Background Studies confirmed that hydroxycamptothecin cause the apoptosis of human Tenon capsule fibroblasts (HTFs) by protein kinase R-like endoplasmic reticulum stress kinase (PERK) single pathway.Autophagy and apoptosis are programmed cell death following stress reaction,so they remain a close association.However,the effect of hydroxycamptothecin on the autophagy of HTFs and its mechanism are still unclear.Objective This study was to explore the promoting effect of PERK signal pathway on hydroxycamptothecin inducing the autophagy of HTFs.Methods This study procedure was approval by Ethic Committee of Nanjing Medical University.Human Tenon capsule tissue was obtained from fresh adult donors.HTFs were cultured and passaged by explant-culture method and identified by immunofluorescence for vimentin and keratin.pLVX-PERK lentiviral packed by 293T cells was transfected into HTFs to obtain stable PERK-knockout cell line by puromycin selection.Then the HTFs were treated with 0.10 g/L of hydroxycamptothecin for 5 minutes and consecutively cultivated for 24 hours,and the untreated cells were used as the control group.Western blot assay was used to detect the expressions of autophagy specific proteins in the cells,including autophagy related gene 5 (ATG-5),Beclin-1,light chain 3 (LC-3).Cyto-ID staining was used to identify the autophagosome in the cells.The experimental results were analyzed and compared between different treating groups.Results The gray scales for the expressions of Beclin 1,ATG-5,LC-3-Ⅰ and LC-3-Ⅱ proteins in HTFs were 0.365:±0.045,0.765 ±0.055,0.120±0.030 and 0.215 ±0.035 in the control group,and those in the hydroxycamptothecin treated group were 0.980±0.070,1.495±0.095,0.585±0.025 and 0.785±0.055,showing a significant decline in the hydroxycamptothecin treated group(P=0.018,0.022,0.007,0.013).The green fluorescence of the autophagosome was stronger in the hydroxycamptothecin treated group compared with the control group.Western blot revealed that the gray scale of PERK expression in the cells was 0.130±0.030 in the PERK-knockout group,with a significant reduce in comparison with 0.765 ±0.055 of the control group (P =0.010).However,no obvious distinctions were seen in the band intensities of the expressions of Beclin-1,ATG-5 and LC-3 proteins between the two groups.Western blot indicated that the grey scale of the PERK expression in the cells was 1.790± 0.060 in the 0.10 g/L hydroxycamptothecin group,which was significantly higher than 0.880 ± 0.070 of the control group (P =0.010).Expression levels (gray scales) of Beclin-1,ATG-5,LC-3-Ⅰand LC-3-Ⅱ in the PERK-knockout+ 0.10 g/L hydroxycamptothecin group were 0.475 ± 0.045,0.390 ± 0.040,0.055 ± 0.015 and 0.075 ± 0.025,which were significantly lowed in comparison with 0.955 ± 0.065,0.765 ± 0.055,0.155 ± 0.015 and 0.280 ± 0.030 of the control+ 0.10 g/L hydroxycamptothecin group (P =0.026,0.031,0.042,0.034).In addition,the fluorescence intensity of autophagosomes was weaker in the PERK-knockout+0.10 g/L hydroxycamptothecin group compared with the control+0.10 g/L hydroxycamptothecin group.Conclusions Hydroxycamptothecin induces the autophagy of HTFs by PERK signal pathway.

Objective To improve the method of blood collection from external jugular vein in rats,so as to repeatedly obtain blood samples of high quality and to meet the special requirements of the experiment.Methods The experimental rats were anesthetized by intraperitoneal anesthesia,and the jugular vein was fully exposed with special fixed method.Whereafter,the blood samples were collected by disposable vacuum blood vessels.Results This method had little traumatic effect.The blood samples were collected from 60 rats at an interval of 14 days with 1.8 to 2.5 mL each time.The success rate was above 95％,and the normal activities of the rats were not apparently affected.This vacuum blood collection method guaranteed the quality of serum,and the veracity of blood detection and analysis were also significantly improved.Conclusion On the basis of the principle of animal welfare,the improved rat jugular vein blood collection method has the advantages of simple operation,less trauma,large blood collection,high blood sample quality aod so on,which can ensure the effect of the special experiment.

