h while the cells in KLF4 overexpression group were incubated with KLF4 overexpression virus for 24 h. The protein expressions of KLF4, α-SMA, and ColⅠ in cells were detected by Western blotting at 48 h after culture. Results: At 24 h after culture, the extracted DPC-EVs showed vesicular structure with an average particle diameter of 108.8 nm. At 24 h after scratching, the migration rate of HSFs in PBS group was (54±10)%, which was significantly higher than (29±8)% in DPC-EV group (t=4.37, P<0.05). At 48, 72, and 96 h after culture, the proliferation levels of HSFs in DPC-EV group were significantly lower than those in PBS group (with t values of 4.06, 5.76, and 6.41, respectively, P<0.05). At 24 h after culture, the protein expressions of α-SMA and ColⅠ of HSFs in DPC-EV group were significantly lower than those in PBS group, while the protein expression of KLF4 was significantly higher than that in PBS group. At 48 h after culture, compared with those in blank control group, the protein expression of KLF4 of HSFs in KLF4 knockdown group was down-regulated, while the protein expressions of α-SMA and ColⅠ were both up-regulated; compared with those in KLF4 knockdown group, the protein expression of KLF4 of HSFs in KLF4 overexpression group was up-regulated, while the protein expressions of ColⅠ and α-SMA were down-regulated. Conclusions: The DPC-EVs of mice can inhibit the proliferation and migration of human HSFs and significantly inhibit the expressions of fibrosis markers α-SMA and ColⅠ in human HSFs by activating KLF4.

[Perioperative management of wounds associated with secondary sternal osteomyelitis and/or mediastinitis after sternotomy and its clinical effects].

Objective: To investigate the perioperative management of wounds associated with secondary sternal osteomyelitis and/or mediastinitis after sternotomy, and to evaluate its clinical effects. Methods: This study was a retrospective observational study. From January 2017 to December 2022, 36 patients with wounds associated with secondary sternal osteomyelitis and/or mediastinitis after sternotomy who were conformed to the inclusion criteria were admitted to the Burn Center of PLA of the First Affiliated Hospital of Air Force Medical University, including 23 males and 13 females, aged 25 to 81 years. Preparation for surgery was made. For patients with suspected retrosternal mediastinal abscess cavity, all cancellous bone of the unhealed sternum was bitten off to fully expose the retrosternal mediastinum, remove the source of infection and granulation tissue, and to fill the sternum defect with flipped unilateral pectoralis major muscle. For patients who had no retrosternal mediastinal infection but had fresh granulation tissue in unhealed sternal wounds, the necrotic tissue and a small amount of necrotic sternum were palliatively removed, and bilateral pectoralis major muscles were advanced and abutted to cover the sternal defect. After the skin in the donor area was closed by tension-relieving suture, continuous vacuum sealing drainage was performed, and continuous even infusion and lavage were added 24 hours later. The thorax was fixed with an armor-like chest strap, the patients were guided to breathe abdominally, with both upper limbs fixed to the lateral chest wall using a surgical restraint strap. The bacterial culture results of wound exudation specimens on admission were recorded. The wound condition observed during operation, debridement method, muscle flap covering method, intraoperative bleeding volume, days of postoperative infusion and lavage, lavage solution volume and changes on each day, and postoperative complications and wound healing time were recorded. After discharge, the wound healing quality, thorax shape, and mobility functions of thorax and both upper limbs were evaluated during follow-up. The stability and closure of sternum were observed by computed tomography (CT) reexamination. Results: On admission, among 36 patients, 33 cases were positive and 3 cases were negative in bacterial culture results of wound exudation specimens. Intraoperative observation showed that 26 patients had no retrosternal mediastinal infection but had fresh granulation tissue in unhealed sternal wounds, palliative debridement was performed and bilateral pectoralis major muscles were advanced and abutted to cover the defect. In 10 patients with suspected retrosternal mediastinal abscess cavity, the local sternum was completely removed by bite and the defect was covered using flipped unilateral pectoralis major muscle. During the operation, one patient experienced an innominate vein rupture and bleeding of approximately 3 000 mL during mediastinal exploration, and the remaining patients experienced bleeding of 100-1 000 mL. Postoperative infusion and lavage were performed for 4-7 days, with a lavage solution volume of 3 500-4 500 mL/d. The lavage solution gradually changed from dark red to light red and finally clear. Except for 1 patient who had suture rupture caused by lifting the patient under the armpit during nursing on the 3rd day after surgery, the wounds of the other patients healed smoothly after surgery, and the wound healing time of all patients was 7-21 days. Follow-up for 3 to 9 months after discharge showed that the patient who had suture rupture caused by armpit lifting died due to multiple organ failure. In 1 patient, the armor-like chest strap was removed 2 weeks after surgery, and the shoulder joint movement was not restricted, resulting in local rupture of the suture, which healed after dressing change. The wounds of the remaining patients healed well, and they resumed their daily life. The local skin of patient's pectoralis major muscle defect was slightly sunken and lower than that of the contralateral thorax in the patients undergoing treatment of pectoralis major muscle inversion, while no obvious thoracic deformity was observed in patients undergoing treatment with pectoralis major muscle propulsion and abutment. The chest and upper limb movement in all patients were slightly limited or normal. CT reexamination results of 10 patients showed that the sternum was stable, the local sternum was closed or covered completely with no lacuna or defects. Conclusions: Once the wound associated with secondary sternal osteomyelitis and/or mediastinitis after sternotomy is formed, individualized and precise debridement should be performed as soon as possible, different transfer ways of pectoralis major muscle flap should be chosen to cover the defect, and postoperative continuous infusion and lavage together with strict thorax and shoulder joint restraint and immobilization should be performed. This treatment strategy can ensure good wound healing without affecting the shape and function of the donor area.

