Objective To investigate the regulation of melatonin (MT) on Toll-like receptor 4 (TLR4) signaling in diabetic db/db mice kidneys.Methods The 48 10-week-old male db/db mice were randomly divided into db/db group,db/db+MT 50 μg/kg group,db/db+MT 100 μg/kg group and db/db+MT 200 μg/kg group,each consisting of 12 mice.These mice received i.p.injections of MT These mice received i.p.injections of MT [dissoved in phosphate buffer solution (PBS)/ dimethylsulfoxide (DMSO) solution,given every day].Alternatively,12 db/m mice served as the control group.db/m and db/db group were injected i.p.with the same volume of PBS/DMSO solution.The animals were sacrificed after 12 weeks of dosage administration.Blood glucose (BG),body weight (BW),kidney weight (KW) and 24 h urinary albumin excretion rate (UAER) were determined;Kidney pathological lesions were evaluated by renal pathological staining.Immunohistochemistry of renal TLR4,NF-κB p65,and ED-1 was performed to determine the immunoreactivity.Western blotting was used to detect the expression of renal TLR4,myeloid differentiation factor 88 (MyD88),TIR-domaincontaining adaptor inducing interferon-β (TRIF),interferon regulatory factor 3 (IRF-3) and NF-κB p65,while the mRNA expressions of renal tumor necrosis factor-α (TNF-α) and monocyte chemotactic protein-1 (MCP-1) were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were much higher in db/db mice group (P ＜ 0.01),while KW in db/db+MT (100,200 μg/kg) groups and UAER level in db/db+MT (50,100,200 μg/kg) groups were distinctly decreased compared with those in db/db group (P ＜ 0.01).In week 12 db/db mice,the glomerular mesangial expansion index and tubulointerstitial injury index were increased compared with those in db/m mice (P ＜ 0.01).The above kidney histopathologic lesions were distinctly ameliorated by 50,100,200 μg/kg MT (P ＜ 0.05).Immunohistochemistry intensity of renal TLR4,NF-κB p65 and ED-1 displayed obvious differences between db/m mice and db/db mice (P ＜ 0.01),and that were remarkably decreased in db/db+MT (50,100,200 μg/kg) mice compared with db/db mice (P ＜ 0.05).Western blotting showed that the protein expression of renal TLR4,MyD88,TRIF,IRF-3 and NF-κB p65 were stronger in db/db group compared with those in db/m group (P ＜ 0.05) and weaker in db/db+ MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.05).Futhermore,the mRNA expressions of renal MCP-1 and TNF-α were higher in db/db group compared with those in db/m group (P ＜ 0.01) and lower in db/db+MT (50,100,200 μg/kg) groups compared with those in db/db group (P ＜ 0.01).Conclusion Melatonin may partly down-regulate TLR4 signaling pathway to inhibit Inflammatory reaction and alleviate kidney injury in diabetic db/db mice.

Objective To investigate the efficacy of preemptive analgesiawith ultrasound-guided modified dorsal penile nerve block combined ketorolac tromethamine for circumcision in pediatric patients.Methods Forty-five pediatric patients with ASA grade Ⅰ undergoing elective circumcision were randomly divided into 3 groups (n=15 each).The same intravenous compound anesthesia was used in the three groups.The pediatric patients underwent ultrasound-guided modified dorsal penile nerve block with a mixture of 0.1 ml/kg of 0.2% ropivacaine and 0.8% lidocaine before operation in group A and group B.The pediatric patients were given ketorolac tromethamine 1 mg/kg intramuscular injection before operation in group A and group C.The total consumption of propfol and sufentanil, occurrence of intraoperative body movement and respiratory depression, emergence time, time from waking up to going out of PACU and adverse reactions such as postoperative agitation, nausea, vomiting and pruritus were aslo recorded.The requirement for postoperative paracetamol suppositories was recorded.Results Compared with group C, the total consumption of propfol and sufentanil were significantly decreased, incidence of body movement andrespiratory depression were significantly decreased, the emergence time and time from waking up to going out of PACU was significantly shortened, requirement for paracetamol suppositories were significantly decreased in group A and group B(P<0.05).Compared with group B, incidence of requirement for paracetamol suppositories was significantly decreased in group A (P<0.05).No pediatric patients developed postoperative nausea, vomiting, pruritus and incidence of emergence agitation had no statistical difference in the three groups.Conclusion Preemptive analgesia with ultrasound-guided modified dorsal penile nerve block combined ketorolac tromethamine is safe and effective when used for circumcision in pediatric patients, and it has good efficacy of postoperative analgesia.

