[Clinical and genetic analysis of a patient with HUPRA syndrome due to missense variants of SARS2 gene and literature review].

Objective: To explore the clinical manifestations and genotype of an infant with hyperuricemia, pulmonary hypertension, renal failure in infancy, and alkalosis syndrome (HUPRAS). Methods: Clinical data of the patient were collected. Peripheral blood samples from the patient and his parents were acquainted for whole exome sequencing. The filtrated variants were verified by Sanger sequencing. The pathogenicity of the variants was predicted by bioinformatic tools. Results: The patient is a male infant of 6 months old, carrying two missense variants in the SARS2 allele: a paternal inherited c.1205G>A (p. Arg402His) and a maternal inherited c.680G>A (p. Arg227Gln). The two variants were in extremely low population frequencies. The pathogenetic prediction tools categorized them as deleterious. Arg402 and Arg227 were highly conserved in evolution. The variants led to changes in the hydrogen bonds and hydrophobicity of seryl-tRNA synthetase encoded by SARS2. Conclusions: c.1205G>A (p. Arg402His) and c.680G>A (p. Arg227Gln) are the possible causative variants of the HUPRA syndrome.

[Association of sleep duration and physical exercise with dyslipidemia in older adults aged 80 years and over in China].

Objective: To explore the impact of sleep duration, physical exercise, and their interactions on the risk of dyslipidemia in older adults aged ≥80 (the oldest old) in China. Methods: The study subjects were the oldest old from four rounds of Healthy Aging and Biomarkers Cohort Study (2008-2009, 2011-2012, 2014 and 2017-2018). The information about their demographic characteristics, lifestyles, physical examination results and others were collected, and fasting venous blood samples were collected from them for blood lipid testing. Competing risk model was used to analyze the causal associations of sleep duration and physical exercise with the risk for dyslipidemia. Restricted cubic spline (RCS) function was used to explore the dose-response relationship between sleep duration and the risk for dyslipidemia. Additive and multiplicative interaction model were used to explore the interaction of sleep duration and physical exercise on the risk for dyslipidemia. Results: The average age of 1 809 subjects was (93.1±7.7) years, 65.1% of them were women. The average sleep duration of the subjects was (8.0±2.5) hours/day, 28.1% of them had sleep duration for less than 7 hours/day, and 27.2% had sleep for duration more than 9 hours/day at baseline survey. During the 9-year cumulative follow-up of 6 150.6 person years (follow-up of average 3.4 years for one person), there were 304 new cases of dyslipidemia, with an incidence density of 4 942.6/100 000 person years. The results of competitive risk model analysis showed that compared with those who slept for 7-9 hours/day, the risk for dyslipidemia in oldest old with sleep duration >9 hours/day increased by 22% (HR=1.22, 95%CI: 1.07-1.39). Compared with the oldest old having no physical exercise, the risk for dyslipidemia in the oldest old having physical exercise decreased by 33% (HR=0.67, 95%CI: 0.57-0.78). The RCS function showed a linear positive dose-response relationship between sleep duration and the risk for hyperlipidemia. The interaction analysis showed that physical exercise and sleep duration had an antagonistic effect on the risk for hyperlipidemia. Conclusion: Physical exercise could reduce the adverse effects of prolonged sleep on blood lipids in the oldest old.

[Humanized anti-CD25 monoclonal antibody as a salvage therapy for steroid-refractory acute graft-versus-host disease after hematopoietic stem cell transplantation].

Objective: To investigate the efficacy of humanized anti-CD25 monoclonal antibody for steroid-refractory acute graft-versus-host disease (SR-aGVHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) recipients. Methods: A total of 64 patients with SR-aGVHD between June 2019 and October 2020 in Suchow Hopes Hematology Hospital were enrolled in this study. Humanized anti-CD25 monoclonal antibodies 1 mg·kg(-1)·d(-1) were administered on days 1, 3, and 8, and then once per week according to the disease progression. Efficacy was assessed at days 7, 14, and 28 after humanized anti-CD 25 treatment. Results: Of the 64 patients with a median age of 31 (15-63) years, 38 (59.4%) were male and 26 (40.6%) were female. The overall response (OR) rate of the humanized CD25 monoclonal antibody in 64 patients with SR-aGVHD on days 7, 14, and 28 were 48.4% (31/64), 53.1% (34/64), and 79.7% (51/64), respectively. Liver involvement is an independent risk factor for poor efficacy of humanized CD25 monoclonal antibody for SR-aGVHD at day 28 (OR=9.588, 95% CI 0.004-0.291, P=0.002). The median follow-up time for all patients was 17.1 (0.2-50.8) months from the start of humanized CD25 monoclonal antibody therapy. The 1- and 2-year OS rates were 63.2% (95% CI 57.1% -69.3%) and 52.6% (95% CI 46.1% -59.1%), respectively. The 1- and 2-year DFS rates were 58.4% (95% CI 52.1% -64.7%) and 49.8% (95% CI 43.4% -56.2%), respectively. The 1- and 2-year NRM rates were 28.8% (95% CI 23.1% -34.5%) and 32.9% (95% CI 26.8% -39.0%), respectively. The results of the multifactorial analysis showed that liver involvement (OR=0.308, 95% CI 0.108-0.876, P=0.027) and GVHD grade Ⅲ/Ⅳ (OR=9.438, 95% CI 1.211-73.577, P=0.032) were independent risk factors for OS. Conclusion: Humanized CD25 monoclonal antibody has good efficacy and safety for SR-aGVHD. This study shows that SR-aGVHD with pretreatment grade Ⅲ/Ⅳ GVHD and GVHD involving the liver has poor efficacy and prognosis and requires early intervention.