[Clinical effects of autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing middle urethral defect with penile defect].

Objective: To investigate the clinical effects of autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing middle urethral defect with penile defect. Methods: The retrospective observational study was conducted. Eight male patients (aged 14 to 58 years) with middle urethral defect and penile defect caused by various injuries who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University from January 2015 to January 2022. The length of urethral defect was 3 to 5 cm, and the wound area of penile defect after debridement was 5.0 cm×2.5 cm to 7.0 cm×5.5 cm. All the patients underwent autologous split-thickness skin grafting for prefabricating defect urethra in stage Ⅰ, and urethral anastomosis was performed and unilateral scrotal flap was transferred to reconstruct urethra and penis in stage Ⅱ. The area of scrotal flap was 6.0 cm×3.0 cm to 8.0 cm×6.0 cm. The wound in the donor area of skin graft was covered by oil gauze, and the wound of flap donor area was sutured directly. On the 7th day after the operation of stage Ⅱ, the survival of the flap was observed. In 3 weeks after the operation of stage Ⅱ, the urinary flow rate was measured by the urinary flow rate detector (urinary flow rate >15 mL/s was regarded as unobstructed urination), the urinary fistula and erectile function were observed, and the self-made therapeutic satisfaction questionnaire was used to investigate the therapeutic satisfaction degree of patients. During follow-up, the appearance of the flap recipient area was observed, the Vancouver scar scale (VSS) was used to evaluate the scar situation in the donor areas of skin graft and flap, the urinary flow rate was detected as before, the urethral stricture, urinary fistula, and erectile function were observed, and the therapeutic satisfaction degree of patients was investigated. Results: On the 7th day after the operation of stage Ⅱ, the flaps survived completely in 8 patients. In 3 weeks after the operation of stage Ⅱ, the urinary flow rate was 25.3 (18.0, 38.5) mL/s, with unobstructed urination, without urinary fistula and with erectile function, and the score of therapeutic satisfaction degree was 14.3 (14.0, 15.0). During follow-up of 1 to 7 years, the flap recipient area of 8 patients was full in appearance and not swollen, with similar color to the surrounding tissue; the VSS scores of the donor areas of skin graft and flap were 11.5 (10.0, 13.0) and 10.5 (9.3, 12.0), respectively, the urinary flow rate was 24.6 (17.7, 34.1) mL/s, with no urethral stricture, urinary fistula, and erectile dysfunction, and the score of therapeutic satisfaction degree was 13.5 (13.3, 14.8). Conclusions: Autologous split-thickness skin grafting for prefabricating urethra combined with scrotal flap in repairing the urethral and penile defects not only reconstructs the structure of urethra and the shape of penis, but also restores the sensation and erectile function of penis, with few postoperative complications, no obvious scar hyperplasia, and high satisfaction degree of patients, which is worthy of clinical promotion.

[Activation of GABAergic neurons in the zona incerta accelerates anesthesia induction with sevoflurane and propofol without affecting anesthesia maintenance or awakening in mice].

OBJECTIVE: To explore the regulatory effects of GABAergic neurons in the zona incerta (ZI) on sevoflurane and propofol anesthesia. METHODS: Forty-eight male C57BL/6J mice divided into 8 groups (n=6) were used in this study. In the study of sevoflurane anesthesia, chemogenetic experiment was performed in 2 groups of mice with injection of either adeno-associated virus carrying hM3Dq (hM3Dq group) or a virus carrying only mCherry (mCherry group). The optogenetic experiment was performed in another two groups of mice injected with an adeno-associated virus carrying ChR2 (ChR2 group) or GFP only (GFP group). The same experiments were also performed in mice for studying propofol anesthesia. Chemogenetics or optogenetics were used to induce the activation of GABAergic neurons in the ZI, and their regulatory effects on anesthesia induction and arousal with sevoflurane and propofol were observed; EEG monitoring was used to observe the changes in sevoflurane anesthesia maintenance after activation of the GABAergic neurons. RESULTS: In sevoflurane anesthesia, the induction time of anesthesia was significantly shorter in hM3Dq group than in mCherry group (P < 0.05), and also shorter in ChR2 group than in GFP group (P < 0.01), but no significant difference was found in the awakening time between the two groups in either chemogenetic or optogenetic tests. Similar results were observed in chemogenetic and optogenetic experiments with propofol (P < 0.05 or 0.01). Photogenetic activation of the GABAergic neurons in the ZI did not cause significant changes in EEG spectrum during sevoflurane anesthesia maintenance. CONCLUSION: Activation of the GABAergic neurons in the ZI promotes anesthesia induction of sevoflurane and propofol but does not affect anesthesia maintenance or awakening.

