[Call for attention to the scientific issues in cardiovascular adverse effects associated with immune checkpoint inhibitors].

Cardiovascular disease (CVD) and cancer are two intimately interconnected conditions with leading causes of mortality worldwide. Emerging evidence suggests that CVD and cancer have common risk factors and share genetics and molecular mechanisms. With recent advancements in diagnosis and treatment, the number of long-term survivors have been continuously increasing. However, cancer patients have significantly higher cardiovascular mortality than general population, mostly resulting from cardiotoxic side effects of anticancer treatments. The prevalence and severity of cardiotoxicity and vascular abnormalities led to the emergence of a clinical branch of cardiology, known as cardio-oncology. Immune checkpoint inhibitors (ICIs) are currently widely used for treatment of various types of cancers. Recent studies suggest that ICIs lead to cardiotoxicity including myocarditis with an incidence of 0.04%-2.4% and a mortality of 25%-50%. However, the molecular and pathophysiologic mechanisms underlying the cardiovascular toxicity induced by ICIs are poorly understood. Therefore, this article combines the recent research results of the pathophysiology of cardiovascular toxicity induced by ICIs and explores novel diagnostic, monitoring, and therapeutic approaches to improve cardiac function and prevent cardiovascular injury.

[Effects of rapid drug sensitivity testing for multidrug-resistant bacteria on the prognosis of patients with severe intra-abdominal infection].

Objective: To examine the clinical value of rapid detection of drug-resistant bacteria by immunochromatography and the effects of rapid detection on the prognosis of patients with severe intra-abdominal infection complicated by carbapenem-resistant Enterobacteriaceae (CRE) bloodstream infection. Methods: This was a retrospective cohort study. We analyzed clinical data of 73 patients with severe abdominal infections with sepsis or septic shock complicated by CRE bloodstream infection admitted to the general surgery department of Jinling Hospital between February 2022 and February 2023. Patients were divided into a colloidal gold immunochromatographic assay (GICA) group (17 patients) and conventional testing group (56 patients) based on whether a GICA for CRE had been performed on the patients' first blood culture sample during the diagnosis and treatment process. There were no statistically significant differences between the GICA and conventional testing groups in age ([55.9±17.3] vs. [47.6±16.4] years), sex ([16 men vs. one woman ] vs. [41 men vs. 15 women]), median Charlson comorbidity index (3.0[2.0,4.0] vs. 3.0[2.0, 4.8]), septic shock (10 vs. 39), or acute kidney injury (8 vs. 40) (all P>0.05). Both groups routinely underwent traditional bacterial identification and drug susceptibility testing. Additionally, patients in the GICA group were tested directly for positive blood cultures using a GICA carbapenemase test kit. The main outcomes were mortality rates on Days 28 and 90 after the first identification of CRE bloodstream infection in both groups. We also compared the microbial clearance rate, duration of hospitalization and intensive care unit stay, and time from onset of CRE bloodstream infection to initiation of targeted and appropriate antibiotics between the two groups. Results: The rate of microbial clearance of bloodstream infection was significantly greater in the GICA group than in the conventional testing group (15/17 vs. 34/56 [60.7%], χ2=4.476, P=0.034), whereas the 28-day mortality tended to be lower in the GICA than conventional testing group [5/17 vs. 44.6% [25/56], χ2=1.250, P=0.264). The 90-day mortality (8/17 vs. 53.6% [30/56], χ2=0.222, P=0.638), median duration of hospitalization (37.0 [18.0, 46.5] days vs. 45.5 [32.2, 64.8] days, Z=-1.867, P=0.062), and median duration of intensive care unit stay (18.0 [6.5, 35.0] days vs. 32.0 [5.0, 51.8] days, Z=-1.251, P=0.209). The median time between the onset of bloodstream infection and administration of antibiotics was 49.0 (38.0, 69.0) hours in the GICA group, which is significantly shorter than the 163.0 (111.8, 190.0) hours in the conventional testing group (Z=-5.731, P<0.001). The median time between the onset of bloodstream infection and administration of appropriate antibiotics was 40.0 (34.0, 80.0) hours in the GICA group, which is shorter than in the conventional testing group (68.0 [38.2, 118.8]) hours; however, this difference is not statistically significant (Z=-1.686, P=0.093). Conclusions: GICA can provide information on carbapenemase- producing pathogens faster than traditional drug sensitivity testing, enabling early administration of the optimal antibiotics. The strategy of 'carbapenemase detection first' for managing bacterial infection has the potential to improve prognosis of patients and reduce mortality rate.

[Treatment of open abdomen combined with entero-atmospheric fistula: A retrospective study].