[Clinical effect of the giant deep inferior epigastric artery paraumbilical perforator flap in repairing the circular high-voltage electric burn wounds on the wrist].

Objective: To investigate the clinical effect of the giant deep inferior epigastric artery paraumbilical perforator flap in repairing the circular high-voltage electric burn wounds on the wrist. Methods: A retrospective observational study method was used. From September 2016 to October 2021, thirteen male patients (aged 20-43 years) with annular high voltage (10-100 kV) electrical burns on the wrist were admitted to the Beijing Jishuitan Hospital. At the early stage after injury, the patient's wrist was subjected to incision, tension reduction and debridement, with the wound area after debridement being 27 cm×16 cm-32 cm×19 cm; in 12 patients with vascular injury, the radial or ulnar artery was reconstructed by great saphenous vein transplantation, with the length of 15-25 cm; the wrist wound was repaired by free transplantation of the deep inferior epigastric artery paraumbilical perforator flap (if the wound was giant, the lower abdominal flap carrying other perforators was used), with the area of 30 cm×19 cm-35 cm×20 cm. The donor site was repaired by direct suture+skin grafting or relay flap transplantation. After surgery, the survival of flap in recipient area, as well as survival of the skin or flap in donor site were observed. During follow-up, the appearances of the flap in recipient area and the recovery of hand function, as well as the healing of donor site, occurrence of abdominal wall hernia, and scar in skin graft area were observed. Results: After surgery, all the 13 patients' paraumbilical perforator flaps survived. Among them, 3 patients had subcutaneous fat necrosis at the distal end of the wrist flap, and the wound had mild infection, which healed after re-expansion and dressing change. All the skin grafts in the donor site of 10 patients survived, and the flaps in the donor site of 3 patients survived well. The patients were followed up for 6 months to 3 years. The flaps in recipient area were in good shape, 8 cases had partial recovery of hand function, and 5 cases had loss of finger flexion function; the donor site of abdominal flap healed well with no abdominal hernia occurred, and the skin graft site had no obvious scar hyperplasia and was soft in texture. Conclusions: Early vascular reconstruction after injury, together with free transplantation of the giant deep inferior epigastric artery paraumbilical perforator flap are effective in repairing circular high-voltage electrical burn wounds on the wrist.

Intermittent fasting in type 2 diabetes: from fundamental science to clinical applications.

Type 2 diabetes mellitus (T2DM) is a huge challenge for global public health systems. Currently, healthcare policies advocate the prevention of the onset and progression of T2DM by improving individual lifestyles. The increasing benefits of intermittent fasting (IF) as a dietary intervention have been elucidated. However, the beneficial effects of IF in T2DM remain inconclusive. We demonstrated the physiological mechanisms underlying the positive effects of IF in T2DM. IF could trigger metabolic transformation to improve systemic metabolism and induce tissue-specific metabolic adaptations through alterations in the gut microbiota, adipose tissue remodeling, correction of circadian rhythm disturbances, and increased autophagy in peripheral tissues. The efficacy and safety of IF regimens in clinical applications carry a risk of hypoglycemia and require monitoring of blood glucose and timely adjustment of medications. However, there is limited evidence of a positive effect of IF in weight loss and improvement of glycemic variables. Overall, IF serves as a promising therapeutic target for T2DM and needs to be established by a large randomized controlled trial.

[Current status of influencing factors for postoperative anastomotic leakage in low rectal cancer].

The incidence of anastomotic leakage, a common and serious postoperative complication of low rectal cancer, remains high. Clarifying the risk factors for anastomotic leakage in patients with low rectal cancer after surgery can help guide clinical treatment and help patients improve their prognosis. The current literature suggests that the risk factors affecting the occurrence of anastomotic leakage after low rectal cancer include three aspects: (1) individual factors: male gender, high body mass index, malnutrition, smoking, alcoholism, and metabolic diseases; (2) tumor factors: the lower margin of tumor <5 cm from the anal verge, tumor diameter >2.5 cm, late tumor stage, high level of tumor markers and preoperative intestinal obstruction; (3) surgical factors: long operative time (>180 min), intraoperative bleeding (≥70 ml), more than 2 cartridges of stapling for anastomosis, contamination of the operative field, epidural analgesia and intraoperative hypothermia. Notably, the surgical approach (laparoscopic, open and hand-assisted laparoscopic surgery) was not a factor influencing the occurrence of postoperative anastomotic leakage in low rectal cancer. The findings on the effects of receiving neoadjuvant therapy, gut microbiota,intestinal bowel preparation, insufficient time for preoperative antibiotic prophylaxis, left colonic artery dissection, intraoperative blood transfusion, pelvic drainage, transanal drainage and combined organ resection, and postoperative diarrhea on postoperative anastomotic leakage in low rectal cancer are controversial. However, clinical workers can still take measures to reduce the risk of anastomotic leakage according to the above risk factors by making a good assessment before surgery, actively avoiding them during and after surgery, and taking measures for each step, so as to bring maximum benefits to patients.

[Analysis of cervical instability and clinical characteristics in treatment-naive rheumatoid arthritis patients].