Investigation on oral nursing of elderly patients and prognosis of patients.

OBJECTIVE: To understand the oral care status of elderly inpatients in various departments, analyze the existing problems, and provide a basis for further improving the oral care practices and promoting the oral health of elderly patients. PATIENTS AND METHODS: This study intends to investigate the oral care status of patients in a tertiary hospital in Chongqing. This study was divided into two phases, the first phase was designed as a cross-sectional study. Our aim was to explore the implementation status of oral care in each department. The second phase of this study was to explore the correlation between patient oral care and patient outcomes. RESULTS: We extracted a total of 9,164 cases of elderly discharged patients. Primary care patients were mainly distributed in various general wards, among which orthopedics was the most frequent, accounting for 30.19%. The oral care doctor order rate of the patients with premium care was 80.21%, and the rate of oral care orders of the primary care patients was only 2.10%. The study analysis found that among surgical and intensive care unit (ICU) patients, patients in high-frequency group and low-frequency group were significantly better than that of patients without oral care in terms of overall survival. CONCLUSIONS: The oral care is still insufficient, and the frequency of use is relatively infrequent. This study also found that oral care can improve patient outcomes and reduce the incidence of ventilator-associated pneumonia (VAP).

[A study of the anti-fibrotic effect of ghrelin on mice with nonalcoholic steatohepatitis].

Objective: To study the anti-fibrotic effect of ghrelin on high-fat diet-induced non-alcoholic steatohepatitis (NASH) in mice. Methods: 24 male C57BL/6 mice were randomly divided into a normal diet group, a normal diet + ghrelin group, a high-fat diet group, and the high-fat diet + ghrelin group. The HFD and HFD+ghrelin groups were fed high-fat diet for 16 weeks to induce non-alcoholic steatohepatitis. Among them, the NCD+ghrelin group and HFD+ghrelin group were continuously given ghrelin intervention (11nmol·kg(-1)·d(-1)) for 2 weeks after feeding for 14 weeks. 16 mice were euthanized on weekends. The plasma levels of alanine aminotransferase (ALT) and hyaluronic acid (HA) were measured in mice. The content of hydroxyproline (Hyp) was determined in liver tissue. RT-qPCR was used to detect the mRNA expression levels of transforming growth factor ß1 (TGF-ß1) and collagen types I, III, and IV in liver tissue. A Western blot was used to detect the expression level of the α-smooth muscle actin (α-SMA) protein in liver tissue. HE staining was used to observe the morphological changes in liver tissue. VG staining was used to observe the fibrotic condition in liver tissue. Results: Compared with the NCD group, plasma ALT (266.80±146.80)U/L, HA (219.00±39.47) ng/ml levels, Hyp content (0.35±0.05)µg/mg prot (P < 0.05), mRNA expression levels of transforming growth factor ß1 (TGF-ß1) and collagen types I, III, IV (P < 0.05), and the expression level of α-SMA protein in the HFD group were significantly increased (P < 0.05), with congestion in the hepatic central lobular veins, hepatocytes swelling, and deposition of a large amount of collagen fibers in liver tissue. Compared with the HFD group, plasma ALT (57.17±20.88)U/L, HA (75.68±8.40)µg/mg levels, Hyp content (0.19±0.07)µg/mg prot, mRNA expression levels of transforming growth factor ß1 (TGF-ß1) and collagen types I, III, IV (P < 0.05), and the expression level of α-SMA protein in the HFD+ghrelin mice group was significantly reduced (P < 0.05), with only mild sinusoidal congestion in the liver tissue but significant improvement and reduction in liver injury and collagen fiber deposition. Conclusion: Ghrelin has a significant improvement effect on liver fibrosis in NASH mice.

[A prospective randomized controlled study on the repair of skin and soft tissue defect in functional areas of children with full-thickness skin grafts from different sites of abdomen].

