Objective:To investigate the relationship between serum retinol binding protein (RBP), stromal cell derived factor-1 (SDF-1) and renal function in patients with diabetic nephropathy (DKD).Methods:The patients with type 2 diabetes mellitus (T2DM) admitted to Yantai Affiliated Hospital of Binzhou Medical College from October 2017 to October 2020 were prospectively selected, 438 patients were divided into simple T2DM group ( n=276)and DKD group( n=162) according to the presence or absence of DKD, according to the ratio of urinary albinin/creatinine (UACR) were divided into normal( n=25), microalbuminuria ( n=75) and macroalbuminuria group ( n=62), according to the estimated glomerular filtration rate (eGFR) were divided into G1 stage ( n=28), G2 stage ( n=27), G3A + G3B stage ( n=35), G4 stage ( n=39)and G5 stages( n=33). The relationship between RBP, SDF-1 and renal function index UACR, serum uric acid (UA), blood urea nitrogen (BUN), β 2-microglobulin (β 2-MG) and serum creatinine (Scr) was analyzed. Measurement data of normal distribution were expressed as Mean± standard deviation ( Mean± SD). Independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups.Chi-square test was used to compare the enumeration data between groups. Receiver operating characteristic curve (ROC) was used to analyze the discriminant value of RBP and SDF-1 for DKD. Pearson was used for correlation analysis among indicators. Multivariate linear regression analysis was used to analyze the influencing factors of RBP. Results:In the DKD group, the duration of diabetes was longer, the levels of RBP, UACR, UA, BUN, β 2-MG, Scr were high, SDF-1 and eGFR were lower, with statistically significant differences compared with the simple T2DM group( P<0.05).The areas under the curve of RBP and SDF-1 to distinguish DKD were 0.903 and 0.868, and the optimal cut-off values was 70.71 mg/L and 5.69 ng/mL. With the increase of urinary albumin and clinical stage, the levels of RBP, UACR, UA, BUN, β 2-MG, Scr increased gradually, while SDF-1 and eGFR decreased gradually, and the differences were statistically significant ( P<0.05).RBP was positively correlated with UACR, UA, BUN, β 2-MG and Scr in DKD patients ( r=0.764, 0.787, 0.693, 0.577, 0.801, P<0.000 1), and negatively correlated with EGFR ( r=-0.782, P<0.000 1). SDF-1 was negatively correlated with UACR, UA, BUN, β 2-MG and Scr ( r=-0.744, -0.794, -0.666, -0.605, -0.820, P<0.000 1), and positively correlated with EGFR ( r=0.767, P<0.000 1). The multiple linear regression equation was RBP=29.852+ 0.007UACR+ 0.101UA+ 0.497BUN+ 0.034Scr-0.083eGFR ( P<0.001). Conclusion:RBP and SDF-1 have certain discriminant value for DKD patients in T2DM population, and the degree of DKD renal function injury is positively correlated with RBP and negatively correlated with SDF-1, the increase of UACR, UA, BUN, Scr and the decrease of eGFR are risk factors for the increase of RBP.

Objective:To analyze the effect of chlorpyrifos on the expression of autophagy related proteins in rat hippocampal neurons, and to explore the role of autophagy in central nerve injury caused by acute chlorpyrifos poisoning.Methods:In October 2018, 35 male clean grade SD rats were randomly divided into 7 groups according to the observation time point, namely 0.5 d, 1 d, 2 d, 3 d, 5 d and 7 d groups and the control group, with 5 rats in each group. Each observation group was given 81.5 mg/kg chlorpyrifos by gavage, and the control group was given olive oil by gavage. The general conditions and poisoning symptoms of rats were observed continuously after exposure. The expressions of autophagy related proteins Beclin1, P62/SQSTM1 and LC3 in hippocampus were detected by Western blot. The cell morphology and LC3 expression in brain were observed by immunohistochemical staining.Results:Western blot results showed that compared with the control group, the expression of Beclin1 protein in hippocampal neurons of rats in the 1 d, 2 d, and 3 d groups increased, while the expression of P62/SQSTM1 protein in the 0.5 d, 1 d, and 2 d groups decreased, and the expression of LC3 protein was decreased in the 2 d group, and the differences were statistically significant ( P<0.05) . The results of immunohistochemistry showed that the hippocampal neurons of rats in the 5 d group were arranged disorderly, and some nuclei contours disappeared, especially in the 7 d group. The LC3 protein was expressed in the cytoplasm, and the expression level gradually increased, reaching a peak on the second day. Conclusion:The early activation of autophagy in rats with acute chlorpyrifos poisoning may be involved in chlorpyrifos induced hippocampal neuronal injury.