OBJECTIVE: To study the proportion of cervical spine instability in treatment-naive rheumatoid arthritis (RA) patients, to investigate the associated neck symptoms, and to analyze the clinical characteristics in treatment-naive RA patients and treated RA patients. METHODS: RA patients who underwent cervical spine X-ray imaging from the Department of Rheumatology and Immunology of Peking University Third Hospital and Peking University Shenzhen Hospital from August 2015 to October 2019 and had clinical records of medication administration were included. Clinical and laboratory data including cervical symptoms and X-ray imaging data of cervical spine were collected. The constituent ratio of cervical spine instability in treatment-naive RA patients was statistically analyzed. The clinical data and laboratory data were analyzed by t-test, u-test and chi square to explore the clinical characteristics of the treatment-naive RA patients with cervical instability. RESULTS: Of the 408 RA patients, 105 patients were treatment-naive. Of the 105 treatment-naive patients, 82.9% (87/105) were female, with an average age of (52±14) years, the median duration of the disease was 24 months, the shortest history was 2 weeks, and the longest history was 30 years. 28.6% (30/105) of the treatment-naive RA patients showed cervical spine instability. The prevalence of cervical instability was 13.6% in the treatment-naive RA patients with disease duration less than 24 months. Among them, there were no significant differences in neck symptoms between cervical spine instability group and none cervical spine instability group. The patients with cervical spine instability had a longer duration of disease [60 (18, 180) months vs.16 (8, 51) months], a higher proportion of peripheral joint deformity (63.3%vs.21.3%), and a lower hemoglobin [(106.90±21.61) g/L vs. (115.77±14.69) g/L]. There was no significant difference in the occurrence of cervical instability in the treatment-naive RA patients compared with treated RA patients. Among the RA patients with cervical instability, there was no statistically significant difference in the composition of each type between the patients with treatment-naive RA and patients with treated RA, except for a shorter duration of disease [120.0 (72.0, 240.0) months vs. 60.0 (27.0, 167.5) months]. CONCLUSION: 28.6% of treatment-naive RA patients showed cervical spine instability. Cervical instability was also common in RA patients with a duration less than 24 months. There was no significant correlation between cervical instability and neck symptoms. Patients with cervical spine instability had a long-term disease, a higher proportion of peripheral joint deformity and a lower hemoglobin. Controlling the condition of RA early may help to control the progression of cervical involvement in patients with RA.

[Characteristics of human papillomavirus infection and abnormal cervical cytology in health check-up females in Shenzhen].

Objective: To describe the characteristics of human papillomavirus infection and thinprep cytologic test (TCT) outcome in health check-up females in Shenzhen. Methods: Use cross-sectional design, collect information from data from health check-up females in Shenzhen and describe characteristics of HPV infections screening and TCT outcomes. Results: We collected the data of 75 754 females, 103 508 females and 69 964 females received HPV detection, TCT and combined detection respectively. HPV standardized infection rate was 19.89% (95%CI: 19.45%-20.33%) and showed a "U-shaped" pattern in age distribution. The most prevalent HPV genotypes were 52, 51, 16, 58 and 53. Infection rate was higher for high-risk HPV than low-risk HPV genotype. Single infection was more common than its multiple infection. In addition, 7.48% (95%CI: 7.22%-7.75%) women were TCT positive, of whom 4.58% (95%CI: 4.40%-4.76%), 2.54% (2.40%-2.69%), 0.27% (95%CI: 0.23%-0.31%) had atypical squamous cells, low-grade squamous intraepithelial lesions and high-grade squamous intraepithelial lesions, respectively. Overall and subtype HPV infection rates increased with severity of abnormal cervical cytology. The most prevalent HPV genotypes were 52, 58 and 16 in women with abnormal cervical cytology. Conclusions: HPV prevalence remains at a high level in Shenzhen. This study suggests that attention should be paid to HPV screening, especially in young, perimenopausal women and in high risk HPV genotype infection. Timely follow-up and cervical cytology screening are required for women with high-risk HPV infection or persistent infection. Future vaccination strategies should take account of prevalent HPV genotype.

[Treatment strategy of gastrointestinal stromal tumors in the background of COVID-19].

COVID-19, caused by SARS-COV-2, has the characteristics of world epidemic, highly infectious and large base of death. In China, transmission route of SARS-COV-2 has been contained so effectively that COVID-19 has been well controlled due to the proactive national prevention and control strategy. However, not only does it bring a huge impact on the existing medical structure model, but also an objective impact on the treatment of patients with chronic diseases such as malignant tumors. Based on the progress reported in the domestic and international literatures and the actual management experience of our team, this paper reflects on the treatment strategies for patients with gastrointestinal stromal tumor (GIST) during the epidemic period of COVID-19. We focus on risk stratification for primary GIST and forming treatment strategies accordingly. Major considerations include the impact of delayed operation, the burden of medical resources, the waiting time for elective operation, and the principle of emergency operation. In addition, we focus on the level of evidence for non-surgical approaches with a view to developing a holistic strategy of "priority management principles" to guide clinical treatment in the context of limited resources and different GIST priorities.

[A rare anatomical variation of ileocolic veins involving in gastrocolic vein trunk: case report and literature review].

