[ ABSTRACT] AIM:To investigate the relationship between morphological changes of autophagy and apoptosis in the PC12 cells induced by oxygen-glucose deprivation and reoxygenation .METHODS:The PC12 cells were randomly di-vided into normal control group , oxygen-glucose deprivation and reoxygenation group , autophagy inhibitor group and auto-phagy activator group .The cells in oxygen-glucose deprivation and reoxygenation group , autophagy inhibitor group and au-tophagy activator group were exposed to reoxygenation (12 h) after 3 h of oxygen-glucose deprivation, and autophagy inhib-itor 3-methyladenine and autophagy activator rapamycin were added into the cells at the same time .Using transmission elec-tron microscope and monodansylcadaverine fluorescence staining , the morphological changes of autophagosome were ob-served.The apoptosis of the PC12 cells were analyzed by flow cytometry with Annexin V-FITC/PI staining and TUNEL method.RESULTS: Compared with normal control group , the numbers of autophagosomes and the apoptotic rates in-creased in oxygen-glucose deprivation and reoxygenation group (P<0.05).Compared with oxygen-glucose deprivation and reoxygenation group , the numbers of autophagosomes decreased obviously ( P<0.05 ) and the apoptotic rates increased markedly in autophagy inhibitor group (P<0.05).The numbers of autophagosomes increased obviously (P<0.05), the apoptotic rates decreased markedly ( P<0.05 ) , the autophagosomes became bigger in size , and autolysosomes was also found in autophagy activator group .CONCLUSION: Oxygen-glucose deprivation and reoxygenation induce autophagy in PC12 cells, and autophagy inhibits cell apoptosis to play a protective role .

Aim To investigate the effects of astragalo-side IV on apoptosis of PC12 cells inducedby hypoxia/hypoglycemia and reoxygenation. Metheds PC12 cells were randomly divided into 4 groups: normal control group,hypoxia/hypoglycemia and reoxygenation group, astragaloside Ⅳ group and vehicle group. Hypoxia/hy-poglycemia and reoxygenation group, astragaloside Ⅳgroup and vehiclegroup were exposed to reoxygenation (12 h) after 3 h of oxygen and glucose deprivation, and astragaloside Ⅳ was added into cells at the same time. Inverted microscope was used to observe the morphological changes ofPC12 cells and MTT method to detect the activities of PC12 cells, and Annexin V-FITC/PI assay and TUNEL staining were used to meas-ure the apoptosis of PC12 cells. Results Compared with normal control group, cells became round or swol-len and its cellula processes were retracted or disap-peared in hypoxia/hypoglycemia and reoxygenation group;a large number of apoptotic cells could also be observed,whose nucleus were shrinkaged, fragmented or deep-stained. The activities of hypoxia/hypoglycemia and reoxygenation group were decreased markedly than those in normalcontrol group(P<0. 05),while the ap-optotic rates of hypoxia/hypoglycemia and reoxygen-ation group were increased obviously than those in nor-malcontrol group( P<0. 05 ) . Compared with hypoxia/hypoglycemia and reoxygenation group, a good cell growth state could be observed and cellula processes could also be observed significantly in astragaloside Ⅳgroup. The activities of astragaloside Ⅳ group were in-creased than those in hypoxia/hypoglycemia and reoxy-genation group(P<0. 05),while the apoptotic rates of astragalosideⅣgroup were decreased than those in hy-poxia/hypoglycemia and reoxygenation group ( P <0. 05 ) . There was no obvious difference between vehi-clegroup and hypoxia/hypoglycemia and reoxygenation group( P >0. 05 ) . Conclusion Astragaloside IV can reduce the damage of PC12 cells induced by hypoxia/hypoglycemia and reoxygenation, increase cell activity and inhibit cell apoptosis.

OBJECTIVETo explore the effects of hyperbaric oxygen preconditioning (HBOP) on human stress responses during acute exposure to high altitude and the possible mechanism.

METHODSEight male subjects were treated with HBOP for 3, 5, and 7 days, followed by acute exposure to hypoxia simulating an altitude of 4,000 m. Subjects at rest were divided into sea-level control group, simulated high-altitude group, and 5-day HBOP intervention group, while subjects after physical load were divided into sea-level control group, simulated high-altitude group, 3-day HBOP intervention group, and 7-day HBOP intervention group. The physical load test was performed for each subject before and after HBOP, and the plasma levels of dopamine (DA), epinephrine (E), norepinephrine (NE), and adrenocorticotropic hormone (ACTH) were determined before and after exercise. The physical load test was performed by stepping up on to a 30 cm-high stepping stool at a rate of 25/min for 5 minutes, which was a type of moderate physical exercise. The stepping rate and timing were controlled by a metronome.