Objective:To analyze the effect of chlorpyrifos on the expression of autophagy related proteins in rat hippocampal neurons, and to explore the role of autophagy in central nerve injury caused by acute chlorpyrifos poisoning.Methods:In October 2018, 35 male clean grade SD rats were randomly divided into 7 groups according to the observation time point, namely 0.5 d, 1 d, 2 d, 3 d, 5 d and 7 d groups and the control group, with 5 rats in each group. Each observation group was given 81.5 mg/kg chlorpyrifos by gavage, and the control group was given olive oil by gavage. The general conditions and poisoning symptoms of rats were observed continuously after exposure. The expressions of autophagy related proteins Beclin1, P62/SQSTM1 and LC3 in hippocampus were detected by Western blot. The cell morphology and LC3 expression in brain were observed by immunohistochemical staining.Results:Western blot results showed that compared with the control group, the expression of Beclin1 protein in hippocampal neurons of rats in the 1 d, 2 d, and 3 d groups increased, while the expression of P62/SQSTM1 protein in the 0.5 d, 1 d, and 2 d groups decreased, and the expression of LC3 protein was decreased in the 2 d group, and the differences were statistically significant ( P<0.05) . The results of immunohistochemistry showed that the hippocampal neurons of rats in the 5 d group were arranged disorderly, and some nuclei contours disappeared, especially in the 7 d group. The LC3 protein was expressed in the cytoplasm, and the expression level gradually increased, reaching a peak on the second day. Conclusion:The early activation of autophagy in rats with acute chlorpyrifos poisoning may be involved in chlorpyrifos induced hippocampal neuronal injury.

Objective: To establish the Wistar rat model of acute diquat poisoning and observe the pathological damage of main target organs. Methods: Thirty-six Wistar rats were randomly divided into six groups (n=6) , including one normal saline control group and five treatment groups which were separately given single-dose of intragastric administration at the doses of 46.2 mg/kg, 77.0 mg/kg, 115.5 mg/kg, 231.0 mg/kg and 346.5 mg/kg. The pathological changes of lung, liver and kidney were observed by hematoxylin and eosin (HE) and Masson staining. The optimal dose was determined according to the general situation and pathological changes. Thirty-six Wistar rats were randomly divided into five treatment groups and one normal saline control group. Treatment groups were given single-dose of intragastric administration according to the optimal dose. The rats were sacrificed at 1st, 3rd, 7th, 11th and 14th day after exposed, respectively. The activity of serum glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) were measured by chemical colorimetry. The pathological changes of lung, liver and kidney were observed by HE and Masson staining. Results: According to 14 d survival rate, the toxic symptoms and pathological changes, 115.50 mg/kg was determined the best dose. Given single-dose of intragastric administration at the doses of 115.50 mg/kg, it was found that the serum AST and ALT activity of rats on the first and third day of exposure was significant higher than those in control group. The results of pathological examination exhibited that in 115.50 mg/kg group, the pathological changes of lung, liver and kidney began to appear on the first day of exposure, the pathological changes were the most serious on the third day, and then gradually alleviated. On the 14th day, the alveolar septum was slightly widened, with inflammatory cell infiltration, local alveolar cavity became narrow, atrophy, peripheral alveolar compensation, bronchi and alveolar septum collagen fiber proliferation; The local renal tubular epithelial cells were enlarged and necrotic; the central vein surrounding hepatic cells showed vacuolar degeneration with punctate necrosis. Conclusion The rat model of acute diquat poisoning can be successfully induced by single-dose of intragastric administration. The condition of wistar rats and the pathological damage of the main target organs could be observed during the whole course of 115.50 mg/kg administration.

Objective: To investigate the clinical and laboratory features of Aeromonas bacteremia in patients with hematological diseases, and provide evidence for the prevention and treatment of Aeromonas infection. Methods: A retrospective study of patients with bloodstream infection of Aeromonas in our hospital from January 2014 to December 2018 was carried out. The clinical characteristics, antimicrobial susceptibility, infection seasons, antimicrobial therapy and evolution were analyzed. Results: A total of 42 patients with hematological diseases had Aeromonas bloodstream infection within 5 years. Among them, 39 cases (92.9%) of bloodstream infection occurred in the stage of neutropenia. The median time of fever was 4 (1-27) d, 22 (52.4%) patients only had fever, 6 (14.3%) with gastrointestinal symptoms (abdominal pain, diarrhea, nausea, upper gastrointestinal bleeding) , 8 (19.0%) with pulmonary infection, 13 (31.0%) with skin and soft tissue infections. Seven patients (16.7%) died with skin and soft tissue infection. The resistance of Aeromonas to carbapenems was 68.3%-70.7%, while the resistance rate to cephalosporins, quinolones and aminoglycosides were less than 10%. Conclusions Aeromonas bacteremia in patients with hematological diseases mainly occur in the neutropenia stage, usually with symptom like fever. The mortality is increased when accompanied by skin and soft tissue infection. Antibiotic use should be based on susceptibility results, and avoid the use of carbapenems.