Objective: Due to its various anatomical variations and numerous branches, the gastrocolic vein trunk (Henle trunk) is the most common site to develop bleeding and other complications in laparoscopic right hemicolectomy for colon cancer. This study aims to investigate the role of ileocolic vein (ICV) joining with Henle trunk, a rare anatomical variation. Methods: A rare case whose ICV was newly found to involve in the formation of Henle trunk during laparoscopic resection of right hemicolon cancer was reported as right gastroepiploic vein+ right colic vein+superior right colic vein+ICV. This anatomical variation was confirmed by multi-slice spiral CT coronal two-dimensional reconstruction of right hemicolon angiography. The literatures about ICV participating in formation of Henle trunk were systematically searched from PubMed, The Cochran Library, CNKI net and Wanfang database, and the occurrence probability and composition of its anatomical variation were analyzed. Results: This was a 47-year-old female patient who underwent laparoscopic right hemicolectomy. When the vessels were dissected during operation, it was found that ICV did not accompany the ileocolic artery, but directly flowed into Henle trunk. Two-dimensional reconstructed CT images of right hemicolon vessels showed that the composition of Henle trunk was rarely varied, which was composed of right gastroepiploic vein, right colonic vein, superior right colonic vein and ICV. Five literatures were enrolled from literature retrieval. A total of 12 cases with ICV participating in the construction of Henle trunk were reported, with a probability of 0.27%-6.31% and 6 forms of the formation of Henle trunk. In this case, Henle trunk was made up of right gastroepiploic vein, right colonic vein, upper right colonic vein and ICV, which was reported for the first time. Conclusions: ICV involving in Henle trunk is a rare vascular variation, and this type of variation should be fully recognized. Careful dissection during operation is necessary to prevent intraoperative bleeding caused by improper operation.

[Anatomical controversies involved in radical resection of rectal cancer].

The concept of total mesorectal resection provides a quality control standard that can be followed for radical resection of rectal cancer, but some anatomical problems are still controversial. Compared with traditional open surgery, laparoscopic radical rectal surgery has better surgical vision, better neurological protection, better operating space. However, if the surgeon has insufficient understanding of the anatomy, collateral damage may occur, such as uncontrollable bleeding during the operation, postoperative urination and defecation dysfunction and so on. Based on the interpretation of the researches at home and abroad, combined with the clinical experience, we elucidate some associated issues, including anatomic variation of inferior mesenteric vessels, the controversy of inferior mesenteric artery ligation plane, the controversy of lymph node dissection in No. 253, the anatomical variation of middle rectal artery, and the anatomical controversy of lateral lymph node dissection in rectal cancer, in order to provide better cognitive process for the clinical front-line surgeons.

[Exploration on clinical treatment of incisional wound infection after orthopedic internal fixation for lower extremity joint injuries].

Objective: To explore the clinical treatment of incisional wound infection after orthopedic internal fixation for lower extremity joint injuries. Methods: A retrospective cohort study was conducted. From February 2014 to December 2019, 214 patients with lower limb closed injury were treated in Beijing Jishuitan Hospital, including 143 males and 71 females, aged from 16 to 65 years. All patients underwent orthopedic internal fixation, after which incisional wound infection developed in 42 cases of postoperative wounds of patellar fracture, 30 cases of postoperative wounds of tibial plateau fracture, 72 cases of postoperative wounds of Achilles tendon rupture, 45 cases of postoperative wounds of calcaneal fracture, and 25 cases of postoperative wounds of Pilon fracture with 31 cases of superficial infection and 183 cases of deep infection. According to the postoperative evaluation of the incisional wounds, dressing change and/or thorough debridement (with wound area from 4 cm×3 cm to 11 cm×5 cm after debridement), and internal fixation treatment were performed. After thorough debridement, wounds were treated with continuous vacuum sealing drainage (VSD), and then direct suture+VSD or flap transplantation were performed according to the incision and its surrounding skin tissue, infection, suture tension, exposure of internal fixation or bone or Achilles tendon tissue. According to the wound site and injury, the flap types of retrograde anterolateral thigh perforator flap, medial sural artery perforator flap, gastrocnemius myocutaneous flap, sural neurovascular flap, peroneal brevis muscle flap, posterior tibial artery perforator flap, and free anterolateral thigh perforator flap could be selected. The donor site wound was closed by direct suture or grafted with split-thickness skin graft. The removal of internal fixation, wound repair method, type and size of flaps, survival of flaps, and wound healing were recorded. The recurrence of infection, appearance of donor and recipient areas, and recovery of lower limb joint function were followed up. Results: All the internal fixations were removed in patients with postoperative wounds of patellar fracture, among whom 36 cases were repaired with retrograde anterolateral thigh perforator flap, and 6 cases were repaired with medial sural artery perforator flap. Among the patients with postoperative wounds of tibial plateau fracture, the internal fixation was completely retained in 18 cases, partially removed in 6 cases, and completely removed in 6 cases, while the wound was closed by direct suture in 8 cases, transplanted with gastrocnemius myocutaneous flap in 21 cases, and transplanted with medial sural artery perforator flap in 1 case. Among the patients with postoperative wounds of Achilles tendon rupture, the internal fixation was completely retained in 10 cases and completely removed in 62 cases, and the wound was closed by direct suture in 10 cases and transplanted with sural neurovascular flap in 62 cases. Among the patients with postoperative wounds of calcaneal fracture, the internal fixation was completely removed in 32 cases and completely retained in 13 cases, and the wound was healed by dressing change in 5 cases, closed by direct suture in 5 cases, transplanted with sural neurovascular flap in 23 cases, and transplanted with sural neurovascular flap combined with peroneal brevis muscle flap in 12 cases. Among the patients with postoperative wounds of Pilon fracture, the internal fixation was partially removed in 5 cases, completely retained in 17 cases, and completely removed in 3 cases; the wound was closed by direct suture in 4 cases, transplanted with posterior tibial artery perforator flap in 18 cases, and transplanted with free anterolateral thigh flap in 3 cases. The area of flaps/myocutaneous flaps ranged from 5 cm×3 cm to 18 cm×8 cm, and the area of muscle flaps were from 13.0 cm×1.5 cm to 15.0 cm×2.5 cm. All the wounds closed by direct suture healed. Blood flow obstacle occurred in the distal margin of sural neurovascular flap transplanted in 5 patients and posterior tibial artery perforator flap transplanted in one patient, which healed successfully after dressing change. The other flaps survived well, and the wounds were healed. The patients were followed up for 5 months to 5 years, and no recurrence of infection occurred. The direct suture of the flap donor site left linear scar, and the appearance of the skin graft was good. Three patients with tibial plateau fracture and 2 patients with Pilon fracture had limited joint movement, while the joint activity of the other patients was normal. The patients with Achilles tendon rupture and calcaneal fracture had normal ground motion. The sural neurovascular flap grafted on the wound after calcaneal fracture was bloated resulting in inconvenience in wearing shoes, the gastrocnemius myocutaneous flap grafted on the postoperative wound after tibial plateau fracture was bloated, whereas the appearance of the other flaps was good. Conclusions: For incisional wound infection after orthopedic internal fixation for lower extremity joint injuries, the treatment should be classified according to wound infection and wound site. Dressing change, thorough debridement, reasonable disposal of internal fixation, direct suture after application of VSD, and flap covering, etc. not only ensure the infection control and the wound closure, but also restore the function of the lower limbs to the greatest extent, so as to maximize the benefit of the patient.

