Objective To investigate the protective effects and mechanism of insulin(INS) on brain in septic rats,and explore the possible role of uncoupling protein 2 (UCP2) in these effects.Methods Fifty male specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into normal control (CN) group(n=10),lipopolysaccharide(LPS) group(n=20) and INS group (n=20) according to random number table.The septic rat model was established through an intraperitoneal injection of 15 mg/kg LPS of gram-negative bacteria.The rats in the INS group received a 1 U/kg INS injection subcutaneously 30 minutes before the injection of LPS,and the rats in the CN group were given equivalent 9 g/L saline in the same way.Eight rats in each group were killed,and their cerebral cortex were collected after the injection of LPS for 24 h.Pathological change of cerebral cortex was detected by Hematoxylin-Eosin(HE) staining.The cerebral cortex mitochondia were extracted for detecting the levels of reactive oxygen species(ROS),malondialdehyde (MDA) and the activity of superoxide dismutase(SOD).Neuronal apoptosis was detected by terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling staining.UCP2 mRNA expression was detected by quantitative real-time(RT)-PCR.Apoptosis-associated protein B lymphocyte tumor-2(Bcl-2),Bcl-2 associated X protein(Bax),cleaved cysteinyl aspartate specific protease(cleaved Caspase-9) and UCP2 protein expression were determined by Western blot.Results (1)Compared with the CN group,obvious abnormal pathological change was revealed by HE staining in cerebral cortex of rats in the LPS group and the INS group,but the pathological change was attenuated in the INS group compared with the LPS group.(2)Compared with the CN group,the levels of mitochondrial ROS[(210.01±14.09) RFU vs.(49.06±7.28) RFU] and MDA[(2.19±0.18) nmol/mg pro vs.(1.25±0.11)nmol/mg pro]in the LPS group significantly increased,whereas SOD activity significantly decreased [(238.49±35.60) U/g pro vs.(446.66±24.90)U/g pro],and the differences were significant(all P<0.05).Compared with the LPS group,the levels of ROS [(152.69±15.83) RFU vs.(210.01±14.09) RFU] and MDA[(1.55±0.14) nmol/mg pro vs.(2.19±0.18) nmol/mg pro] in the INS group decreased,while SOD activity increased[(327.8±23.26) U/g pro vs.(238.49± 35.60) U/g pro],and the differences were significant(all P<0.05).(3)Compared with the CN group,the neuronal apoptosis index of cortex in the LPS group was elevated[(54.16±6.84)% vs.(5.45±1.43)%],while the expression of Bcl-2 decreased (627±0.018 vs.0.739±0.020),but the expressions of Bax(0.768±0.019 vs.0.520±0.010) and cleaved Caspase-9(0.739±0.016 vs.0.467±0.030) increased,and the differences were significant(all P<0.05).Compared with the LPS group,the neuronal apoptosis index of cortex in the INS group decreased [(33.30±3.07)% vs.(54.16±6.84)%],but the Bcl-2 expression increased (0.743±0.022 vs.0.627±0.018),and Bax (0.687±0.034 vs.0.768±0.019) and cleaved Caspase-9(0.551±0.013 vs.0.739±0.016) were reduced,and the differences were significant (all P<0.05).(4)Compared with the CN group,the mRNA (2.248±0.155 vs.1.000±0.100) and protein expression of UCP2 (0.659±0.016 vs.0.599±0.018) were elevated in the LPS group.Compared with the LPS group,the UCP2 mRNA (2.944±0.117 vs.2.248±0.155) and UCP2 protein (0.719±0.018 vs.0.659±0.016) increased,and the differences were significant(all P<0.05).Conclusions INS can protect the brain of septic rats through alleviating mitochondrial oxidative stress and inhibiting the mitochondrial-initiated apoptotic pathway to reduce neuronal apoptosis.INS upregulates UCP2 expression in the brain of septic rats,which may play a role in the protective effects mentioned above.

OBJECTIVETo observe the relationship between the contents of interleukin-1beta (IL-1beta), interleukin-4 (IL-4) and interleukin-10 (IL-10) in the seminal plasma of infertile males and sperm function indexes.