Objective: To establish a mouse model of acute lung injury (ALI) via intratracheal aerosolization of paraquat (PQ). Methods: Thirty male C57BL/6J mice of 8-10 weeks old were randomly(random number) divided into the paraquat model group (n = 15) and the saline control group (n = 15). Mice in the paraquat model group were administered PQ solution (0.4 mg/mL) at a single dose of 50 μL per mouse via intratracheal aerosolization. Mice in the saline control group received an equal volume of saline. Mice were sacrificed 72 h post-PQ administration and samples were collected. The general physical condition of mice including body weight as well as mortality was monitored daily. Pulmonary function, cell counting and classification of bronchoalveolar lavage fluid (BALF), proinflammatory cytokines (TNF-α, IL-1β, and IL-6) levels of BALF and serum, MPO activity and total protein concentration of BALF and pathological lung injury scores were obtained. Results: The body mass of mice decreased progressively within 72 h after PQ intratracheal administration. The static compliance of lung decreased (P<0.01) and the resistance increased (P<0.01). The number of inflammatory cells in BALF was significantly higher than that of the saline control group (all P<0.01), the levels of proinflammatory cytokines were significantly upregulated in both BALF and serum (all P<0.01), and the activity of MPO in BALF was also remarkedly increased (P<0.01). The total protein concentration of BALF was significantly elevated (P<0.01). The pathological observation of lung tissue showed a large number of inflammatory cells infiltration accompanying pulmonary hemorrhage, alveolar collapse, hyaline membrane formation and alveolar septal thickening, and the lung injury scores significantly increased (P<0.01). Conclusions Intratracheal aerosolization of a single dose of 0.02 mg PQ can successfully induce ALI in mice.

Objective:To investigate the application value of computed tomography (CT) and magnetic resonance imaging (MRI) examination in clinical diagnosis of gallbladder tumor with perigallbladder invasion.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 80 gallbladder tumor patients with perigallbladder invasion who were admitted to 3 medical centers (21 cases in The First Affiliated Hospital of Northwest University, 42 cases in The First Affiliated Hospital of the Air Force Medical University, 17 cases in The First Affiliated Hospital of Dalian Medical University) from January 2021 to December 2022 were collec-ted. There were 45 males and 35 females, aged (56±4)years. Observation indicators: (1) CT and MRI examinations; (2) surgical conditions. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) CT and MRI examinations. Of 80 patients, cases with gallbladder cancer and gallbladder adenoma were 73 and 7, respectively. Cases with endoluminal nodular type, mass type and localized thick-walled type were 33, 39 and 8, respectively, with tumor diameter as 1.55 cm×1.35 cm×1.33 cm, 1.64 cm×1.37 cm×1.36 cm and 5.72 cm×4.07 cm×4.36 cm. Results of CT examination of endoluminal nodular type showed local nodular protrusions into the endoluminal area, and local enhancement on enhanced scanning. Results of CT examination of localized thick-walled type showed the cavity wall of lesion was locally or diffusely irregul-arly thickened, with a thickness of 1.10(range, 1.10-2.21)cm. Of 80 patients, results of CT and MRI examinations showed invasion of liver parenchyma in 68 cases, which was manifested as local mass, blurred demarcation, and abnormal protrusion. The maximum depth was (4.22±0.25)cm, (4.22±0.22)cm, (4.28±0.16)cm of cross-sectional, coronal, sagittal view in CT examination, respectively. The minimum depth was (0.22±0.10)cm, (0.25±0.08)cm, (0.24±0.12)cm. The depth of liver parenchyma invaded was (1.64±1.38)cm, (1.68±1.46)cm, (1.66±1.40)cm. Results of CT and MRI examinations showed invasion of perigallbladder, which was manifested as local invasion of the gastric antrum in 12 cases. (2) Surgical conditions. Of 80 patients, results of CT and MRI examina-tions showed that 60 patients had localized masses in the gallbladder cavity with or without infiltration of surrounding tissues. After confirming the absence of other organs and distant metastasis, cases undergoing radical resection and palliative resection were 44 and 16, respectively. Results of CT and MRI examina-tions showed that 20 patients had malignant gallbladder tumors with peri-pheral liver infiltration and multiple intrahepatic metastases with distant organ metastases, which were unresectable.Conclusion:For patients with gallbladder cancer and perigallbladder invasion, CT or MRI examina-tions can show their structural characteristics.