MiRNA-15a-3p inhibits the metastasis of hepatocellular carcinoma by interacting with HMOX1.

OBJECTIVE: To uncover the relationship between microRNA-15a-3p (miRNA-15a-3p) level and clinical features of hepatocellular carcinoma (HCC), and to explore the influence of miRNA-15a-3p on metastasis of HCC cells. PATIENTS AND METHODS: HCC and paracancerous tissues were surgically resected from 44 HCC patients. Their clinical data and follow-up files were recorded. Differential expressions of miRNA-15a-3p in HCC samples were determined. The relationship between miRNA-15a-3p level and clinical features of HCC patients was analyzed. Changes in proliferative, migratory and invasive potentials in Huh7 and HepG2 cells overexpressing miRNA-15a-3p were examined. The downstream gene of miRNA-15a-3p and its involvement in HCC development were finally explored. RESULTS: MiRNA-15a-3p was downregulated in HCC tissues. High metastasis rate and poor prognosis were observed in HCC patients expressing a low level of miRNA-15a-3p. Overexpression of miRNA-15a-3p attenuated proliferative, migratory and invasive potentials in HCC. Protein levels of HMOX1, CD31, c-Myc, MMP-2 and MMP-9 were downregulated in HCC cells after overexpression of miRNA-15a-3p. HMOX1 was the downstream gene of miRNA-15a-3p, which was upregulated in HCC samples. Highly expressed HMOX1 was unfavorable to the prognosis in HCC. Overexpression of HMOX1 abolished the regulatory effects of miRNA-15a-3p on HCC cell phenotypes. CONCLUSIONS: MiRNA-15a-3p is closely linked to lymphatic metastasis, distant metastasis and poor prognosis in HCC. It inhibits the malignant development of HCC by interacting with HMOX1.

[Population distribution and clinical characteristics in rheumatoid arthritis patients with cervical spine instability].

OBJECTIVE: To investigate the population distribution of cervical spine instability in rheumatoid arthritis (RA) patients, and to analyze the clinical characteristics in RA patients with cervical spine instability. METHODS: A total of 439 RA patients who had completed cervical spine X-ray examination from Department of Rheumatology and Immunology of Peking University Shenzhen Hospital and Peking University Third Hospital from August 2015 to March 2019 were enrolled. The clinical data, laboratory data and cervical radiographic data were collected and analyzed by t-test, rank sum test and Chi-square test to clarify the clinical characteristics in the RA patients with cervical spine instability. RESULTS: Of the 439 RA patients, 80.9% (355/439) were female, with an average age of (52.9±13.9) years, a median duration of the disease was 60 months, the shortest history was 2 weeks, and the longest history was up to 46 years. 29.6% (130/439) of the RA patients showed cervical spine instability. Among them, 20 RA patients were complicated with two different types of cervical instability, the atlantoaxial subluxation (AAS) accounted for 24.6% (108/439), the vertical subluxation (VS) accounted for 7.3% (32/439) and the subluxial subluxations (SAS) accounted for 2.3% (10/439). The patients with cervical spine instability had a longer duration of disease [120 (36, 240) months vs. 48 (12, 120) months], a higher proportion of peripheral joint deformity (56.9% vs. 29.9%), and a higher visual analog scale (VAS) measuring general health score (4.89±2.49 vs. 3.93±2.38), a lower hemoglobin [(111.31±19.44) g/L vs. (115.56±16.60) g/L] and a higher positive rate of anti-cyclic citrullina-ted peptide (CCP) antibody (90.8% vs. 76.6%). There were no significant differences in gender, age, number of swollen joints, number of tenderness joints, erythrocyte sedimentation rate, rheumatoid factor level, 28-joint disease activity score, positive rate of anti keratin antibody, duration of glucocorticoid use and duration of disease modifying anti-rheumatic drugs use between the two groups. CONCLUSION: In the study, 29.6% of the RA patients showed cervical spine instability. RA patients with cervical spine instability had a long-term disease, a higher proportion of peripheral joint deformity, a higher VAS measuring general health score, a lower hemoglobin and a higher positive rate of anti-CCP antibody.

[Mechanism study of dendritic epidermal T lymphocytes in promoting healing of full-thickness skin defects wound on mice by regulating the proliferation and differentiation of epidermal stem cells in mice].