Objective: To compare and analyze the effect of repairing small skin and soft tissue defect wounds in functional areas of children with full-thickness skin grafts from different sites of abdomen. Methods: A prospective randomized controlled study was conducted. From January 2019 to June 2020, 60 female children with small skin and soft tissue defects in functional areas requiring full-thickness skin grafting, who met the inclusion criteria, were admitted to the First Affiliated Hospital of Air Force Medical University. According to the random number table, the children were divided into two groups, with 28 cases left in lateral abdomen group aged 5 (3, 8) years and 29 cases in lower abdomen group aged 5 (3, 7) years after the exclusion of several dropped-out children in follow-up. In lower abdomen group, 20 (12, 26) cm2 wounds of children were repaired with (24±10) cm2 full-thickness skin graft from transverse skin lines in the inferior abdomen area, while in lateral abdomen group, 23 (16, 32) cm2 wounds of children were repaired with (24±9) cm2 full-thickness skin graft from below the umbilical plane to above the groin in the lateral abdomen area. All the children were treated with continuous intradermal suture at the donor site incision and received continuous negative pressure treatment of -10.64 to -6.65 kPa in the donor and recipient areas after operation. The donor site was treated with a medical skin tension-reducing closure device since post-surgery day (PSD) 7. The use of medical skin tension-reducing closure device at the donor site, postoperative complications and suture removal time of the donor area were recorded, and the incidence of complications was calculated. On PSD 7, a self-designed efficacy satisfaction questionnaire was used to investigate the parents' satisfaction with the curative effect of their children. In post-surgery month (PSM) 1 and 6, Vancouver scar scale (VSS) was used to evaluate the scar at the donor site, and the VSS score difference between the two time points was calculated; the scar width at the donor site was measured with a ruler, and the scar width difference between the two time points was calculated. Data were statistically analyzed with independent sample t test or Cochran & Cox approximate t test, Mann-Whitney U test, and Fisher's exact probability test. Results: The proportion of children in lateral abdomen group who used the medical skin tension-reducing closure device in the donor area for equal to or more than 4 weeks after surgery was significantly higher than that in lower abdomen group (P<0.05). On PSD 7, there was one case of partial incision dehiscence in the donor area, one case of peripheral skin redness and swelling in the donor area, and one case of fat liquefaction in the donor area in lateral abdomen group, and one case of partial incision dehiscence in the donor area in lower abdomen group. The incidence of postoperative complications at the donor site of children in lower abdomen group was significantly lower than that in lateral abdomen group (P<0.05). Compared with that in lateral abdomen group, the suture removal time at the donor site of children after surgery in lower abdomen group was significantly shorter (t'=17.23, P<0.01). On PSD 7, the satisfaction score of parents with the curative effect of their children in lower abdomen group was significantly higher than that in lateral abdomen group (t'=20.14, P<0.01). In PSM 1 and 6, the VSS scores of scar at the donor site of children in lower abdomen group were 2.7±0.9 and 2.8±1.0, respectively, which were significantly lower than 7.1±2.2 and 9.1±2.7 in lateral abdomen group (with t values of 10.00 and 11.15, respectively, P<0.01). In PSM 6, the VSS score of scar at the donor site of children in lateral abdomen group was significantly higher than that in PSM 1 (t=3.10, P<0.01), while the VSS score of scar at the donor site of children in lower abdomen group was not significantly higher than that in PSM 1 (P>0.05). The VSS score difference of scar at the donor site of children in lateral abdomen group was significantly greater than that in lower abdomen group (Z=-8.12, P<0.01). In PSM 1 and 6, the scar widths at the donor site of children in lower abdomen group were 2.0 (1.0, 2.0) and 2.0 (2.0, 3.0) mm, respectively, which were significantly narrower than 6.0 (4.0, 10.0) and 8.5 (5.0, 12.0) mm in lateral abdomen group (with Z values of -13.41 and -14.70, respectively, P<0.01). In PSM 6, the scar width at the donor site of children in lateral abdomen group was significantly wider than that in PSM 1 (Z=-2.79, P<0.01), while the scar width at the donor site of children in lower abdomen group was not significantly wider than that in PSM 1 (P>0.05). The difference of scar width at the donor site of children in lateral abdomen group was significantly greater than that in lower abdomen group (Z=-14.93, P<0.01). Conclusions: The use of full-thickness skin grafts from the lower abdomen to repair small skin and soft tissue defect wounds in functional areas of children, especially girls, is effective, simple and easy to operate, and conforms to the principle of aesthetic repair. Compared with transplantation with full-thickness skin graft from the lateral abdomen, lower abdominal full-thickness skin grafting has a low incidence of donor site complications and no obvious scar hyperplasia, which is worthy of clinical promotion.

[Clinical effects of bridge-type continuous negative pressure suction in postoperative fixation of upper limb soft tissue defect wound repaired with pedicled abdominal flap].