Objective: The purpose of this study was to analyze the course and outcome of patients with combined entero-atmospheric fistulas in open abdomen treatment. Methods: In this retrospective observational study, we collected data on 214 patients with open abdomen complicated by entero-atmospheric fistulas admitted to Research Institute of General Surgery, Jinling Hospital, Affiliated Hospital of Medical School from January 2012 to January 2021. We collected their basic characteristics, aetiology, treatment plan, and prognosis, including the durations of hospitalization and open treatment, time to resumption of enteral nutrition, duration and prognosis of definitive surgery, and overall prognosis. Results: Of the 214 patients with open abdomen complicated with entero-enteral fistulas, 23 (10.7%) died (11 of multiple organ failure caused by abdominal infection, five of abdominal cavity bleeding, four of pulmonary infection, one of airway bleeding, one of necrotizing fasciitis, and one of traumatic brain injury). The remaining 191 underwent definitive surgery at our hospital. The patients who underwent definitive surgery were predominantly male (156 patients, 81.7%); their age was (46.5±2.5) years. Trauma and gastrointestinal tumors (120 cases, 62.8%) predominated among the primary causes. The reasons for abdominal opening were, in order, severe abdominal infection (137 cases, 71.7%, damage control surgery (29 cases, 15.2%), and abdominal hypertension (25 cases, 13.1%). Temporary abdominal closure measures were used to classify the participants into a skin-only suture group (104 cases) and a skin-implant group (87 cases). Compared with the skin-implant group, in the skin-suture-only group the proportion of male patients was lower (74.7% [65/87] vs. 87.5% [91/104], χ2=5.176, P=0.023), the mean age was older ([48.3±2.0] years vs. [45.0±1.9] years, t=-11.671, P<0.001), there were fewer patients with trauma (32.2% [28 /87] vs. 58.7% [61/104), χ2=13.337, P<0.001), intensive care stays were shorter ([8.9±1.0] days vs. [12.7±1.6] days, t=19.281, P<0.001), total length of stay was shorter ([29.3±2.0] days vs. [31.9±2.0] days, t=9.021,P<0.001), there was a higher percentage of colonic fistulas (18.4% [16/87] vs. 8.7% [9/104], χ2=3.948, P=0.047), but fewer multiple fistulas (11.5% [10/87] vs. 34.6% [36/104], χ2=14.440, P<0.001). As to fistula management, a higher percentage of fistula sealing methods using 3D-printed intestinal stents were implemented in the skin-only suture group (60.9% [53/87] versus 43.3% [45/104], χ2=5.907, P=0.015). Compared with the implant group, the skin-only suture group had a shorter mean time to performing provisional closure ( [9.5±0.8] days vs. [16.0±0.6] days, t=66.023, P<0.001), shorter intervals to definitive surgery ( [165.0±10.7] days vs. [198.9±8.3] days, t=26.644, P<0.001), and less use of biopatches (56.3% [49/87) vs. 71.2% [74/104], χ2=4.545, P=0.033). Conclusions: Open abdomen complicated with entero-enteral fistulas is more common in male, and is often caused by trauma and gastrointestinal tumor. Severe intra-abdominal infection is the major cause of open abdomen, and most fistulae involves the small intestine. Collection and retraction of intestinal fluid and 3D-printed entero-enteral fistula stent sealing followed by implantation and skin-only suturing is an effective means of managing entero-enteral fistulas complicating open abdominal cavity. Earlier closure of the abdominal cavity with skin-only sutures can shorten the time to definitive surgery and reduce the rate of utilization of biopatches.

[Hsp90 participates in the necroptosis of mouse neural cells induced by aluminum through the RIP1/RIP3/MLKL pathway].

Objective: To investigate whether heat shock protein 90 (HSP90) participates in the necroptosis of C57BL/6 mouse neurons and spatial memory impairment induced by Aluminum maltol [Al (mal) (3)] through RIP1/RIP3/MLKL pathway. Methods: In March 2022, Thirty-two C57 mice were randomly divided into control group, Low dose group, a medium dose group, and a high-dose group, with 8 mice in each group, and injected intraperitoneally with physiological saline, 20, 40, and 80, respectivelyµmol/kgAl (mal) (3) was administered, it was injected 5 days a week and discontinued 2 days for 60 days. Morris water maze test was used to test the spatial learning and memory ability of mice. Nissl staining was used to observe the pathological changes of brain tissue. The protein expression levels of RIP1, RIP3, MLKL and HSP90 in hippocampus were determined by Western blotting. Results: In the water maze experiment, compared with the control group, the number of mice crossing the platform decreased in each dose group, the difference was statistically significant (H=9.50, P=0.023), and the number of mice crossing the platform was statistically significant among each dose group (P <0.05). Compared with the control group, the number of hippocampal nerve cells in each dose group decreased, the arrangement was disordered, and the Nissellite bodies decreased. Western blotting results showed that compared with the control group, the expression level of RIP1 protein in the hippocampus of mice in high-dose group was higher, and the difference was statistically significant (P <0.05). The expression levels of RIP3, MLKL and HSP90 in hippocampal tissue of mice in medium and high dose groups were increased, and the differences were statistically significant (P<0.05). After siRNA intervention decreased the expression of HSP90 protein, the expressions of HSP90, RIP1, RIP3 and MLKL in Al (mal) (3) groups were increased, and the differences were statistically significant (P<0.05) . Conclusion: Through RIP1/RIP3/MLKL pathway, HSP90 is involved in neuronal programmed necrosis and spatial memory impairment induced by maltol aluminum in C57 mice.

[A retrospective controlled study on the treatment effect of distraction osteogenesis and maxillomandibular advancement for severe obstructive sleep apnea hypopnea syndrome patients].