Objective To explore the relationship among genetic polymorphism of angiotension Ⅱ type 1 re-ceptor(AT1 R) A1166-C, angiotensin converting enzyme (ACE) insertion/deletion (I/D), aldosterone synthase (CYP11B2)-344T/C and hypertensive disorder complicating pregnancy.Methods Polymerase chain reaction-re-striction fragment length polymorphism (PCR-RFLP) assay was used to detect the genotypes of AT1 R A1166-C ,ACE (I/O) ,CYP11B2 -344T/C in 86 cases of hypertensive disorder complicating pregnancy and 175 cases of normal control.Results There was 18 combined types in hypertensive disorder complicating pregnancy cases and normal control cases.Compared to AT1R-AA + ACE-Ⅱ + CYP11B2-TT, Odds ratios (OR) of AT1R-AA + ACE-DO +CYP11B2-TC,AT1 R-AC + ACE-ID+CYP11B2-TC and AT1R-AC+ACE-DD+CYP11B2-TC are 7.289,5.315 and 5.694 respectively.There was no statistical significance among the others.Conclusion In all 18 kinds of combined types, AT1 R-AA + ACE-DO + CYP11B2-TC,AT1R-AC+ACE-ID+CYP11B2-TC and AT1 R-AC + ACE-DD +CYP11B2-TC might increase the susceptibility of hypertensive disorder complicating pregnancy.It is possible that multigenes are interacted in the etiology of hypertensive disorder complicating pregnancy.

Objective:To explore the role of hippocampal γ oscillation abnormality in sepsis-associated encephalopathy (SAE).Methods:Seventy male Sprague-Dawley rats (2-3 months) were randomly (random number) divided into three groups according to the random digital table method: sham, CLP, and CLP + dopamine 4 (D4) receptor agonists RO-10-5824 group. The SAE animal model was established by cecal ligation and puncture (CLP). On day 10-14 after surgery, the open field, novel object recognition, and fear conditioning tests were performed. After that, the hippocampus was collected to measure expressions of parvalbumin (PV) and D4 receptor. In another set of experiment, CA1 local field potential (LFP) were recorded, and the relationship between LFP and time with novel object was analyzed. Independent sample t-test was used for pairwise comparisons, and multiple comparisons were performed by one-way ANOVA, followed by the Tukey multiple comparisons test. Correlation was analyzed using Pearson correlation. Statistical significance was assumed when P<0.05. Results:Compared with the sham group, hippocampal PV (77.54±4.61)%, D4 expression (56.36±3.88)% and γ oscillation power (41.1±8.62)%, object exposure time (36±3) s, new object recognition rate (49±4)%, and scene stiffness time (56±7) s were decreased significantly ( P<0.05). However, RO-10-5824 treatment could increase hippocaml γ oscillation power (92.3±6.7)%, and reverse the decreased new object exposure time (44±3) s and new object recognition rate (63±4)%. Correlation analysis showed that hippocampal γ oscillation power was positively associated with new object exposure time ( r=0.609 2, P=0.015 9). There was no difference in total distance traveled or time spent in the center among groups ( P>0.05). Conclusion:Hippocampal γ oscillation abnormality might play a key role in cognitive impairment associated with SAE.