Objective: To explore the mechanism of dendritic epidermal T lymphocytes (DETCs) in promoting healing of full-thickness skin defect wound on mice by regulating the proliferation and differentiation of epidermal stem cells (ESCs) in mice. Methods: (1) Ten 8-week-old wild type (WT) male C57BL/6 mice (the same sex and kind below) were sacrificed to collect the skin of back for extracting DETCs to culture. Five WT and five 8-week-old T cell receptor (TCR) δ(-)/(-) mice were selected and enrolled in WT control group and TCR δ(-)/(-) control group, respectively. A full-thickness skin defect wound with diameter of 6 mm was made on both sides of spinal line on the back of mice without any treatment after injury. Another fifteen 8-week-old TCR δ(-)/(-) mice were selected and divided into phosphate buffer solution (PBS), DETC, and insulin-like growth factor-Ⅰ(IGF-Ⅰ) groups according to the random number table (the same grouping method below), with 5 mice in each group, and the same full-thickness skin defect wound was made on each mouse. Immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 µL sterile PBS , DETCs (cell concentration of 1×10(6)/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. The percentage of the residual wound area was calculated on post injury day (PID) 2, 4, 6, and 8. (2) Three 8-week-old WT mice were enrolled in WT control group and nine 8-week-old TCR δ(-)/(-) mice were divided into TCR δ(-)/(-) control group, PBS group, and DETC group, with 3 mice in each group. The full-thickness skin defect wound was made as in experiment (1) . On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin was detected by chemiluminescence imaging analyzer. (3) Three 8-week-old WT mice were enrolled in WT control group and six 8-week-old TCR δ(-)/(-) mice were divided into PBS and DETC groups, with 3 mice in each group, and the full-thickness skin defect wound was made as in experiment (1). On PID3, DETCs were extracted from the wound margin epidermis tissue to detect the percentage of DETCs expressing IGF-Ⅰ by flow cytometer. (4) The mice were taken as in experiment (2) and divided into WT control, PBS, DETC, and IGF-Ⅰ groups. A straight full-thickness skin defect incision with length of 3 cm was made in the direction of one inner ear. Mice in WT control group didn't have any other treatment after injury, and immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 µL sterile PBS, DETCs (cell concentration of 1×10(6)/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. On PID 12, epidermis tissue of wound margin was collected, and immunofluorescence staining was performed to observe the number of keratin 15 positive cells. (5) The same mice were collected, grouped, and treated as in experiment (4). On PID12, the epidermis tissue of wound margin was collected and immunofluorescence staining was performed to observe the number of keratin 10 positive cells. (6) Twenty 3-day-old WT mice (the same below) were sacrificed to collect the whole skin, which was used to extract ESCs, with 5 mice detecting one index. The ESCs were divided into DETC co-culture group and control group, which were added with 1 mL DETCs (cell concentration of 1.25×10(6)/mL) and DETC medium, respectively. The percentage of 5-ethynyl-2'-deoxyuridine (EdU) positive cell on culture day (CD) 3, the percentages of CD49f(+) CD71(-) and keratin 14 positive cells on CD 5, and the percentage of keratin 10 positive cell on CD 10 in 2 groups were detected by flow cytometer. (7) Twenty mice were taken to extract ESCs, with 5 mice detecting one index. The ESCs were divided into control group and IGF-Ⅰ group, which were added with 1 mL sterile PBS and 10 ng/mL recombinant mice IGF-Ⅰ, respectively. The percentages of EdU positive cell, CD49f(+) CD71(-) cell, keratin10 positive cell, and keratin 14 positive cell were detected as in experiment (6). The sample in each group of experiments (6) and (7) was three. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and t test. Results: (1) On PID 4, 6, and 8, the percentage of residual wound area in TCR δ(-)/(-) control group was significantly higher than that in WT control group (t=2.78, 3.39, 3.66, P<0.05 or P<0.01). The percentage of residual wound area in DETC group and IGF-Ⅰgroup on PID 4, 6, and 8 was apparently lower than that in PBS group (t=2.61, 3.21, 3.88, 2.84, 2.91, 2.49, P<0.05 or P<0.01). (2) On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in TCR δ(-)/(-) control group was significantly lower than that in WT control group (t=17.34, P<0.01). The protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in DETC group was significantly higher than that in PBS group (t=11.71, P<0.01). (3) On PID 3, the percentage of DETCs expressing IGF-Ⅰ in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group and DETC group (t=24.95, 27.23, P<0.01). (4) On PID 12, the number of keratin 15 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group, DETC group, and IGF-Ⅰ group (t=17.97, 11.95, 7.63, P<0.01). (5) The number of keratin 10 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly higher than that in WT control group, DETC group, and IGF-Ⅰ group (t=11.59, 9.51, 3.48, P<0.05 or P<0.01). (6) The percentages of EdU positive cells on CD 3, CD49f(+) CD71(-) cells on CD 5, and keratin 14 positive cells on CD 5 in DETC co-culture group were respectively (43.5±0.6)%, (66.5±0.5)%, (69.3±1.7)%, apparently higher than (32.3±1.3)%, (56.4±0.3)%, (54.9±1.3)% in control group (t=7.97, 17.10, 6.66, P<0.01). The percentage of keratin 10 positive cells on CD 10 in DETC co-culture group was (55.7±0.7)%, significantly lower than (67.1±1.2)% in control group (t=8.34, P<0.01). (7) The percentages of EdU positive cells on CD 3, CD49f(+) CD71(-) cells on CD 5, and keratin 14 positive cells on CD 5 in IGF-Ⅰ group were respectively (42.1±0.9)%, (81.1±1.3)%, (66.8±1.0)%, apparently higher than (32.4±0.7)%, (74.9±0.7)%, (52.0±1.9)% in control group (t=8.39, 4.24, 7.25, P<0.05 or P<0.01). The percentage of keratin 10 positive cells on CD 10 in IGF-Ⅰ group was (53.5±1.1)% , significantly lower than (58.2±0.3)% in control group (t=3.99, P<0.05). Conclusions: DETCs can promote the proliferation and anti-apoptotic potential of ESCs and inhibit their differentiation into end-stage by secreting IGF-Ⅰ, thus promoting wound healing of full-thickness skin defects in mice.