Objective: To observe the clinical effects of bridge-type continuous negative pressure suction in postoperative fixation of upper limb soft tissue defect wound repaired with pedicled abdominal flap. Methods: The retrospective observational study was conducted. From April 2018 to October 2020, ninety-five patients who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University, including 55 males and 40 females, aged 5-78 years, with a defect wound area of 82 (9, 216) cm2. All patients underwent abdominal flap repair for soft tissue defects of hand and forearm. According to the different fixation methods adopted in the operation area, the patients were divided into negative pressure group (n=48) and plaster group (n=47). Wounds of the injury sites of patients in the 2 groups were repaired by flap transplantation after debridement. The negative pressure suction device was placed after dry gauze dressing to form a "bridge" to fix the affected upper limb and chest and abdomen in negative pressure group. Patients in plaster group were treated with conventional dry gauze matting and plaster fixation. On post surgery day (PSD) 1, 3, 5, 7, and 14, flap blood circulation and pain intensity of patients in the 2 groups were calculated by self-made blood flow scoring scale and Changhai Pain Ruler, respectively. On PSD 5, the common complications in operative area and surrounding skin were observed and their incidences were calculated. On PSD 7, satisfaction degree of patients was scored. During follow-up of one month after pedicle division, the appearance and functional recovery of the affected limb were observed. Data were statistically analyzed with analysis of variance for repeated measurement, independent samples t est, Cochran & Cox approximate t test, chi-square test, and Wilcoxon rank-sum test. Results: On PSD 1, 3, 5, 7, and 14, the flap blood circulation scores of patients in negative pressure group did not change significantly, while that of patients in plaster group showed a time-dependent decrease, and the flap blood circulation scores of patients in negative pressure group were significantly higher than those in plaster group (t=2.259, 2.552, 2.784, 2.821, 3.003, P<0.05 or P<0.01). There were no significant changes in the pain intensity scores of patients in negative pressure group, while those of patients in plaster group increased in a time-dependent manner, and the pain intensity scores of patients in negative pressure group were significantly lower than those in plaster group (t=-4.818, -4.944, -5.011, -5.976, -6.721, P<0.05). On PSD 5, the incidences of common complications in operative area and surrounding skin of patients in negative pressure group were significantly lower than those in plaster group (χ2=6.773, 5.269, P<0.05 or P<0.01). On PSD 7, the satisfaction score of patients in negative pressure group was 14.7±1.1, which was significantly higher than 7.4±1.8 in plaster group (t=23.934, P<0.01). During follow-up of one month after pedicle division, the appearance and function of the affected limb of patients in the 2 groups recovered well. Conclusions: After repairing the upper limb soft tissue defect wound with pedicled abdominal flap, the bridge-type continuous negative-pressure suction fixation can effectively immobilize the affected limbs, chest and abdomen, reduce the incidence of common complications in the operative area and surrounding skin, relieve the pain of immobilization of patients, improve the blood circulation of flap and patient's satisfaction. Thus, it is an effective, portable, comfortable, and easy-to-operate method.

Child and Adolescent Psychosocial Support Programs Following Natural Disasters-a Scoping Review of Emerging Evidence.

PURPOSE OF REVIEW: This review aimed to identify and describe evidence published in the past 3 years from trials of psychosocial support programs for children and adolescents affected by natural disasters. RECENT FINDINGS: Previous reviews have indicated these programs are beneficial overall. Positive impacts were documented in school-based programs conducted by trained teachers and paraprofessionals with stronger effects achieved by more qualified professionals. The review found supporting evidence for positive impacts of post-disaster psychosocial programs. However, the strength of evidence is limited due to heterogeneity in interventions and evaluations. The stepped care model was found to be useful in differentiating between programs and level of available evidence. Hobfoll's five essential elements of mass trauma intervention provide an additional means of guiding program content and assessments, particularly for universal programs. Identified gaps in evidence included groups likely to be at most risk: preschool children, ethnically diverse groups, those with disability, and social disadvantage. There were promising indications of program benefits for groups with repeated exposure to natural disasters.

[A multicenter retrospective study on the etiology of necrotizing pneumonia in children].