Objective: To compare the treatment effect of distraction osteogenesis (DO) and maxillomandibular advancement (MMA) for severe obstructive sleep apnea hypopnea syndrome (OSAHS) patients and to guide clinical decisions about treatment of OSAHS. Methods: Thirty-seven OSAHS patients which accepted maxillomandibular advancement (MMA) or distraction osteogenesis (DO) in Stomatological Hospital of the Department of Maxillofacial Trauma and Orthognathic Surgery, School of Stomatology, The Forth Military Medical University from June 2017 to June 2019 were collected. Their preoperative and postoperative data of cephalometry, polysomnography (PSG), Pittsburgh sleep quality index (PSQI) and Epworth sleepiness scale (ESS) scores were collected and analyzed. With propensity score matching method, the treatment effect of MMA and DO was analyzed and compared. Results: According to the statistics of MMA group, only AHI was correlated with operative successful rate and cure rate. With the increase of AHI, the treatment effect of MMA on OSAHS patients gradually decreased. The cut-off point of AHI as a predictor of MMA treatment failure was 78.2 n/h. All the matched cases were severe OSAHS patients. Statistical analysis showed that the mandibular elongation of DO patients[(24.00±4.39) mm] was significantly more than that of MMA group [(11.20±1.37) mm] (t=-6.11, P<0.001), the improvement of PSG index [including lowest oxygen saturation (LSpO2), longest apnea (LA) and longest hypopnea (LH)] in DO group [LSpO2=(93.40±1.82)%; LA=(18.28±8.32) s; LH=(61.84±32.94) s] was significantly higher than that in the MMA group [LSpO2=(86.00±4.06)%, LA=(64.08±21.78) s, LH=(172.40±30.70) s](t=-3.72, P=0.005; t=4.39, P=0.003; t=5.49, P=0.004). The PSQI and the ESS scores of DO group (PSQI=4.20±0.83; ESS=3.40±1.52) were also significantly better than that of MMA group (PSQI=8.80±2.39, ESS=9.40±2.88)(t=4.07, P=0.001; t=4.12, P=0.002). Conclusions: For severe OSAHS patients, the objective and subjective indicators of DO treatment group showed a better therapeutic effect than that of MMA.

[The feasibility study of objective evaluation of the severity of motion sickness by quantitative analysis of the facial skin color].

Objective: To explore the feasibility of applying quantitative analysis of the facial skin color to evaluate the severity of motion sickness objectively and to seek objective indicators that can reflect the severity of motion sickness. Methods: Motion sickness was induced in 51 male adult subjects recruited at the Air Force Medical University by Coriolis acceleration stimulation, and facial skin colorimetric values were acquired using a portable spectrophotometer at five time points: before stimulation and at 0 min, 10 min, 20 min and 30 min after the end of stimulation. The Graybiel rating scales were applied to assess the severity of motion sickness in subjects at each time point after stimulation, and the correlation between the magnitude of change in each colorimetric value and the maximum Graybiel's score was analyzed. The ROC curves were used to compare the evaluation performance of colorimetric value indicators which could reflect the severity of motion sickness. Results: Each colorimetric value in the CIE-L*a*b* color system changed significantly after exposure to provocative motion stimuli, and the trend was consistent with the typical sign of pallor in motion sickness. The magnitudes of the increase in the colorimetric value CIE-L*, the decrease in CIE-a*, and the increase in CIE-b* were all significantly and positively correlated with the maximum of Graybiel's scores (r=0.490 0, P=0.000 3; r=0.549 3, P<0.000 1; r=0.540 9, P<0.000 1). Comparing the performance of three colorimetric indicators to assess the severity of motion sickness, CIE-a* had an area under the ROC curve of 0.875 0, a sensitivity of 85.71%, and a specificity of 87.50%, which was better than CIE-L* and CIE-b*. Conclusions: The CIE-L*a*b* colorimeter values can be considered as objective indicators of the severity of motion sickness, among which the colorimetric indicator CIE-a* has the most diagnostic significance, and the method of quantitative analysis of the facial skin color can provide a new reference for the objective evaluation of the severity of motion sickness.

[Regulatory mechanism of deferoxamine on macrophage polarization and wound healing in mice with deep tissue injury].