METHODSBy radioimmunoassay (RIA), we determined the contents of IL-1beta, IL-4 and IL-10 in the seminal plasma of 126 infertile and 20 normal males. According to the sperm count, the infertile were divided into three groups: Groups A (sperm count > or = 20 x 10(6)/ml), B (sperm count < 20 x 10(6)/ml) and C (azoospermia). Based on sperm vitality and motility, Group A was subdivided into a normal and abnormal vitality group and a normal and decreased motility group. In line with the serum results of antisperm antibody (AsAb) and semen WBC, the infertile males were divided into AsAb positive and negative, and WBC semen and non-WBC semen groups. According to the assay results of normal males, Groups A and B were each subdivided into normal and decreased groups of sperm penetrating power, intact acrosome rate and terminal swelling rate.

RESULTSThe content of IL-1beta in the seminal plasma of the infertility group was obviously higher, but the content of IL-4, IL-10 significantly lower than that of the normal group (P < 0.01). In the infertility group, there existed significant differences in the contents of IL-1beta, IL-4, IL-10 in seminal plasma between the WBC and non-WBC semen groups, as well as between the AsAb positive and negative groups (P < 0.05 or P < 0.01); and the same was true for the content of IL-4 between the normal and decreased groups of sperm vitality, motility, penetrating power, intact acrosome rate, and terminal swelling rate (P < 0.05 or P < 0.01).

CONCLUSIONThe contents of IL-1beta, IL-4 and IL-10 in seminal plasma are closely related to male reproduction. The increase or decrease of the contents reflects the state of immunity and infection of the reproductive system, and influences sperm functions.

Adult ; Case-Control Studies ; Female ; Humans ; Infertility, Male ; immunology ; metabolism ; Interleukin-1 ; analysis ; Interleukin-10 ; analysis ; Interleukin-4 ; analysis ; Male ; Radioimmunoassay ; Semen ; chemistry ; Sperm Count ; Sperm Motility

BACKGROUND:Cordyceps polysaccharide is a commonly used adjuvant drug for clinical treatment of nephrotic syndrome. As a classic model of nephrotic syndrome induced by adriamycin, the Sprague-Dawley rat model of nephrotic syndrome exhibits similar clinical manifestations and pathological changes to minimal-change nephropathy in humans. OBJECTIVE:To observe the effect of adipose-derived mesenchymal stem cel s (ADMSCs) transplantation combined with cordyceps polysaccharide on renal function and hypercoagulable state in rats with nephrotic syndrome. METHODS:ADMSCs suspension was made in vitro and labeled using PKH26. Fifty Sprague-Dawley rats were randomized into normal (no intervention), model, ADMSCs, cordyceps polysaccharide and combined treatment groups (n=10/group). Adriamycin administration was performed to make rat models of nephrotic syndrome in the latter three groups. After modeling, model rats were respectively given no treatment, ADMSCs intravenously for 3 days, cordyceps polysaccharide intragastrical y for 12 weeks, or their combined use. Then, 24-hour urinary protein, serum total cholesterol, triglyceride, total protein, albumin, high-density lipoprotein, low-density lipoprotein, creatinine, blood urea nitrogen levels and coagulation changes were detected at 12 weeks. Meanwhile, histopathological changes of renal tissues were observed under light microscope;survival and distribution of PKH26-labeled ADMSCs were observed by fluorescence microscopy;and expression of Hpa gene in renal tissue was detected by RT-PCR. RESULTS AND CONCLUSION:Compared with the model group, the 24-hour urinary protein, serum total cholesterol, triglyceride, low-density lipoprotein, creatinine, and blood urea nitrogen levels were significantly lower, while the serum total protein, albumin and high-density lipoprotein levels were significantly higher in the three treatment groups (P<0.05). These indicators showed significant differences between the combined group and ADMSCs and cordyceps polysaccharide groups (P<0.05). Both ADMSCs transplantation and cordyceps polysaccharide significantly relieved the hypercoagulable state of rats with nephrotic syndrome, and their combined effects were stronger (P<0.05). After treatment, the pathological improvement in the kidney tissues was found in the three treatment groups;moreover, it was better in the combined treatment group than the ADMSCs and cordyceps polysaccharide groups. Better improvement in the number of PKH26-labeled ADMSCs and the expression of Hpa mRNA was observed in the combined treatment group compared with the ADMSCs and cordyceps polysaccharide groups. In conclusion, the combination of ADMSCs transplantation and cordyceps polysaccharide can improve kidney function and hypercoagulable state in rats with nephrotic syndrome, reducing pathological damage to the kidney tissue.