Objective: To investigate the etiology of necrotizing pneumonia (NP) in children and the clinical characteristics of NP caused by different pathogens in China. Methods: A retrospective, case-control study was performed in children with NP who were admitted to 13 hospitals in China from January 2008 to December 2019. The demographic and clinical information, laboratory data, etiological and radiological findings were analyzed. The data were divided into three groups based on the following years: 2008-2011, 2012-2015 and 2016-2019, and the distribution characteristics of the pathogens in different period were compared. Meanwhile, the pathogens of pediatric NP in the southern and northern China were compared. And the clinical characteristics of the Mycoplasma pneumoniae (MP) NP and the bacterial NP were also compared. T-test or Mann-Whitney nonparametric test was used for comparison of numerical variables, and χ2 test was used for categorical variables. Results: A total of 494 children with NP were enrolled, the median ages were 4.7 (0.1-15.3) years, including 272 boys and 222 girls. Among these patients, pathogens were identified in 347 cases and the pathogen was unclear in the remaining 147 cases. The main pathogens were MP (238 cases), Streptococcus pneumoniae (SP) (61 cases), Staphylococcus aureus (SA) (51 cases), Pseudomonas aeruginosa (13 cases), Haemophilus influenzae (10 cases), adenovirus (10 cases), and influenza virus A (7 cases), respectively. MP was the most common pathogen in all three periods and the proportion increased yearly. The proportion of MP in 2016-2019 was significantly higher than that in 2012-2015 (52.1% (197/378) vs. 36.8% (32/87), χ2=6.654, P=0.010), while there was no significant difference in the proportion of MP in 2012-2015 and that in 2008-2011 (36.8% (32/87) vs. 31.0% (9/29), χ²=0.314, P=0.575).Regarding the regional distribution, 342 cases were in the southern China and 152 in the northern China. Also, MP was the most common pathogen in both regions, but the proportion of MP was higher and the proportion of SP was lower in the north than those in the south (60.5% (92/152) vs. 42.7% (146/342), χ2=13.409, P<0.010; 7.9% (12/152) vs. 14.3% (49/342), χ2=4.023, P=0.045). Comparing the clinical characteristics of different pathogens, we found that fever and cough were the common symptoms in both single MP and single bacterial groups, but chest pain was more common (17.0% (34/200) vs. 6.1% (6/98), χ2=6.697, P=0.010) while shortness of breath and wheezing were less common in MP group (16.0% (32/200) vs. 60.2% (59/98), χ2=60.688, P<0.01; 4.5% (9/200) vs. 21.4% (21/98), χ2=20.819, P<0.01, respectively). The white blood cell count, C-reactive protein and procalcitonin in the bacterial group were significantly higher than those in the MP group (14.7 (1.0-67.1)×109/L vs. 10.5 (2.5-32.2)×109/L, 122.5 (0.5-277.3) mg/L vs. 51.4 (0.5-200.0) g/L, 2.13 (0.05-100.00) µg/L vs. 0.24 (0.01-18.85) µg/L, Z=-3.719, -5.901 and -7.765, all P<0.01). Conclusions: The prevalence of pediatric NP in China shows an increasing trend during the past years. MP, SP and SA are the main pathogens of NP, and the most common clinical symptoms are fever and cough. The WBC count, C-reactive protein and procalcitonin in bacterial NP are significantly higher than those caused by MP.

The Potential Impact and Usability of the Eighth Edition TNM Staging Classification in Oral Cavity Cancer.

AIMS: In the current eighth edition head and neck TNM staging, extranodal extension (ENE) is an adverse feature in oral cavity squamous cell cancer (OSCC). The previous seventh edition N1 with ENE is now staged as N2a. Seventh edition N2+ with ENE is staged as N3b in the eighth edition. We evaluated its potential impact on patients treated with surgery and postoperative intensity-modulated radiotherapy (IMRT). MATERIALS AND METHODS: OSCC patients treated with primary surgery and adjuvant (chemo)radiotherapy between January 2005 and December 2014 were reviewed. Cohorts with pathological node-negative (pN-), pathological node-positive without ENE (pN+_pENE-) and pathological node-positive with ENE (pN+_pENE+) diseases were compared for local control, regional control, distant control and overall survival. The pN+ cohorts were further stratified into seventh edition N-staging subgroups for outcomes comparison. RESULTS: In total, 478 patients were evaluated: 173 pN-; 159 pN+_pENE-; 146 pN+_pENE+. Outcomes at 5 years were: local control was identical (78%) in all cohorts (P = 0.892), whereas regional control was 91%, 80% and 68%, respectively (P < 0.001). Distant control was 97%, 87%, 68% (P < 0.001) and overall survival was 75%, 53% and 39% (P < 0.001), respectively. Overall survival for N1 and N2a subgroups was not significantly different. In the seventh edition N2b subgroup of pENE- (n = 79) and pENE+ (n = 79) cohorts, overall survival was 67% and 37%, respectively. In the seventh edition N2c subgroups, overall survival for pENE- (n = 17) and pENE+ (n = 38) cohorts was 65% and 35% (P = 0.08), respectively. Overall, an additional 128 patients (42% pN+) were upstaged as N3b. CONCLUSIONS: When eighth edition staging was applied, stage migration across the N2-3 categories resulted in expected larger separations of overall survival by stage. Patients treated with primary radiation without surgical staging should have outcomes carefully monitored. Strategies to predict ENE preoperatively and trials to improve the outcomes of pENE+ patients should be explored.

[Clinical application effect of latissimus dorsi muscle flap in reconstruction of muscle strength around shoulder after electric burns].