Objective: To investigate the effects of deferoxamine on macrophage polarization and wound healing in mice with deep tissue injury (DTI) and its mechanism. Methods: The experimental research methods were adopted. Fifty-four male C57BL/6J mice of 6-8 weeks old were divided into DTI control group, 2 mg/mL deferoxamine group, and 20 mg/mL deferoxamine group according to random number table, with 18 mice in each group. DTI was established on the back of mice by magnet compression method. From post injury day (PID) 1, mice were injected subcutaneously with 100 µL normal saline or the corresponding mass concentration of deferoxamine solution every other day at the wound edge until the samples were collected. Another 6 mice without any treatment were selected as normal control group. Six mice in each of the three DTI groups were collected on PID 3, 7, and 14 to observe the wound changes and calculate the wound healing rate. Normal skin tissue of mice in normal control group was collected on PID 3 in other groups (the same below) and wound tissue of mice in the other three groups on PID 7 and 14 was collected for hematoxylin-eosin (HE) staining to observe the tissue morphology. Normal skin tissue of mice in normal control group and wound tissue of mice in the other three groups on PID 7 were collected, and the percentages of CD206 and CD11c positive area were observed and measured by immunohistochemical staining, and the mRNA and protein expressions of CD206, CD11c, and inducible nitric oxide synthase (iNOS) were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction and Western blotting, respectively. Normal skin tissue of mice in normal control group and wound tissue of mice in DTI control group and 20 mg/mL deferoxamine group were collected on PID 3, 7, and 14, and the protein expressions of signal transducer and activator of transcription 3 (STAT3) and interleukin-10 (IL-10) were detected by Western blotting. The sample number in each group at each time point in the above experiments. The RAW264.7 cells were divided into 50 µmol/L deferoxamine group, 100 µmol/L deferoxamine group, 200 µmol/L deferoxamine group, and blank control group, which were treated correspondingly, with 3 wells in each group. The positive cell percentages of CD206 and CD86 after 48 h of culture were detected by flow cytometry. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and least significant difference test. Results: On PID 7, the wound healing rates of mice in 2 mg/mL and 20 mg/mL deferoamine groups were (17.7±3.7)% and (21.5±5.0)%, respectively, which were significantly higher than (5.1±2.3)% in DTI control group (P<0.01). On PID 14, the wound healing rates of mice in 2 mg/mL and 20 mg/mL deferoamine groups were (51.1±3.8)% and (57.4±4.4)%, respectively, which were significantly higher than (25.2±3.8)% in DTI control group (P<0.01). HE staining showed that the normal skin tissue layer of mice in normal control group was clear, the epidermis thickness was uniform, and skin appendages such as hair follicles and sweat glands were visible in the dermis. On PID 7, inflammation in wound tissue was obvious, the epidermis was incomplete, and blood vessels and skin appendages were rare in mice in DTI control group; inflammatory cells in wound tissue were reduced in mice in 2 mg/mL and 20 mg/mL deferoxamine groups, and a few of blood vessels and skin appendages could be seen. On PID 14, inflammation was significantly alleviated and blood vessels and skin appendages were increased in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups compared with those in DTI control group. On PID 7, the percentages of CD206 positive area in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly higher than that in DTI control group (P<0.01), the percentage of CD206 positive area in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue of mice in normal control group (P<0.01), the percentage of CD206 positive area in wound tissue of mice in 20 mg/mL deferoxamine group was significantly higher than that in normal skin tissue of mice in normal control group (P<0.01). The percentages of CD11c positive area in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly lower than those in DTI control group and normal skin tissue in normal control group (P<0.05 or P<0.01), and the percentage of CD11c positive area in normal skin tissue of mice in normal control group was significantly higher than that in DTI control group (P<0.05). On PID 7, the CD206 mRNA expressions in the wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly higher than that in DTI control group (P<0.01), but significantly lower than that in normal skin tissue in normal control group (P<0.01); the CD206 mRNA expression in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue in normal control group (P<0.01). The mRNA expressions of CD11c and iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly lower than those in DTI control group (P<0.01). The mRNA expressions of CD11c in the wound tissue of mice in DTI control group, 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than that in normal skin tissue in normal control group (P<0.01). Compared with that in normal skin tissue in normal control group, the mRNA expressions of iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly decreased (P<0.01), and the mRNA expression of iNOS in wound tissue of mice in DTI control group was significantly increased (P<0.01). On PID 7, the protein expressions of CD206 in the wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than those in DTI control group and normal skin tissue in normal control group (P<0.01), and the protein expression of CD206 in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue in normal control group (P<0.01). The protein expressions of CD11c and iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly lower than those in DTI control group (P<0.01). The protein expressions of CD11c and iNOS in wound tissue of mice in DTI control group were significantly higher than those in normal skin tissue in normal control group (P<0.01). The CD11c protein expressions in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than those in normal skin tissue in normal control group (P<0.05 or P<0.01). The protein expression of iNOS in wound tissue of mice in 2 mg/mL deferoamine group was significantly lower than that in 20 mg/mL deferoamine group and normal skin tissue in normal control group (P<0.05). On PID 3, 7, and 14, the protein expressions of STAT3 and IL-10 in wound tissue of mice in 20 mg/mL deferoxamine group were significantly higher than those in DTI control group (P<0.05 or P<0.01), and the protein expressions of STAT3 were significantly higher than those in normal skin tissue in normal control group (P<0.05 or P<0.01). On PID 7 and 14, the protein expressions of IL-10 in wound tissue of mice in 20 mg/mL deferoxamine group were significantly higher than those in normal skin tissue in normal control group (P<0.01). On PID 3, 7, and 14, the protein expressions of IL-10 in wound tissue of mice in DTI control group were significantly lower than those in normal skin tissue in normal control group (P<0.05 or P<0.01). After 48 h of culture, compared with those in blank control group, the CD206 positive cell percentages in 100 µmol/L and 200 µmol/L deferoamine groups were significantly increased (P<0.01), while the CD86 positive cell percentages in 100 µmol/L and 200 µmol/L deferoamine groups were significantly decreased (P<0.01). Conclusions: Deferoxamine can promote the polarization of macrophages toward the anti-inflammatory M2 phenotype and improve wound healing by enhancing the STAT3/IL-10 signaling pathway in DTI mice.

Tazarotene gel promotes healing of deep tissue injury in mice.

We investigated the efficacy and molecular mechanisms of tazarotene gel for healing deep tissue injury (DTI). We used male C57BL/6J mice to establish a DTI model. Animals were divided randomly into control, tazarotene gel and purilon gel groups. We injected 100 ul tazarotene gel, purilon gel or saline every 48 h for 20 days. Hematoxylin and eosin staining was used to observe pathological changes on days 14 and 21. The mRNA and protein expression of VEGF-α, TGF-ß1 and HIF-1α were detected by qRT-PCR and western blot, respectively. Wound sites exhibited accelerated healing by 20 days in the tazarotene gel group. Fewer inflammatory cells and more granulation tissue were found in both experimental groups compared to controls. The mRNA and protein expression of VEGF-α and TGF-ß1 in the experimental groups were increased compared to the control group by day 14. Expression of HIF-1α in the experimental groups was significantly less than in the controls. Tazarotene gel promoted wound healing independent of the HIF-1α/VEGF signalling pathway during tissue repair of DTI. Tazarotene and purilon gels exhibited similar macroscopic healing of wounds and expression of genes and proteins.

[Expression of miR-495 and its effect on MHCC-97H hepatocellular carcinoma cells].