RESULTSThe levels of DA, E, NE, and ACTH at rest and after physical load were significantly higher in subjects acutely exposed to high altitude than in the sea-level control groups (all P<0.05). Moreover, the levels of DA, E, NE, and ACTH at rest were significantly higher after acute exposure to high altitude in the 5-day HBOP intervention group than in the simulated high-altitude group (all P<0.01). Except for the ACTH level in the 3-day HBOP intervention group, the levels of DA, E, NE, and ACTH after physical load were significantly higher after acute exposure to high altitude in the 3-day and 7-day HBOP intervention groups than in the simulated high-altitude group (all P<0.01).

CONCLUSIONHBOP can elevate the plasma expression of DA, E, NE, and ACTH, and then speed up the establishment of a new balance of homeostasis to adapt to the acute hypoxia at high altitude.

Adrenocorticotropic Hormone ; blood ; Altitude ; Dopamine ; blood ; Epinephrine ; blood ; Exercise ; Homeostasis ; Humans ; Hyperbaric Oxygenation ; Hypoxia ; blood ; Male ; Norepinephrine ; blood ; Rest ; Stress, Physiological

The cross regional loose medical alliance is an important carrier in the current integrated development process of medical services in the Yangtze River Delta region. Smith policy implementation process model was used to analyze the development difficulties of cross regional loose medical alliances from idealized policies, policy implementation institutions, policy target groups, and policy implementation environment. Such medical alliances were formed under the background of integrated development in the Yangtze River Delta, with Shanghai′s tertiary public hospitals as leading units and medical institutions in Jiangsu, Zhejiang, and Anhui provinces as member units. Analysis showed that the policies for such medical alliances development had not yet clearly defined the organizational management mode, operational mechanism, and implementation path, and the corporate governance structure of medical alliance was immature; The policy implementation agencies were relatively lagging behind in the support of special funds and the formulation of related supporting policies; Participation of policy target groups was insufficient and their incentive mechanisms was imperfect; There were problems in the policy implementation environment, namely inconsistent medical and health service regulations and systems in different regions, different health financing capabilities of local governments, insufficient coordination of medical institution management concepts, and a lack of unified standards in information systems. Based on the above difficulties, this study proposed to strengthen the development planning and layout of cross regional loose medical alliances, and improve the corporate governance structure; To strengthen the government′s main responsibility and improving policy implementation capabilities; To improve the internal cooperation and operation mechanism of cross regional loose medical alliances, and enhance the sense of identity of the target group; To optimize the policy implementation environment and implement various support measures, so as to provide references for further promoting the coordinated development of high-quality medical resources in the Yangtze River Delta region.

We proposed a new stitching method based on sift features to obtain an enlarged view of transmission electron microscopic (TEM) images with a high resolution. The sift features were extracted from the images, which were then combined with fitted polynomial correction field to correct the images, followed by image alignment based on the sift features. The image seams at the junction were finally removed by Poisson image editing to achieve seamless stitching, which was validated on 60 local glomerular TEM images with an image alignment error of 62.5 to 187.5 nm. Compared with 3 other stitching methods, the proposed method could effectively reduce image deformation and avoid artifacts to facilitate renal biopsy pathological diagnosis.
Algorithms ; Artifacts ; Humans ; Image Processing, Computer-Assisted ; methods ; Kidney Glomerulus ; ultrastructure ; Microscopy, Electron, Transmission ; methods

Objective To analyze the immunological effect and its influencing factors of HAART on HIV/AIDS with a baseline CD4 of less than 350 cells/mm³ in Hubei Province during 2011-2020. Methods The treatment information of people living with HIV/AIDS (PLWH) was collected through China Disease Control and Prevention Information System, and the change of CD4 mean value was analyzed by Wilcoxon sign rank sum test. Chi square test and multivariate logistic regression were used to analyze the factors influencing the immunological effect. Results The mean value of CD4 cell count at baseline was (172.78±107.65) cells/mm3 before treatment and (398.85±206.71) cells/mm³ in the fifth year after treatment. Patients with baseline CD4<50 /mm3 had the largest increase in CD4 mean value after treatment (P<0.05). Multivariate logistic regression analysis showed that Age ≥35 years, heterosexual transmission, time interval from diagnosis to treatment≥1 month, baseline CD4<50 /mm3, treatment time 7-<18 months, and drug failure were the risk factors for the success of antiviral therapy(P<0.05). Conclusion The immunological effect of HIV/AIDS antiviral therapy in Hubei Province is affected by many factors. It is necessary to strengthen the implementation of early detection and early treatment strategy and promote treatment compliance education for patients aged ≥35 years.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.