Objective: To investigate the clinical application effect of latissimus dorsi muscle flap in reconstruction of muscle strength around shoulder after electric burns. Methods: From March 2014 to September 2020, 13 patients with electric burns and severe injury around shoulder were admitted to the First Affiliated Hospital of Air Force Medical University, including 11 males and 2 females, aged 19-55 years. A retrospective observational study was conducted. The left upper limbs were injured in 8 cases, and the right upper limbs were injured in 5 cases, all with eschar wounds of Ⅲ-Ⅳ degree. Among which, there were biceps defects in 6 cases, deltoid defects in 3 cases, triceps defects in 2 cases, and composite defects of multiple muscles around shoulder in 2 cases. The surgery was carried out in two stages. In stage Ⅰ, debridement and exploration of electric burn wounds around shoulders were conducted to preserve local tissue and save the limb as much as possible on the premise of guaranteeing the stability of the body condition. After the last debridement, the wound area was from 10 cm×6 cm to 40 cm×15 cm, the muscle defect area was from 8 cm×4 cm to 19 cm×12 cm, and the humerus was exposed in 7 patients. In stage Ⅱ, according to the residual limb defect degree, muscle reconstruction around shoulder was conducted with the latissimus dorsi muscle flap, and area of the latissimus dorsi muscle flap was 15 cm×6 cm to 20 cm×18 cm. The residual wounds were repaired with autologous split-thickness skin grafts of head, and the donor sites of muscle flaps were sutured directly. The survivals of the muscle flaps and wounds closure post operation, and the appearances of the donor sites and recipient sites during follow-up were observed. At the last follow-up, the shoulder joint function was evaluated using the trial standard for the evaluation of the functions of the upper limbs of the Hand Surgery Society of the Chinese Medical Association, and the satisfaction degrees of patients for appearance and function recoveries of shoulder were investigated by self-made questionnaire with reference to the concise test scoring system of shoulder joint. Results: All of the 13 muscle flaps around shoulder survived after surgery. Two patients had residual wounds in the skin grafting area, the wound in one of the patients was healed after dressing change, and the wound in the other 1 patient was healed with the second autologous split-thickness skin grafting on head after dressing change. During follow-up of 6 to 18 months for all the patients, the muscle flaps of patients were full in appearance and not bloated, and atrophic scar in the repaired area was soft in texture and closed with normal skin around. Linear suture scars were left in the donor sites of muscle flaps, which did not affect the overall appearance. At the last follow-up, the active abduction range of the shoulder joint was 60-90°, upward lift on 120-180°, muscle strength recovered to level Ⅳ and above in 8 cases and to level Ⅲ in 5 cases, and the shoulder joint function was evaluated as excellent in 8 cases and good in 5 cases; 10 patients were very satisfied and 3 patients were satisfied with the appearance and function recovery of the shoulders. Conclusions: The application of latissimus dorsi muscle flap provides a better choice for the muscle strength reconstruction around shoulder after electric burns, with good appearance of the operative areas and ideal prognosis of upper limb function.

[Clinical effects of expanded flap in repairing the wounds with exposed titanium mesh after cranioplasty with titanium mesh].

Objective: To explore the clinical effects of expanded flap made by skin and soft tissue expander (hereinafter referred to as expander) in repairing the wounds with exposed titanium mesh after cranioplasty with titanium mesh. Methods: A retrospective observational study was conducted. From April 2015 to October 2019, 13 patients with wounds with exposed titanium mesh after cranioplasty with titanium mesh were admitted to the First Affiliated Hospital of Air Force Medical University, including 10 males and 3 females, aged 18 to 70 years. Exposure of titanium mesh occurred 3 months to 4 years after cranioplasty with titanium mesh. The wound area of exposed titanium mesh ranged from 1.5 cm×0.6 cm to 6.3 cm×6.0 cm. In the first stage, one or two square expanders with rated capacity of 50-200 mL were placed under the normal scalp 1 cm away from the edge of the wound surface of exposed titanium mesh. The water injection time was 2 to 3 months with the total water injection volume being 1.6 to 2.0 times of the rated capacity of expander. In the second stage, the expander was removed and the expanded flap (size ranging from 4.1 cm×1.8 cm to 9.1 cm×7.9 cm) was transferred to repair the wound of exposed titanium mesh. The placement site of the expander, the transfer form of the expanded flap, the postoperative wound healing of the titanium mesh exposed site, and the survival of expanded flap were recorded. The scar of the head incision and the appearance of head were followed up. Results: Among the patients in this group, the expanders of 7 were placed in the temporal region, while the other 6 were placed at the top. The 11 patients were treated with advanced expanded flap, while the other 2 patients were treated with pedicled expanded flap. The head wounds of patients in this group successfully healed with retaining of the titanium mesh. The wound was healed after dressing change in 1 patient with necrosis at the tip of the expanded flap. The expanded flaps of 12 patients survived after transfer. Patients in this group were followed up for 12 months after surgery, the exposed titanium meshes were retained, the incisions healed well with the scars concealed, the hair on the scalp grew well, and the appearance of head was comparatively good. Conclusions: Using expanded flap in the repair of the wounds with exposed titanium mesh after cranioplasty with titanium mesh can effectively cover the wound and retain the titanium mesh, achieving good function and appearance.

[A new lossy compression method for fetal heart rate signals-Convolutional Codec Network].