[Study on mechanisms of interleukin-17A regulating the expressions of interleukin-1ß and interleukin-23 in mouse keratinocytes].

Objective: To investigate the mechanisms of interleukin-17A (IL-17A) regulating the expressions of IL-1ß and IL-23 in mouse keratinocytes (KCs). Methods: Primary KCs were isolated from the skin of 400 newborn male and female wild type C57BL/6 mice and cultured in 24-well plates with Roswell Park Memorial Institute 1640 medium containing fetal bovine serum in the volume fraction of 10% for the following experiments. (1) The cells were divided into phosphate buffer solution (PBS) control group and IL-17A stimulation group according to the random number table (the same grouping method below), which were cultured with 10 µL PBS or 10 µL IL-17A in the mass concentration of 100 ng/mL for 6 hours, respectively. The expression levels of IL-1ß and IL-23 mRNA in cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with 3 samples in each group. (2) The cells were divided into dimethyl sulfoxide (DMSO) control group, IL-17A+ DMSO group, IL-17A+ nuclear factor κB (NF-κB) inhibitor group, IL-17A+ signal transduction and activator of transcription 3 (STAT3) inhibitor group, IL-17A+ extracellular signal-regulated kinase 1 (ERK1) inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ c-Jun N-terminal kinase (JNK) inhibitor group. The reagents were added to cells in corresponding groups respectively and cultured for 6 hours. The volume of each reagent was 10 µL, the mass concentration of IL-17A was 100 ng/mL, and the molarity concentrations of NF-κB, STAT3, ERK1, ERK2, JNK signal pathway inhibitors PDTC, S3I-201, SCH772984, SCH772984, SP600125 were 5 µmol/L, 100 µmol/L, 4 nmol/L, 1 nmol/L, and 10 µmol/L, respectively. The expression levels of IL-1ß mRNA and IL-23 mRNA in cells were detected by real-time fluorescence quantitative RT-PCR, with 3 samples in each group. (3) The cells were grouped and treated the same as those in experiment (1). The levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation were detected by Western blotting, with 3 samples in each group. Data were statistically analyzed with two-tailed Student t test, one-way analysis of variance, t test, and Bonferroni correction. Results: (1) After culture of 6 hours, compared with those in PBS control group, the expression levels of IL-1ß and IL-23 mRNA in cells in IL-17A stimulation group were significantly increased (t=13.46, 6.72, P<0.01). (2) After culture of 6 hours, the expression levels of IL-1ß and IL-23 mRNA in cells in DMSO control group, IL-17A+ DMSO group, IL-17A+ NF-κB inhibitor group, IL-17A+ STAT3 inhibitor group, IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group were 1.00±0.11, 4.01±0.32, 0.32±0.06, 1.76±0.43, 3.62±0.24, 3.80±0.43, 4.26±0.74 and 1.03±0.29, 4.08±0.34, 4.76±0.38, 4.70±0.21, 1.06±0.42, 0.92±0.21, 0.39±0.05, respectively. Compared with those in DMSO control group, the expression levels of IL-1ß and IL-23 mRNA in cells in IL-17A+ DMSO group were significantly increased (t=9.24, 12.60, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-1ß mRNA was significantly decreased in cells in IL-17A+ NF-κB inhibitor group and IL-17A+ STAT3 inhibitor group (t=11.34, 6.91, P<0.01). Compared with that in IL-17A+ DMSO group, the expression level of IL-23 mRNA was significantly decreased in cells in IL-17A+ ERK1 inhibitor group, IL-17A+ ERK2 inhibitor group, and IL-17A+ JNK inhibitor group (t=12.44, 13.03, 15.21, P<0.01). (3) After culture of 6 hours, compared with those in PBS control group, the levels of NF-κB phosphorylation, STAT3 phosphorylation, ERK phosphorylation, and JNK phosphorylation in cells in IL-17A stimulation group were significantly increased. Conclusions: IL-17A promotes the transcription of IL-1ß in mouse KCs through the phosphorylation of NF-κB and STAT3 pathways and IL-23 through the phosphorylation of ERK and JNK pathways.

Lipoxin A4 alleviates lung injury in sepsis rats through p38/MAPK signaling pathway.