Objective: To investigate the expression of miR-495 and its effect on MHCC-97H hepatocellular carcinoma cells. Methods: Fifty-six hepatocellular carcinoma tissue specimens (HCC group) and 40 normal liver tissue specimens (control group) preserved in our hospital from January 2017 to January 2018 were selected. Reverse transcription real-time fluorescent quantitative PCR (qRT-PCR) was used for miR-495 expression detection. MHCC-97H HCC cells were randomly selected and then divide into control group, blank plasmid group and transfection group. The blank plasmid group was transfected with blank plasmid, and the transfection group was transfected with miR-495 inhibitor. The expression of miR-495 in each group of cells were detected using qRT-PCR. CCK method was used to detect each group proliferation activity. Transwell cell migration assay was used to detect each group migration ability. Analysis of variance was used for comparison between multiple groups. Furthermore, LDS-t test was used for pairwise comparison, and t -test was used for comparison between the two groups. Results: The relative expression levels of miR-495 in the HCC group was (2.043 ± 0.382), which was higher than the control group, and the difference between the two groups was statistically significant (P < 0.05). The relative expressions levels of miR-495 in patients with stage III to IV and lymph node metastasis were 2.265 ± 0.284 and 2.290 ± 0.355, which were significantly higher than those of stage I to II and no lymph node metastasis (P < 0.05). The relative expression levels of miR-495 in transfection group was 0.653 ± 0.102, which were significantly lower than control group and blank plasmid group (P < 0.05). The A values of MHCC-97H cells cultured for 24 h and 48 h in transfection group were 0.404 ± 0.106 and 0.604 ± 0.136, which were significantly lower than control group and blank plasmid group (P < 0.05). MHCC-97H cells migration number in the transfection group was (6.10 0 ± 20), which was significantly lower than that of control group and blank plasmid group (P < 0.05). Conclusion: miR-495 high expression has certain relationship with clinicopathological characteristics of HCC tissues. In addition, miR-495 has a certain effect on the proliferation and migration ability of MHCC-97H HCC cells.

[Quantitative study of T1 weighted imaging labyrinthine high signal ratio based on magnetic resonance imaging in patients with unilateral sudden deafness accompanied by vertigo and tinnitus].

Objective: To explore the value of speculating etiology of the magnetic resonance imaging (MRI) T1 weighted imaging (T1WI) labyrinthine high signal ratio in patients with unilateral sudden deafness accompanied by vertigo and tinnitus and its relationship with hearing prognosis. Methods: Fifty-two patients with unilateral sudden deafness accompanied by vertigo and tinnitus who were admitted to Beijing Tongren Hospital Affiliated to Capital Medical University from January 2016 to July 2019 were collected, including 27 males and 25 females, aged (47.7±15.1) years. The inner ear MRI data of 52 patients (17 plain scan, 35 enhanced scan) with unilateral sudden deafness were retrospectively analyzed. Two radiologists independently measured the labyrinthine high signal intensity of the affected side and the contralateral normal side on T1WI and enhanced T1WI and calculated the signal ratio (the normal labyrinth signal was subtracted from the affected signal and then divided by the normal signal). The etiology of the enhanced group was judged based on two methods, including whether the abnormal high signal was enhanced or not (unenhancement indicated hemorrhage and enhancement indicated inflammation), and the locations of labyrinthine involvement on enhanced three-dimensional fluid attenuated inversion recovery (3D-FLAIR) (inflammation usually involved the perilymph spaces, while hemorrhage involved the perilymph and endolymph spaces). In the plain group, the locations of labyrinthine involvement on 3D-FLAIR was applied to infer the potential etiology. Results: The two methods presumed that 8 cases might be hemorrhage (22.9%, 8/35) and 27 be inflammation (77.1%, 27/35) in the enhanced group, which had a high consistency, while it was speculated that 7 patients might be hemorrhage (7/17) and 10 patients be inflammation (10/17) in the plain group. The measurement results of the two radiologists were highly consistent within and between the groups [the intraclass correlation coefficient (ICC) values were greater than 0.800]. The area under the receiver operating characteristic (ROC) curve (AUC) of the T1WI high signal ratio in the enhanced group for speculating etiology was 0.949 (P<0.01), when the predictive threshold value was 0.467, with a sensitivity of 96.3% and a specificity of 87.5%. It might be hemorrhage when the ratio was higher than the threshold value, otherwise it was inflammation. The T1WI labyrinthine high signal ratio was higher in the hemorrhage group than that of the inflammation group, and the hearing prognosis was worse (all P<0.05). The T1WI labyrinthine high signal ratio of the unrecovered group was higher than that of the recovered group (P=0.034). Conclusions: The etiology of labyrinthine high signal formation can be inferred by quantitative values combined with the involved sites. The high signal in the labyrinth indicates poor hearing prognosis, the higher the signal intensity, the greater the possibility of hemorrhage and the worse the hearing prognosis.

Identification of the most appropriate existing anthropometric index for home-based obesity screening in children and adolescents.

OBJECTIVES: Home-based self-screening and monitoring for obesity is particularly valuable for the prevention and control of chronic diseases. This study aimed to identify an anthropometric index suitable for home-based obesity screening in children and adolescents. STUDY DESIGN: The design of this study is a cross-sectional study. METHODS: A total of 14,042 students (6-17 years) from the Qibao Community, Minhang District, Shanghai, were studied in 2018. The percentage body fat (PBF), height, weight, waist circumference (WC) and hip circumference were measured. Body mass index (BMI), triponderal mass index (TMI), body adiposity index (BAI), waist-to-hip ratio (WHR) and waist-to-height ratio (WHtR) were calculated. Partial correlation analysis was used to evaluate the relationships between these indices and PBF, and receiver operating characteristic (ROC) curves were used to evaluate their performance for obesity screening. RESULTS: BMI, TMI, WC and WHtR were found to strongly correlate with PBF (r ≥ 0.830, all P < 0.001). The optimal index for obesity screening in children (6-11 years) was BMI (area under the ROC curve [AUC] = 0.980 for boys and 0.981 for girls) and in adolescents (12-17 years) was TIM (AUC = 0.976 for boys and 0.945 for girls); however, the optimal cut-off values for BMI and TMI differed among the subgroups. The ROC curve analysis showed that WHtR had similar cut-off values in each subgroup (0.45 for boys of 6-11 years and 0.43 for the other subgroups), excellent performance in children (AUC>0.90) and good performance in adolescents (AUC = 0.960 for girls and 0.878 for boys). CONCLUSIONS: Owing to its accuracy and stable cut-off value for defining obesity, WHtR should be recommended for home-based obesity screening in children and adolescents.