Objective:To investigate the efficacy and feasibility of radiofrequency resection of supraglottic laryngeal carcinoma under a multifunctional opener.Methods:The clinical data of four cases of supraglottic laryngeal carcinoma (type T1N1M0 in two cases, T2N1M0 in one case, and T1N0M0 in one case) who received treatment in the Affiliated Hospital of Jining Medical University during January-June 2019 were retrospectively analyzed. Radiofrequency resection of supraglottic laryngeal carcinoma under a multi-functional opener combined with bilateral neck II-IV region lymph node dissection was performed. Swallowing, breathing and phonation were observed and analyzed based on references.Results:Among the four cases, two cases had a normal diet at 3 days after surgery, one case had a normal diet at 7 days after surgery, and one case had a normal diet at 16 days after surgery. Tracheotomy was not performed in any case. After surgery, breathing and speech communication were not affected.Conclusion:Radiofrequency surgery under a multifunctional opener can be used for treatment of early supraglottic laryngeal cancer. It is an effective treatment with minimal trauma, mild postoperative pain and promotes the early recovery of normal swallowing function.

Objective To preliminarily investigate the role of Vitamin C in cecal ligation and puncture-induced septic brain injury.Methods Male specific pathogen free (SPF) Sprague-Dawley male rats were randomly assigned into control group,sham operation group,sepsis group and sepsis therapy group.The rats in sepsis group were prepared by cecal ligation and puncture.The rats in sepsis therapy group were injected sodium ascorbate through the tail vein 3 h after the cecal ligation and punature procedure.The animals in other groups were subjected only to subcutaneous bolus injection of 9 g/L saline only.Animals were evaluated by neurologic reflex scores before sacrifice and brain tissues were quickly removed at the indicated time points.Reactive oxygen species (ROS),superoxide dismutase (SOD),malondialdehyde (MDA),nitric oxide (NO),inducible nitric oxide synthase (iNOS) and catalase (CAT) were determined by using enzyme assay kits.Hematoxylin-eosin (HE) staining was used to observe morphological changes in brain tissues.Results The survival rate of the sepsis group (30％ at 7th day) was significantly lower than that of the control group (100％ at 7th day)and sham operation group(100％ at 7th day).The survival rate of the sepsis therapy group (45％ at 7th day)was significantly higher than that of the sepsis group(P ＜ O.05).The neurological reflex assessment began to decrease at 6 h in sepsis group and reached the lowest at 24 h (6.00 ± 0.53).The sepsis therapy group (7.62 ± 0.52) was significantly higher (P ＜ 0.05) than the sepsis group and began to recover at 72 h (8.63 ±0.52).ROS,SOD,MDA,NO and iNOS in the sepsis group and the sepsis therapy group reached a peak at 24 h,which decreased at 72 h.The value in sepsis therapy group was significantly decreased than that in the sepsis group,and the difference was statistically significant(P ＜0.05).CAT changed in the opposition.The SOD/CAT in sepsis group was the highest 24 h after the operation,while the ratio in sepsis therapy group was significantly improved.SOD/CAT and MDA were positively correlated(r =0.968,P ＜ 0.05).HE staining showed significant damage to the brain tissue structure in the sepsis group,however some improvement was observed in the sepsis therapy group.Conclusion Vitamin C can significantly improve the survival rate and encephalopathy prognosis in the cecal ligation and puncture SD rat models.The mechanism may be related to the reduction of oxidative stress.