Background: and Purpose Collateral circulation is considered an important factor affecting the risk of stroke, but the factors that affect collateral circulation remain unclear. This study was performed to identify the factors associated with collateral circulation, especially blood lipids. Methods: The study involved patients who had undergone digital subtraction angiography and were confirmed as having severe unilateral stenosis or occlusion of the internal carotid artery (ICA). We classified the collateral circulation status of each patient as good (Grade 3 or 4) or poor (Grade 0, 1, or 2) according to the grading system of the American Society of Interventional and Therapeutic Neuroradiology/American Society of Interventional Radiology. We collected data on patients’ characteristics and identified the factors that affect collateral circulation. Results: This study included 212 patients. The multivariate logistic regression analysis showed that the high-density lipoprotein cholesterol (HDL-C) concentration and a complete anterior half of the circle of Willis were independent protective factors for good collateral circulation, whereas elevated lipoprotein(a) [Lp(a)] and serum creatinine concentrations were independent risk factors for good collateral circulation. The area under the receiver operating characteristics curve (AUC) was 0.68 (95% confidence interval [CI], 0.61–0.76) for HDL-C and 0.69 (95% CI, 0.62–0.76) for Lp(a). A binary logistic regression model analysis of the joint factor of HDL-C and Lp(a) yielded an AUC of 0.77 (95% CI, 0.71–0.84). Conclusions In patients with severe unilateral ICA stenosis or occlusion, the combination of HDL-C and Lp(a) is a useful predictor of collateral circulation.

Objective:To investigate and compare the regulatory effects of umbilical cord mesenchymal stem cells (MSC) and their conditioned medium (MSC-CM) on gut microbiota of septic mice.Methods:Twenty-eight six-to-eight-week-old female C57BL/6J mice were randomly divided into sham operation group (Sham group), sepsis model group (CLP group), sepsis+MSC treatment group (CLP+MSC group) and sepsis+MSC-CM treatment group (CLP+MSC-CM group), with seven mice in each group. The septic mouse model was established by cecal ligation and puncture (CLP). In Sham group, CLP were not performed, and other operations were the same as CLP group. Mice in the CLP+MSC group and CLP+MSC-CM group received 0.2 mL 1×10 6 MSC or 0.2 mL concentrated MSC-CM via intraperitoneal injection 6 hours after CLP, respectively. Sham group and CLP group were given 0.2 mL sterile phosphate buffer saline (PBS) via intraperitoneal injection. Histopathological changes were evaluated by hematoxylin-eosin (HE) staining and colon length. Levels of inflammatory factors in serum were detected by enzyme-linked immunosorbent assay (ELISA). Phenotype of peritoneal macrophages was analyzed by flow cytometry, and the gut microbiota was analyzed via 16S rRNA sequencing. Results:Compared with Sham group, significant inflammatory injury in lung and colon was observed, and shorter colon was detected in CLP group (cm: 6.00±0.26 vs. 7.11±0.09), the level of inflammatory cytokine interleukin-1β (IL-1β) in serum was significantly increased (ng/L: 432.70±17.68 vs. 353.70±17.01), the proportion of F4/80 + peritoneal macrophages was increased [(68.25±3.41)% vs. (50.84±4.98)%], while the ratio of F4/80 +CD206 + anti-inflammatory peritoneal macrophages was decreased [(45.25±6.75)% vs. (66.66±3.36)%]. The α diversity sobs index of gut microbiota was downregulated significantly (118.50±23.25 vs. 255.70±6.87), the structure of species composition was altered, and the relative abundance of functional gut microbiota related to transcription, secondary metabolites biosynthesis, transport and catabolism, carbohydrate transport and metabolism, and signal transduction were decreased significantly in CLP group (all P < 0.05). Compared with CLP group, upon MSC or MSC-CM treatment, the pathological injury in lung and colon was alleviated to varying extent, the length of colon was increased (cm: 6.53±0.27, 6.87±0.18 vs. 6.00±0.26), the level of IL-1β in serum was downregulated (ng/L: 382.10±16.93, 343.20±23.61 vs. 432.70±17.68), the ratio of F4/80 + peritoneal macrophages was decreased [(47.65±3.93)%, (48.68±2.51)% vs. (68.25±3.41)%], the ratio of F4/80 +CD206 + anti-inflammatory peritoneal macrophages was increased [(52.73±5.02)%, (66.38±4.73)% vs. (45.25±6.75)%], and the α diversity sobs index of gut microbiota was increased (182.50±16.35, 214.00±31.18 vs. 118.50±23.25), and the effects of MSC-CM were more significant (all P < 0.05). At the same time, species composition of gut microbiota was rebuilt, and a tendency of increase in relative abundance of functional gut microbiota was observed upon MSC and MSC-CM treatment. Conclusion:Both MSC and MSC-CM could alleviate inflammatory injury in tissues, and showed regulatory effects on gut microbiota in septic mouse model, moreover, MSC-CM exhibited superior advantages over MSC.