MiR-33a-5p inhibits the growth and metastasis of melanoma cells by targeting SNAI2.

MicroRNAs have been verified as critical regulators in the development of melanoma. miR-33a-5p was significantly downregulated in melanoma, however, the specific role and regulatory mechanism of miR-33a-5p in melanoma were still unclear. The present study identified that miR-33a-5p was downregulated in melanoma tissues and cells, while SNAI2 was upregulated. miR-33a-5p directly targeted SNAI2 and negatively regulated its expression in melanoma cells. Overexpression of miR-33a-5p repressed proliferation, migration, invasion, EMT and promoted apoptosis of melanoma cell in vitro, these effects were partially reversed by SNAI2 overexpression. In addition, miR-33a-5p impaired melanoma growth in vivo by inhibiting SNAI2. Mechanistically, miR-33a-5p repressed activation of the PI3K/AKT/mTOR pathway by targeting SNAI2. In conclusion, miR-33a-5p repressed the progression of melanoma by targeting SNAI2 via inactivation of the PI3K/AKT/mTOR signaling pathway, providing a potential molecular mechanism for the treatment of melanoma.

[Subcellular localization of GTPase of immunity-associated protein 2].

OBJECTIVE: To analyze the subcellular localization of GTPase of immunity-associated protein 2 (GIMAP2) for the further functional study. METHODS: In the study, we first obtained the protein sequences of GTPase of immunity-associated protein 2 (GIMAP2) from National Center for Biotechnology Information (NCBI) database, and then performed a prediction analysis of its transmembrane structure, nuclear localization signal (NLS), nuclear export signal (NES) and subcellular localization through bioinformatics online tools. GIMAP2 gene amplified by PCR was inserted into the expression vector pQCXIP-mCherry-N1 and positive clones were selected by ampicillin resistance. After using methods to extract and purify, the sequenced recombinant plasmid pQCXIP-GIMAP2-mCherry, together with the retroviral packaging plasmids VSVG and Gag/pol, was transferred into HEK293FT cells by liposomes for virus packaging. The virus supernatant was collected 48 h after transfection and directly infected the human breast cancer cell line MDA-MB-436. Immunofluorescence staining was constructed to detect the localization of endogenous and exogenous GIMAP2 in MDA-MB-436 cells. Meanwhile, green fluorescent chemical dyes were used to label mitochondria, endoplasmic reticulum, and lipid droplets in living MDA-MB-436 cells stably expressing the GIMAP2-mCherry fusion protein. Images for the three dye-labeled organelles and GIMAP2-mCherry fusion protein were captured by super-resolution microscope N-SIM. RESULTS: Bioinformatics analysis data showed that GIMAP2 protein composed of 337 amino acids might contain two transmembrane helix (TM) structures at the carboxyl terminus, of which TMs were estimated to contain 40-41 expected amino acids, followed by the residual protein structures toward the cytoplasmic side. NES was located at the 279-281 amino acids of the carboxyl terminus whereas NLS was not found. GIMAP2 might locate in the lumen of the endoplasmic reticulum. Sequencing results indicated that the expression vector pQCXIP-GIMAP2-mCherry was successfully constructed. Fluorescent staining confirmed that GIMAP2-mCherry fusion protein, co-localized well with endogenous GIMAP2, expressed successfully in the endoplasmic reticulum and on the surface of lipid droplets in MDA-MB-436 cells. CONCLUSION: GIMAP2 localizes in the endoplasmic reticulum and on the surface of LDs, suggesting potential involvement of GIMAP2 in lipid metabolism.

[A clinical performance study of modified CT angiography in detecting bronchial artery-pulmonary artery fistula].

Objective: To evaluate the clinical value of modified computed tomography angiography(CTA) in detecting bronchial artery-pulmonary artery fistula(BPF). Methods: Retrospective analysis was performed on 246 patients with hemoptysis admitted to the First Affiliated Hospital of Wenzhou Medical University from July 2017 to December 2018, who underwent modified CTA and DSA examination at the same time. CT was performed with Toshiba Aquilion one 320 row 640-slice spiral CT scanner. All modified CTA images were read blindly by two radiologists above the attending doctors. The sensitivity, specificity and accuracy of the modified CTA in diagnosing BPF were calculated with the DSA results as the reference,and the consistency of the two tests was analyzed. Results: DSA detected 186 cases of positive and 60 cases of negative, modified CTA detected 160 cases of positive and 86 cases of negative. The sensitivity,specificity and accuracy of modified CTA for BPF diagnosis was 85.5%(159/186),98.3%(59/60), 88.6%(218/246) respectively, and they were with high consistency with DSA examination results (kappa=0.73,P<0.01). Conclusion: Modified CTA has high diagnostic specificity for BPF,which can be used as the preferred method for non-invasive screening of suspected BPF patients.

Application of 18S rDNA Clone Library to Detect Diatom Population Diversity in Dianchi.