In order to reduce the energy loss during data transmission and storage in the Internet of Things system and improve the transmission efficiency of fetal heart rate data to allow real-time monitoring of the fetus, we used a convolutional codec network (CC-Net) to compress the data. The network has two modules: the encoding and decoding modules. The original data are compressed in the encoding module and reconstructed in the decoding module. The internal parameters are continuously updated using the mean square error of the original and the reconstructed signals to minimize the error to obtain effectively compressed data in the encoding module. In this study, the compression ratio of fetal heart rate signals using this method reached 12.07%, and the error between the reconstructed and original signals was 0.03. The proposed CC-Net can achieve a very low compression ratio for fetal heart rate compression while ensuring a high similarity between the reconstructed and the original signals to retain important information in fetal heart rate signals.

Denoising method for Raman spectra with low signal-to-noise ratio based on feature extraction.

Raman spectroscopy is a non-destructive technique utilizing lasers to observe scattered light in order to determine things such as vibrational modes in the molecular system. A major problem inherent to this technique is that due to their short exposure time and the low power of the excitation laser, Raman signals are very weak. They tend to be much weaker than the noise and can even be drowned out. Conventional denoising methods are currently unable to extract Raman peaks with precision so it is necessary to specifically study Raman signal extraction methods that involve a low signal-to-noise ratio (SNR). In this study, a denoising method for Raman spectra with low SNR based on feature extraction was proposed. Based on the Hilbert Vibration Decomposition (HVD) method, the Raman spectra was decomposed into two components. The peaks were located in the first component and compensated by those in the second component. Then based on the position and height of the peaks, their full widths at half maximum (FWHM) are calculated. Finally, based on the position, height and FWHM of the peaks, Gaussian signals are used to reconstruct the Raman peaks from strong noise and baseline. In the data simulation experiment, the denoising method used improved the SNR from 3.5316 to 130.6386 and the mean square error (MSE) was reduced from 213.8635 to 14.0404. In the actual experiment, this method successfully extracted the characteristic peaks of melamine despite the noise from employing a low excitation laser (10 mW). The characteristics such as the amplitude and position of the peaks were identical to those obtained under a high excitation laser (150 mW). The error of the FWHM under different excitation laser powers (10 and 150 mW) was less than the spectral resolution. Using the method proposed in this paper, the Raman signal of biological samples such as rice leaves were extracted from the raw spectrum, and information on the spectral peak position, amplitude and FWHM were obtained with clarity. The characteristic peaks of the carotene molecule, protein amide I, protein phenylalanine, nucleic acid cytosine, cellulose, DNA phosphodiester, RNA phosphodiester, D-glucose, α-D glucose, chlorophyll, lignin and cellulose were all accurate as well. The results from the simulation data and actual experiments show that a method based on feature extraction can effectively extract Raman peaks even when they are submerged in background noise. It should be noted that the practicality of this method lies in the fact that it requires few parameters and is simple to operate and implement.

Correlations of the expression of γδ T cells and their co-stimulatory molecules TIGIT, PD-1, ICOS and BTLA with PR and PIBF in the peripheral blood and decidual tissues of women with unexplained recurrent spontaneous abortion.

Semi-allogeneic embryos are not rejected by the maternal immune system due to maternal-fetal immune tolerance. Progesterone (P) receptor (PR)-expressing γδ T cells are present in healthy pregnant women. In the presence of P, these cells secrete an immunomodulatory protein called progesterone-induced blocking factor (PIBF), which can facilitate immune escape and is important in preventing embryonic rejection. This work investigated the correlations of the expression of γδ T cells and their co-stimulatory molecules T cell immunoglobulin and ITIM domain (TIGIT), programmed cell death 1 (PD-1), inducible co-stimulator (ICOS) and B and T lymphocyte attenuator (BTLA) with progesterone receptor (PR) and progesterone-induced blocking factor (PIBF) in peripheral blood and decidual tissue in women with unexplained recurrent spontaneous abortion (URSA) and normal pregnant (NP) women. We confirmed that γδ T cell proportions and PIBF expression in the peripheral blood and decidua of URSA women decreased significantly, while PR expression in decidua decreased. However, TIGIT, PD-1, ICOS and BTLA expression in γδ T cells in peripheral blood did not change, while TIGIT and PD-1 expression in γδ T cells in decidua increased significantly. Under the action of PHA-P (10 µg/ml), co-blocking of TIGIT (15 µg/ml) and PD-1 (10 µg/ml) antibodies further induced γδ T cell proliferation, but PIBF levels in the culture medium supernatant did not change. At 10-10 M P, γδ T cells proliferated significantly, and PIBF concentrations in the culture medium supernatant increased. γδ T cells co-cultured with P, TIGIT and PD-1 blocking antibodies showed the most significant proliferation, and PIBF concentrations in the culture medium supernatant were the highest. These results confirm that P is necessary for PIBF production. The TIGIT and PD-1 pathways participate in γδ T cell proliferation and activation and PIBF expression and play important roles in maintaining pregnancy.