Peak Torque(PT),Total Work(TW),Average Power(AP),Torque Acceleration Energy(TAE),Set Total Work(STW),Endurance Ratio (ER)and DOrsal/plantar ratio of ankle dorsal and plantarflexion were tested in 101 patients with lumbar disc protrusion using a Cybex一6000 isokinetic muscle test-ing instrument.The results showed the most tested of the affected side was signficantly lower than that onthe opposite side. The PT,TW and STW of the affected side was lower 70一80%than that on the oppo-site side. The rest was lower 20一30%than another side. No significant difference of endurance ratios be-tween the affected and unaffected sides was noted.

Objective To study the regulation of microRNA - 22(miR - 22)on glycometabolism of hemato-poietic stem cell TF - 1 and its molecular mechanism. Methods TF - 1 cells were cultured for 2 h under hypoxic con-ditions. The expression levels of Glut4 and miR - 22 was detected by quantitative real-time PCR(qRT - PCR). The sgRNA vector of the miR - 22 gene was constructed and miR - 22 gene of TK - 1 cells was knocked out by the CRISPR/ Cas9 technique. Overexpression vectors were constructed,and miR - 22 knocked - out cells were introduced to overexpress miR - 22,the expression of miR - 22 was detected by qRT - PCR and the expression levels of Glut4 and PPAR - γ were detected by qRT - PCR and Western blot. Results Compared with the control group,the expression of miR - 22 in TF - 1 cells decreased (0. 015 ± 0. 000 vs. 0. 056 ± 0. 001)and the expression of Glut4 (0. 351 ± 0. 038 vs. 0. 152 ± 0. 005)and PPAR - γ (0. 421 ± 0. 017 vs. 0. 248 ± 0. 008)increased,when TF - 1 cells were cultured for 2 h under hypoxic conditions,and the differences were statistically significant (all P < 0. 05). Compared with the control group,the expression levels of Glut4 (0. 019 ± 0. 00 vs. 0. 008 ± 0. 000)and PPAR - γ (0. 038 ± 0. 001 vs. 0. 019 ± 0. 000)were significantly increased after miR - 22 gene silencing,and they were significantly decreased (Glut4:0. 005 ± 0. 000 vs. 0. 008 ± 0. 000;PPAR - γ:0. 137 ± 0. 000 vs. 0. 019 ± 0. 000)after overexpression of miR - 22,and the differences were statistically significant (all P < 0. 05). Conclusions It suggests that miR - 22 ex-erts a negative regulation on glycometabolism of hematopoietic stem cell TF - 1 by downregulating the expression of PPAR - γ. A new regulatory factor of TF - 1 glycometabolism and the mechanisms are identified,which has provided new ideas for the targeted medication of diseases induced by hematopoietic stem cell dysfunction.

Objective To investigate the effects of Nod-like receptor protein 3 inhibitor MCC950 on cognitive function in a mouse model of sepsis-associated encephalopathy (SAE).Methods Ninety adult male C57BL/6 mice were randomly (random number) divided into three groups:the sham + saline group (n=20,sham group),CLP + saline group (n=35,CLP group),and CLP + MCC950 group (n=35,MCC950 group).SAE mouse model was established by cecal ligation and puncture (CLP) surgery.Saline (10 mL/kg) or MCC950 (10 mg/kg) was intraperitoneally injected 30 min before surgery and on day 1,2,4 and 6 after surgery according the grouping.Seven days after surgery,six mice were taken from each group.Western blot was used to detect the hippocampal content of NLRP3,apoptosis-associated specklike protein (ASC),interleukin-1β (IL-1β) and IL-18.The number of NLRP3-positive cells in CA 1 region were detected by immunohistochemical analysis.The remaining mice in each group were used for open field and fear conditioning tests 14 days after surgery.One-way analysis of variance was used for inter-group comparison,and SNK-q test was used for pairwise comparison.A P value ＜0.05 was considered statistically significant.Results Compared the MCC950 group with the CLP group,the freezing time of context test was significantly increased [(137±21) s vs (84±15) s,P=0.013],the hippocampal content of NLRP3,IL-1β and IL-18 were significantly reduced (P＜0.01),and the number of NLRP3-immunoreactive cells per mm2 in CA region were significantly decreased (23±5 vs 74±13,P＜0.01).There was no significant changes in protein level of ASC and results of open field tests (P＞0.05).Conclusions MCC950 administration can improve cognitive function in a mouse model of SAE,which is probably due to the inhibition of NLRP3 inflammasome and downstream inflammatory cytokines IL-1β and IL-18.