ABSTRACT: Objective To detect the diatom population diversity in Dianchi by constructing a 18S rDNA clone library. Methods DNA from diatoms in 6 water samples of Dianchi was amplified with diatom 18S rDNA specific primer.The 18S rDNA clone library was constructed, and clones were randomly selected for sequence. Sequence alignment was performed by BLAST. The diatom population distribution in Dianchi was analyzed and the phylogenetic tree of diatom 18S rDNA in Dianchi waters was established with the MEGA v7.0.14 software. Results Two hundred and forty clones were sequenced, with 167 diatom sequences obtained, including 11 diatom species such as Stephanodiscus, Diatoma, and Melosira. There were certain differences in diatom population distribution among the 6 samples. Conclusion The population distribution of diatom species in Dianchi shows unique features and the sequence analysis of diatom 18S rDNA has a certain reference value to the inference of forensic drowning sites.

Clinicopathological and prognostic significance of miR-4317 expression in gastric cancer patients.

OBJECTIVE: Previous studies have revealed that miR-4317 was abnormally expressed and functioned as a tumor suppressor in several tumors, including gastric cancer (GC). However, the clinical significance of miR-4317 in GC remains largely unclear. Our present study aimed at investigating the possibility of miR-4317 as a novel prognostic biomarker for GC patients. PATIENTS AND METHODS: RT-PCR was performed to determine the levels of miR-4317 in GC tissues and matched normal gastric tissues. Association between miR-4317 levels and clinicopathological factors was also analyzed using Chi-square test. Kaplan-Meier survival analysis and univariate and multivariate assays were further conducted to investigate the correlation between miR-4317 expression and GC patients prognosis. RESULTS: We showed that miR-4317 expression levels were significantly lower in GC cell lines compared to matched normal tissues (p< 0.01). Down-regulation of miR-4317 was observed to be associated with lymph node metastasis (p = 0.019), distant metastasis (p = 0.011) and TNM stage (p = 0.007). In addition, Kaplan-Meier analysis showed that patients with low miR-4317 expression had a shorter overall survival compared with the high miR-4317 expression group (p = 0.0009). Furthermore, multivariate analysis revealed that miR-4317 expression was an independent prognostic marker of overall survival of GC patients. CONCLUSIONS: miR-4317 expression may be a novel and valuable prognostic factor in GC.

[Magnetic resonance imaging study of endolymphatic hydrops based on 3D-FLAIR sequence after single-dose intravenous gadolinium injection].

Objective: To investigate the visualization of endolymph in patients with otogenic vertigo by intravenous administration of single dose of gadolinium contrast agents and magnetic resonance three-dimensional fluid-attenuated inversion recovery sequence (3D-FLAIR MRI), and further assess the extent of endolymphatic hydrops. Methods: From Beijing Tongren Hospital of Capital Medical University between October 2017 and June 2018, 30 patients (16 males, 14 females) with unilateral otogenic vertigo were involved in this study, with the age of 30 to 68 years, mean age of (53±10) years. Eight hours after intravenous administration of single dose (0.1 mmol/kg, body weight) of gadopentetate (Gd-DTPA), 3D-FLAIR sequence was performed in 30 patients. The location of endolymphatic hydrops was observed and then the degree of hydrops was quantitatively elevated by two radiologists. The consistency test was used to analyze the location and degree of endolymphatic hydrops in the two radiologists and the paired t-test was used to compare the difference between the affected and healthy side of endolymphatic spaces of the patients with otogenic vertigo. Results: In 30 patients, the gadolinium distributed in all parts of the perilymph inside the inner ear, and can accurately outline the boundaries of the peri-and endolymph. Twenty-six patients (26/30, 86.7%) were found to have unilateral endolymphatic hydrops, including 18 mild hydrops, 8 significant hydrops. The two radiologists had a very good agreement on the assessment of endolymphatic hydrops(kappa=0.864, ICC=0.959). In the 3D-FLAIR MR images of 26 patients with endolymphatic hydrops, the saccule (26/26, 100%) had a higher rate of hydrops than the cochlea and utricle(16/26, 61.5%; 14/26, 53.8%), and two radiologists had a very good agreement on the location of endolymphatic hydrops(kappa=0.820). Moreover, there was a significantly statistical difference between the affected and healthy area of the endolymphatic space in this study (P<0.01). Conclusion: The technique of 3D-FLAIR MR imaging through single dose intravenous gadolinium injection is feasible, which can estimate endolymphatic hydrops in patients with otogenic vertigo, and accurately classify the degree of hydrops.

[Construction and improvement of animal models with different positional osseous metastasis of prostate cancer in vivo].

OBJECTIVE: To provide an important tool for the study of diagnose and treatment of prostate cancer (PCa) osseous metastasis and change of bone stress force on prostate cancer (PCa) osseous metastasis and a platform, which is more congruous to clinical process, for prevention and cure of neoplastic bone metastases, and to carry out the construction and improvement of animal models of PCa with different positional osseous metastasis in vivo. METHODS: Different gradient concentrations of RM-1 cells were inoculated into the cavity of left femoral bone or lumbar vertebra of mice (C57BL/6) respectively. The change of mouse activity, tumor formation, tumor size and survival time were observed respectively. And the femur tissue and spinal tissue were obtained from the mice after death. The gray value of iconography were measured by imageological examination of femur tissue, and the final histopathological examination were taken to determine the tumor type in both femur and spinal tissue. RESULTS: The tumor growth could be touched at the puncture site in all the mice after inoculated for 7 days. There were no obvious differences in the time of tumorigenesis, the rate of tumor growth and tumor size among the mice in the same group (P>0.05). As the result, the construction femoral bone and lumbar vertebra metastatic models of PCa had been confirmed by iconography and pathology detection. At the same time, the survival time of the mice inoculated with low concentrations of PCa cells was obviously longer than that of high concentrations of PCa cells ( at least 2 weeks longer). CONCLUSION: The animal models with different positional osseous metastasis (limbs and axial skeleton) of PCa using the same PCa cells (RM-1) had been first constructed successfully in our study. At the same time, a high success rate of construction of PCa animal model with bone metastasis was obtained by femoral bone marrow cavity injection of PCa cells. The rate of tumor growth was rapid, animal survival time was appropriate, and the PCa animal model with bone metastasis can be stably reproduced by our method. These animal models can be used to explore the pathogenesis of different positional PCa bone metastasis and provide a new platform, which were more congruous to clinical process, for prevention and cure of neoplastic bone metastases.