Objective:To investigate the role of TiRobot combined with O-arm assisted closed reduction and percutaneous screw fixation in unstable pelvic fracture surgery.Methods:Twelve patients with unstable pelvic fractures, 7 males and 5 females, aged 39.41±12.56 years (range 25-60 years), admitted from January 2020 to January 2021, were retrospectively analyzed. The duration between injury and surgery was 9.67±5.81 d (range 4-24 d). The following are the causes of injury, namely traffic injury in 6 cases, fall from height injury in 4 cases, crush injury in 2 cases. Tile type of pelvic fractures were 3 cases of type B (2 cases of type B2, 1 case of type B3), 9 cases of type C (4 cases of type C1, 3 cases of type C2, 2 cases of type C3). Sacral fractures with Denis division were 5 cases of zone I, 7 cases of zone II. Seven patients were repositioned by intraoperative traction on the injured side of the limb and maintained by fixation with a Starr frame. Two cases were repositioned by intraoperative longitudinal bone traction on each lower limb for fracture displacement. The other 3 B-type fractures were repositioned by inserting Schanz nails into the anterior inferior iliac spine bilaterally. Then, the fractures were maintained with external pelvic fixators. The fractures were assessed by O-arm fluoroscopy. Hollow screws were placed with the aid of orthopedic robotic guidance, and the screw positions were verified by O-arm fluoroscopy again after the nail placement. The duration of each screw placement and operation were recorded. The quality of fracture repositioning, hip function and postoperative complications were observed at the follow-up.Results:All patients were followed up for 8.17±1.99 months (range, 6-12 months). There were 27 screws inserted in 12 patients, with a single screw insertion duration of 19.9±1.8 min (range, 9-31 min). In all patients, one O-arm fluoroscopy was performed in each row after closed reduction and after completion of nail placement. The operative duration was 257.78±80.63 min (range, 160-450 min). The O-arm fluoroscopy after nail placement verified that 23 screws were placed in a satisfactory position at one time, with satisfactory rate of 85% (23/27) for the first nail placement. Two patients with preoperative grade II sacral plexus nerve injury recovered to grade I in one case and grade II in one case. However, the numbness was reduced after operation. No further aggravation of nerve injury occurred after surgery. The quality of fracture repositioning was evaluated according to the Matta score, including excellent in 9 cases, good in 2 cases, and acceptable in 1 case, with an excellent rate of 92% (11/12). At the last follow-up, the Majeed pelvic fracture quantitative assessment system scored 85.75±5.82 (range, 74-96). There were 8 cases in excellent and 4 cases in good. Twelve patients had one-stage wound healing. No complications, such as incision infection, nerve injury, loosening and breaking of the internal fixation occurred during the follow-up.Conclusion:TiRobot combined with O-arm assisted closed reduction and percutaneous screw fixation for unstable pelvic fractures has some advantages, including safety, precision, convenient nail placement, and less fluoroscopic radiation.

The new business forms are the new patterns of the job market extended by the use of mobile internet, big data, cloud computing, and other information technologies in the context of a new round of information technology development. In recent years, under the catalysis of the new employment model, a large number of new business employees have emerged such as food delivery riders, couriers, and online car-hailing drivers, whose employment forms are flexible and don’t fully conform to the establishment of labor relations with the employers, and whose problem of overtime work is common. The employment characteristics of new business forms of "focusing on performance and ignoring responsibility" make workers at great risks of occupational injuries and occupational health problems such as work-related musculoskeletal disorders, mental disorders, and overwork. However, the current research on occupational safety and health of workers in new business forms is insufficient, and the experience of institutional practice and supervision is also limited. Therefore, based on relevant research progress at home and abroad, this paper discussed the health injuries of workers in new business forms, summarized associated influencing factors from four aspects: employment form, employment affiliation, long working hours exposure, and institutional guarantee, and then proposed countermeasures to strengthen occupational safety and health protection of workers. For the current problems in the employment of new business forms, the government, enterprises, and workers need to work together to improve the legal and regulatory system for employment of new business forms, and standardize occupational safety and health management, thus effectively protect the occupational safety and health rights and interests of workers in new business forms.

Helminth infections may trigger host innate and adaptive immune responses. Group 2 innate lymphoid cells (ILC2) are an important factor involved in type 2 immune responses, and produce a large number of T helper 2 cell (Th2) cytokines following stimulation by interleukin (IL)-25, IL-33 and thymic stromal lymphopoietin (TSLP), which play a critical role in parasite clearance and tissue repair. Following helminth infections, autocrine factors, mast cells, enteric nervous system and Th2 cells have been recently found to be involved in regulation of ILC2. Unraveling the role of ILC2 in immune response against helminth infections is of great value for basic research and drug development. This review summarizes the research progress on ILC2 and its role in helminth infections.