Objective To evaluate the effect of methylene blue(MB)preconditioning on ischemi-a-reperfusion(I∕R)injury in isolated rat lungs. Methods Eighteen pathogen-free healthy male Sprague-Dawley rats, aged 3 months, weighing 240-320 g, were divided into 3 groups(n=6 each)using a ran-dom number table: sham operation group(group Sham), lung I∕R group(group I∕R)and methylene blue preconditioning group(group MB). A model of isolated lung I∕R injury was established in pentobarbi-tal sodium-anesthetized rats. MB 2 mg∕kg was intraperitoneally injected at 2 h before stopping perfusion in group MB. Isolated lungs were perfused for 20 min, followed by 45-min ischemia, and then reperfused for 60 min in I∕R and MB groups. At 60 min of reperfusion, the activity of lactic dehydrogenase(LDH)in the perfusate was detected, wet weight(W)and dry weight(D)was determined, W∕D ratio was calcu-lated, and the levels of malondialdehyde(MDA), ATP, reactive oxygen species(ROS)and superoxide dismutase(SOD)were measured in lung tissues. Mitochondria and cytoplasm were isolated from lung tis-sues for determination of mitochondrial membrane potential(MMP), degree of mitochondrial swelling and content of cytochrome C(Cyt c)in cytoplasm. Apoptotic cells in lung tissues were detected using TUNEL, and apoptotic index was calculated. Results Compared with group Sham, the activity of LDH in perfu-sate, W∕D ratio, levels of ROS, MDA and Cyt c in cytoplasm and apoptosis index were significantly in-creased, the degree of mitochondrial swelling was aggravated, and the content of ATP and MMP were de-creased in I∕R and MB groups, and the SOD activity was significantly decreased in group I∕R(P<005). Compared with group I∕R, the activity of LDH in perfusate, W∕D ratio, levels of ROS, MDA and Cyt c in cytoplasm and apoptosis index were significantly decreased, the degree of mitochondrial swelling was attenu-ated, and the activity of SOD, content of ATP and MMP were increased in group MB(P<005). Con-clusion Methylene blue preconditioning can reduce I∕R injury in isolated rat lungs, and the mechanism may be related to improving mitochondrial function and inhibiting cell apoptosis.

Objective To analyze the clinical risk factors for chronic complications in patients with type 2 diabetes and their correlation with bone mineral density and 1,25-dihydroxyvitamin D3. Methods A total of 163 patients with type 2 diabetes mellitus were selected as research subjects and were divided into complication group and non-complication group according to the presence or absence of chronic complications. The independent related factors for chronic complications in patients with type 2 diabetes mellitus were analyzed. Spearman rank correlation analysis was used to evaluate the correlation between bone mineral density, 1,25-dihydroxyvitamin D3 and chronic complications. Results Among the 326 patients with type 2 diabetes mellitus, 202 developed chronic complications (61.96%), including 71 cases of cardiovascular disease, 59 cases of neuropathy, 33 cases of renal lesion, and 28 cases of retinopathy. There were statistically significant differences in the duration of diabetes mellitus, fasting blood glucose, systolic blood pressure, glycosylated hemoglobin, triglyceride, low density lipoprotein cholesterol, serum creatinine, bone mineral density, and 1,25-dihydroxyvitamin D3 between the complication group and the non-complication group (P<0.05). Logistic multivariate regression analysis showed that the duration of diabetes mellitus, systolic blood pressure, glycosylated hemoglobin, ow density lipoprotein cholesterol, serum creatinine, bone mineral density, and 1,25-dihydroxyvitamin D3 were all independent related factors for the occurrence of chronic complications in patients with type 2 diabetes mellitus (P<0.05). Spearman correlation analysis showed that bone mineral density and 1,25-dihydroxyvitamin D3 were negatively correlated with chronic complications (P<0.05). Conclusion Bone mineral density and 1,25-dihydroxyvitamin D3 in patients with type 2 diabetes mellitus are closely related to chronic complications.