Objective To study the pathological changes of the sudden death cases due to the dysplasia of the cardiac conduction system. Methods Using the newly established pathological sampling method by our research team, 26 hearts of unidentified sudden deaths excluding trauma, poisoning and diseases were collected and observed optically with H&E staining, Masson staining and immunohistochemical staining. Results Among the 26 cases, there were dysplasia in the cardiac conduction system,structural abnormalities, tissue or structure shift, fibrous cushion in the sinoatrial node and 9 cases have 2~3 abnormalities at the same time. Among the age groups from 16 to 30, 21 cases (80.8%) showed dysplasia of the cardiac conduction system. Conclusion The dysplasia of the cardiac conduction system may be one of the predominant reasons for the sudden death in young adults.

BACKGROUND:Epithelial cel s are commonly used as the seed cel in tissue engineering;however, there is stil a lack of an effective in vivo noninvasive trace technology. OBJECTIVE:To investigate the feasibility of labeling canine oral epithelial cel s with ultrasmal superparamagnetie iron oxide (USPIO) and magnetic resonance imaging (MRI) in vitro. METHODS:Oral epithelial cel s from beagles were primary cultured, and then labeled by 0.75 mg/L poly-L-lysine combined with USPIO (0, 5, 10, 25, 50 and 100 mg/L), respectively. To determine the optimal dosage, the intracel ular iron expression was identified by Prussian blue staining, and the cel viability in different groups was detected by cel counting kit-8. Final y, 2×105 labeled cel s were suspended with 1 mL PBS buffer, and were screened using 3.0 T MR on T2*WI sequences in vitro. RESULTS AND CONCLUSION:USPIO prepared with 0.75 mg/L poly-L-lysine could successful y label dog oral epithelial cel s. Prussian blue staining showed intracel ular blue spots, and the intracel ular blue spots became more with the concentration increasing and saturated at the concentration of 25 mg/L. Cel counting kit-8 indicated that the cel viability did not change when the concentration<25 mg/L. Among the T2*WI sequences, the MRI signal intensity decreased with the concentration increasing. In conclusion, canine oral epithelial cel s can be effectively labeled with USPIO making no impact on cel viability when the concentration<25 mg/L, and MRI can be used to track these labeled cel s in vitro.

Tumors of the cardiac conduction system (CCS) have rarely been reported. The CCS from 198 cardiac-related deaths (GroupⅠ),and 838 deaths from non-cardiovascular diseases or trauma (Group Ⅱ ), were studied. Sampling was done of the sinoatrial node (SAN)and atrio-ventricular node (AVN) along their long axis of each node as a single block and the His bundle (HB) perpendicular to its long axis in 2-4 blocks. Five-micron serial sections were made; tissue slices were taken intermittently, every 20th from the SAN, every 10th from the AVN, and every 30th from the HB and bundle branches (BB), by continuous slices three times. Tumors in the CCS were found in 12 cases (1.155 %), where 10 (0.965%) were primary tumors, and 2 (0.193%) were metastatic tumors. The primary tumors included 4 fibromata compressing the HB (0.386 %), 4 hemangiomata (0.386%), 1 AVN tumor (0.097 %), and 1 rhabdomyoma (0.097 %). In 8 of the 10 cases, the tumors were located in the AVN or HB. The metastatic tumors originated from lymphocytic leukemia and malignant lymphoma (histiocytic type) in lung, and were all found in the SAN. Of the 12 cases, 2 were from the group Ⅰ. Tumors in the CCS are the smallest tumors in different parts of the body, which can cause sudden death.

Objective To study the pathological changes related to cardiac sudden death. Methods The CCS from 179 cases with cardiac sudden death were examined histopathologically. Result There were 8 cases with hemorrhage, inflammation, or tumor infiltration in the cardiac nervous tissue or cardiac ganglion. Pathological changes such as edema, degeneration or necrosis could be found in the nervous tissue. However, no lethal pathological changes were observed out of cardiac nervous tissue. Conclusion Either primary or sencondary pathological changes in the cardiac nervous tissue might cause sudden death.

Objective To study the ultrastructural changes of gunshot wounds at the brain stem and the cause of the death. Method A case died of the head injury and two cases died of gunshot wounds of the brain stem were studied. Brain stem tissues were taken 25 min after death. Tissue blocks from the tegmentum of the midbrain and the pons as well as the gray matter of the medulla oblongata in the deep portion of the base of the fourth ventricle were taken and were examined with ultramicroscopy and scanning electron microscopy. Results (1) The gunshot wound injuries inside the brain including burn and injuries caused by punch and pressure; (2) Although the primary gunshot wounds were located at the midbrain, the pathological changes extended to the whole brain stem; (3)The injuries of the axons and the nerve cells were more severe and extensive than those of the glia fibers. (4)The changes of the burn included condensation and degeneration of the myelin sheath and axonal cytoplasm, especially the mitochondria as well as edema of the myelin sheath and the axonal cytoplasm. The punch and pressure injuries included the torsion, deformation, fusion, compression, breach, torn and displacement of the axons with loss of cytoplasm as well as the deviation of mitochondria, micro ?neuro - fibrils and micro - tubules in the axons. Conclusion The severe and extensive injuries of the axons of nerve fibers in the brain stem might play an important role in the cause of death of gunshot.