[Association between variations in protocadherin 15 gene and occupational noise-induced hearing loss].

Objective: The aim of this study was to investigate whether genetic variability in the protocadherin 15 (PCDH15) gene may correspond with increased susceptibility to noise-induced hearing loss (NIHL) in a Chinese population. Methods: A nested case-control study was performed that followed a cohort of 7 445 noise-exposed workers in a steel factory of Henan province in China from January 1, 2006 to December 31, 2015. In this study, 394 cases who had an average hearing threshold of more than 40 dB (A) in high frequency were defined as the case group, and 721 controls who had an average hearing threshold of less than 35 dB (A) in high frequency and less than 25 dB (A) in speech frequency were defined as the control group. A questionnaire was completed by participants and a physical test was also conducted. SNP genotyping was performed using the SNPscanTM Kit. Multivariate unconditional logistic regression additive models were used to analyze the genotypes in different groups, and the association with NIHL. Unconditional logistic regression models were used to assess the associations between the genotypes and NIHL. Results: The average age of study participants was (40.5±8.3) years and the median number of noise-exposed working years M (P25, P75) was 21.1 (9.1, 27.3). The range of noise exposed levels and the levels of cumulative noise exposure (CNE) were 80.1- 98.8 dB(A) and 86.6- 111.2 dB(A), respectively. Only the distribution of the genotypes (TT/CC/CT) of rs11004085 in the PCDH15 gene showed a significant difference between the case and control groups (P=0.049). In the case group, the distribution was 370 (93.9%), 24 (6.1%) and 0; in the control group, the distribution was 694 (96.3%), 23 (3.2%) and 1 (0.1% ). After smoking, drinking, hypertension, height and CNE adjustment, compared with the TT genotype individuals with the CC/CT genotype had a 1.90-fold increased risk of NIHL (95% CI: 1.06- 3.40). After stratified these data by the noise exposure level or CNE when the noise exposure level was>85 dB (A), compared with cases with the AA genotype of rs10825113, individuals with the GA/GG genotype had a 2.63-fold increased risk of NIHL (95% CI: 1.12- 6.14). When the CNE was ≤ 98 dB(A), compared with cases with the TT genotype of rs11004085, individuals with the CC/CT genotype had a 2.96-fold increased risk of NIHL (95% CI: 1.33- 6.56). However, these differences were not significant after Bonferroni correction had been applied. Conclusions: The results confirmed that genetic variation within the PCDH15 gene may affect the susceptibility to NIHL.

[Association between eye absent homolog 4 gene polymorphisms and occupational noise-induced hearing loss].

Objective: To identify the association between genetic polymorphisms in the eye absent homolog 4 (EYA4) gene and noise-induced hearing loss (NIHL). Method: A nested case control study was conducted based on a cohort of noise-exposed subjects. In total, 292 cases were selected from a steel factory from 6 297 subjects during Jan 1, 2006 to Dec 12, 2015,who had an average hearing threshold of more than 40 dB(A); 584 matched control subjects for each case were designated on the basis of matched criteria including same gender, age (±5 years) and duration of exposure to noise (±2 years). What's more, the control group had an average hearing threshold of less than 35 dB(A) in high frequency and less than 25 dB(A) in speech frequency. Four single nucleotide polymorphisms (SNPs) of the EYA4 gene were genotyped using a SNPscanTM multiplex SNP genotyping kit. Hardy-Weinberg equilibrium tests were performed using a χ2 test for goodness-of-fit for each SNP among the control group, and the effects of genotypes of the EYA4 gene on NIHL were analyzed by logistic regression. The haplotypes were established and their frequencies in the two groups were assessed using Haploview 4.2 and Phase 2.1 software, and interactive effects between haplotypes and cumulative noise exposure were analyzed. Results: The average age of the subjects was (40.1±8.4) years and the average number of noise-exposed working years was 20.3 (8.4, 27.3) years. The range of noise exposure levels and the cumulative noise exposure were 80.2- 98.8 dB (A) and 86.6- 111.2 dB(A) · year, respectively. After adjustment for covariates including height, blood pressure, drinking status and smoking status, in the noise intensity>85 dB (A) group, subjects carrying the rs3813346 TT genotype had a higher NIHL risk than those carrying the GG genotype, and the adjusted OR (95% CI) value was 2.12 (1.21- 3.69). In the cumulative noise exposure>98 dB (A) · year group, compared with haplotype TGC, haplotype CGT showed a protective effect in the development of NIHL, with an adjusted OR (95% CI) value of 0.60 (0.37-0.97), however, the significance of intercation between EY4 gene of noise was lost after Bonferroni correction. Conclusion: Genetic polymorphism in the EYA4 gene may be a genetic susceptibility factor for NIHL.