The aim of the study was to explore the effect of lipoxin A4 (LXA4) on lung injury in sepsis rats through the p38/mitogen-activated protein kinase (MAPK) signaling pathway. Sprague-Dawley rats were used for the study. The rat model of sepsis-induced acute lung injury was established via cecal ligation (Sepsis group, n=20). LXA4 (0.1 mg/kg) was injected at 6 h after modeling (Treatment group, n=20), and a The Control group (n=20) was also set up. The 7-day survival rate was 100% in The Control group, and LXA4 raised the survival rate of rats in the Sepsis group from 40% to 60% (P<0.01). Alveolar fluid clearance (AFC) significantly declined and the wet/dry weight (W/D) ratio of lung tissues rose remarkably in the Sepsis group compared with those in the Control group, while LXA4 restored AFC and reduced the W/D ratio of lung tissues (P<0.05), suggesting that LXA4 treatment reduces lung fluids and partially enhances AFC, thus lowering the W/D ratio of lung. The total cell count, polymorphonuclear neutrophils (PMN) percentage and concentration of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in bronchoalveolar lavage fluid (BALF) were obviously increased in the Sepsis group compared with those in the Control group, while they were markedly decreased in the Treatment group (P<0.05). The activity of myeloperoxidase (MPO) in lung tissue homogenate was evidently higher in the Sepsis group than that in The Control group, while it was notably lower in the Treatment group than that in the Sepsis group after LXA4 treatment (P<0.05). Moreover, it was observed microscopically that the morphology of lung tissues was intact in the Control group. Finally, the results of Western blotting manifested that the p-p38/ MAPK protein expression was remarkably increased in the Sepsis group, indicating the activation of the p38/MAPK pathway, while it was remarkably decreased in the Treatment group, indicating the inhibited activity of the pathway (P<0.05). LXA4 has an anti-inflammatory effect on sepsis rats with lung injury, and such effect is related to the p38/MAPK signaling pathway.

[Epidemiological investigation of 169 inpatients with high-voltage electrical burns on the wrist].

Objective: To analyze the epidemiological characteristics of inpatients with high-voltage electrical burns on the wrist in Beijing Jishuitan Hospital (hereinafter referred to as the author's unit), so as to provide reference for the prevention and treatment of high-voltage electrical burns on the wrist. Methods: The medical records of inpatients suffered from high-voltage electrical burns on the wrist in the author's unit from January 2008 to December 2019 were collected. The patients' gender, age, population category, injury situation, injury season, total burn area, electrical burn type on the wrist of the affected limbs, the time from injury to first flap/myocutaneous flap transplantation, vascular reconstruction and flap/myocutaneous flap transplantation of the affected limbs, the total amputation rate of the affected limbs and the amputation rate of the affected limbs with type Ⅲelectrical burns on the wrist, the number of operation, postoperative infection rate of flap/myocutaneous flap, length of hospital stay, hospitalization expense, and treatment outcome were retrospectively analyzed. Comparison of the aforementioned statistical items between patients admitted from January 2008 to December 2013 (hereinafter referred to as the pre-stage) and January 2014 to December 2019 (hereinafter referred to as the post-stage) except gender, the total amputation rate of the affected limbs, treatment outcome were performed. Data were statistically analyzed with chi-square test, Fisher's exact probability test, and Wilcoxon rank-sum test. Results: During the 12 years, a total of 169 patients with high-voltage electrical burns on the wrist were admitted to the author's unit, including 162 males and 7 females, aged (35±13) years, and 75.15% (127/169) of patients were 21-50 years old. The top three groups in population category from high to low were workers, migrant workers, and primary and secondary school students, accounting for 48.52% (82/169), 28.99% (49/169), and 9.47% (16/169), respectively. At work of non-electric power accounted for 47.93% (81/169) and ranked the first in the proportion of injury situation. The injury occurred mostly in summer and autumn, accounting for 39.05% (66/169) and 28.99% (49/169), respectively. About 65.09% (110/169) of the patients were with total burn area less than 10% total body surface area. There were totally 216 affected limbs with high-voltage electrical burns on the wrist, of which the numbers of wrist with type Ⅲ and type Ⅳ injury were 25 (11.57%) and 21 (9.72%), respectively. The time from injury to first flap/myocutaneous flap transplantation was 6.00 (3.75, 8.00) d. There were 45 affected limbs operated with vascular reconstruction, 75 affected limbs transplanted with pedicled axial flap, and 86 affected limbs transplanted with free flap/myocutaneous flap. The total amputation rate of affected limbs was 12.96% (28/216), and the amputation rate of the affected limbs with type Ⅲ electrical burns on the wrist was 28% (7/25). The number of operation was 4 (3, 5) times, the postoperative infection rate of flap/myocutaneous flap was 7.18% (13/181), the hospitalization time was 39.00 (25.00, 50.00) d, and the hospitalization cost was 123 553.00 (50 656.50, 216 003.00) yuan. Compared with those of the pre-stage, the time from injury to first flap/myocutaneous flap transplantation was significantly shortened (Z=-4.038, P<0.01), the ratio of free flap/myocutaneous flap transplantation on affected limbs was significantly increased (χ(2)=13.478, P<0.01), the ratio of pedicled axial flap transplantation on affected limbs was significantly decreased (χ(2)=10.242, P<0.01), the number of operation was significantly reduced (Z=-5.903, P<0.01), the postoperative infection rate of flap/myocutaneous flap was significantly decreased (χ(2)=4.492, P<0.05), the length of hospital stay was significantly shortened (Z=-2.723, P<0.01), and the hospitalization expense was significantly decreased among patients hospitalized in the post-stage (Z=-2.121, P<0.05). The other items were close between patients hospitalized in the pre-stage and those in the post-stage. Conclusions: Men were more likely than women to suffer from high-voltage electrical burns on the wrist in the author's unit. Young workers and migrant workers may be the key groups for the prevention of high-voltage electrical burns on the wrist. It is very important to strengthen the management of high voltage safety education for non-electric power staff, warn and protect the surrounding area of high voltage environment, and prohibit non staff for entering the high voltage environment, especially in summer and autumn. In the last 6 years, the patients with high-voltage electrical burns on the wrist may benefit from the active vascular reconstruction at early stage and free flap/myocutaneous flap repair in the author's unit.