Objective To compare the conjoint effect of enteral nutrition (EN) and parenteral nutrition (PN) with single EN or PN on immune function, nutritional status, complications and clinical outcomes of patients with severe traumatic brain injury (STBI). Methods A prospective randomized control trial was carried out from January 2009 to May 2012 in Neurological Intensive Care Unit (NICU). Patients of STBI who met the enrolment criteria (Glasgow Coma Scale score 6~8; Nutritional Risk Screening ≥3) were randomly divided into 3 groups and were admi- nistrated EN, PN or EN+PN treatments respectively. The indexes of nutritional status, immune function, complications and clinical outcomes were examined and compared statistically. Results There were 120 patients enrolled in the study, with 40 pationts in each group. In EN+PN group, T lymthocyte subsets CD3+%, CD4+%, ratio of CD3+/CD25+, ratio of CD4+/CD8+, the plasma levels of IgA, IgM, and IgG at 20 days after nutritional treatment were significantly increased compared to the baseline(t=4.32-30.00, P<0.01), and they were significantly higher than those of PN group (t=2.44-14.70; P<0.05,or P<0.01) with exception of CD4+/CD8+, higher than those of EN group (t=2.49-13.31, P<0.05, or P<0.01) with exceptions of CD3+/CD25+, CD4+/CD8+, IgG and IgM. For the nutritional status, the serum total protein, albumin, prealbumin and hemoglobin were significantly higher in the EN (t=5.87-11.91; P<0.01) and EN+PN groups (t=6.12-13.12; P<0.01) than those in PN group after nutrition treatment. The serum prealbumin was higher in EN+PN group than that in EN group (t=2.08; P<0.05). Compared to the PN group, the complication occurrence rates of EN+PN group were significantly lower in stress ulcer (22.5% vs. 47.5%; χ= 8.24, P<0.01), intracranial infection (12.5% vs 32.5%;χ= 6.88, P<0.01) and pyemia (25.0% vs. 47.5%; χ= 6.57, P<0.05). Compared to the EN group, the complication occurrence rates of EN+PN group were significantly lower in aspirated pneumonia (27.5% vs. 50.0%; χ= 6.39, P<0.05), hypoproteinemia (17.5% vs. 55.0%; χ= 18.26, P<0.01) and diarrhea (20.0% vs. 60.0%; χ= 20.00, P<0.01). The EN+PN group also had significant less length of stay in NICU (t=2.51, 4.82; P<0.05, P<0.01), number of patients receiving assisted mechanical ventilation (χ= 6.08, 12.88; P<0.05, P<0.01) and its durations (t=3.41, 9.08; P<0.05, P<0.01), and the death rate (χ=7.50, 16.37; P<0.05, P<0.01) than those of EN or PN group. Conclusion Early EN+PN treatment could promote the recovery of the immune function, enhance nutritional status, decrease complications and improve the clinical outcomes in patients with severe traumatic brain injury.
Adult ; Brain Injuries, Traumatic ; immunology ; metabolism ; therapy ; Enteral Nutrition ; Female ; Humans ; Male ; Middle Aged ; Nutritional Status ; Parenteral Nutrition ; Treatment Outcome