The level of serum fibroblast growth factor-21 (FGF-21) in patients with type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease (NAFLD) was determined by ELISA.The results showed that serum FGF-21 level in these patients was higher than that in type 2 diabetic patients [(266.55 ± 21.24 vs 220.32 ± 22.68) ng/L,P＜ 0.01].Serum FGF-21 levels in both groups were significantly higher than that in normal control group [(173.52 ± 16.18) ng/L,P＜0.01].Serum FGF-21 level was positively correlated with waist circumference,blood glucose,and triglyceride.FGF-21 may contribute to the development of NAFLD in the patients with type 2 diabetes mellitus.

Objective:To investigate the anti-inflammation effects by activation of the cholinergic anti-inflammatory pathway and its mechanisms in non-alcoholic steatohepatitis (NASH)model mice.Me-thods:6-week-old male C57BL/6J (B6)mice were randomly divided into four groups:the first group was normal mice,injected with saline;the second group was normal mice,injected with nicotine;the third group was NASH model mice,injected with saline;the fourth group was NASH model mice,injec-ted with nicotine.The experimental mice were fed with either standard chow (SC)or high-fat and high-fructose (HFHF)for 17 weeks to generate an NASH model mice.The mice received injection once daily for 3 weeks [nicotine dose,400 μg/kg].Then,their pathological characteristics and function of the liver were assessed.The expressions of interleukin-6 (IL-6)and tumor necrosis factor-α(TNF-α)in se-rum were analyzed by enzyme linked immunosorbent assay (ELISA).The expressions of alpha 7 nicotinic acetylcholine receptors (α7nAChR),Toll-like receptors-4 (TLR-4)and nuclear factor κB of phosphory-lation (p-NF-κB)in Kupffer cells were determined by Western blot and immunofluorescence assays.Re-sults:We successfully generated NASH model mice by imitating the high-fat and high-fructose dietary style of NASH patients.The results of our investigation demonstrated that nicotine could reduce signifi-cantly the levels of IL-6,and TNF-αin serum (P <0.05).The expression of p-NF-κB protein in the group which was NASH model mice injected with nicotine declined significantly as compared with the group which was NASH model mice injected with saline (P <0.05).And the expression of α7nAChR protein elevated significantly conversely (P <0.05 ).Conclusion:Activation of the cholinergic anti-inflammatory pathway could inhibit the release of inflammatory factors as TNF-αand IL-6 in NASH model mice,and the mechanism for the inhibition of inflammatory was mediated by NF-κB pathway.

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

Objective To investigate the effects and possible mechanisms of nicorandil on lung injury of the collapsed lung in one-lung ventilation.Methods Twenty-four clean Japanese big-ear rabbits were randomly divided into sham group (group S) (two-lung ventilation + thoracotomy),negative control group (group C) (one-lung ventilation + thoracotomy + saline),nicorandil group (group N) (one-lung ventilation+thoracotomy+nicorandil) and antagonist group (group J) (onelung ventilation+ thoracotomy+ nicorandil+ glyburide) equally.Mechanical ventilation was implemented through self-made double-lumen endotracheal tube after intravenous induction through ear marginal vein.Intravenous maintenance medicine was infused by trace injection pump after anesthesia induction.Thoracic surgery was simulated through one-lung or two-lung ventilation determined by auscultation,bubble test and direct observation.Then wet and dry weight ratio (W/D) and content of MDA were measured after non-ventilatory lung was processed and preserved.The expression of Akt,p-Akt and NF-κB protein in non-ventilatory lung tissue were detected by Western-blot in all groups.Results In respects of W/D and content of MDA,the other three groups had significant differences compared with group S (P ＜ 0.05).It was significantly lower in group N than in group C (P ＜0.05),and it was significantly higher in group J than in group N (P＜0.05).The expressions of pAkt protein and p-Akt/Akt in group N were significantly higher than those in group S and group C (P＜0.05).Those of group J were significantly lower than group N (P＜0.05).Compared with group S,the expression of NF-κB protein in group C was significantly higher (P＜0.05).That of group N was significantly lower than that of group C (P＜0.05).But that in group J was higher than that in group N (P ＜ 0.05).Conclusion Nicorandil has a protective effect on the collapse and inflation of non-ventilatory lung in rabbits under one-lung ventilation,acting on mitoKATP through PI3K/Akt,and down-regulating NF-κB to reduce IR-induced lung injury.

Objective To investigate the effects of nicorandil on hypoxia-inducible factor (HIF)-1α mRNA and protein in lung tissue of one-lung ventilation.Methods Twenty-four clean New Zealand white rabbits were randomly divided into sham group (group S) (two-lung ventilation+thoracotomy),negative control group (group C) (one-lung ventilation + thoracotomy + saline),nicorandil group (group N) (one-lung ventilation+ thoracotomy+ nicorandil) and antagonist group (group J) (one-lung ventilation + thoracotomy + nicorandil + glibenclanide) equally.The implementation of mechanical ventilation depended on self-made double-lumen endotracheal tube after intravenous induction through ear marginal vein.Intravenous maintenance medicine was infused by trace injection pump after anesthesia induction.The implementation of thoracic surgery was simulated through one-lung and two-lung ventilation by auscultation,bubble test and direct observation.Group S was given anaesthesia only,no one-lung ventilation group S,the other three groups had single lung ventilation,and the drug was injected before the operation.Group N was infused nicorandil 100 ptg· kg-1 · h-1 before the implementation of single lung ventilation for 1 h.Group C was injected with the same amount of normal saline.Group J was intravenous infusion of glibenclamide 75 μg· kg-1 · h-1 and nieorandil 100μg · kg-1 · h-1 the implementation of single lung ventilation for 1 h.Then wet and dry weight ratio(W/D) and superoxide dismutase (SOD) activity were measured after non-ventilatory lung was processed and preserved.The expression of HIF-1α protein of non ventilatory lung tissue was detected by Western-blot in the four groups.The transcription of HIF-1α mRNA was detected by real-time quantitative real-time PCR (qRT-PCR) in all groups.Results W/D in groups C and J were significantly higher compared with that of groups S and N (P＜0.05).The activity of SOD in groups C and J was significantly lower compared with groups S and N (P＜0.05).The expression of HIF1α protein and transcription of HIF-1α mRNA in groups C,N and J were significantly higher than those in group S,and that of group N was significantly higher than those of groups C and J (P＜0.05).Conclsion Nicorandil has a protective effect on the collapse and inflation of non-ventilatory lung in rabbit with one-lung ventilation,reducing oxidative stress by SOD,acting on mito KATP and coming into play by up-regulation of HIF-1α.

Objective:To investigate the efficacy of therapeutic plasma exchange with continuous renal replacement therapy in patients with early septic shock.Methods:A total of 55 patients with septic shock admitted to ICU of Mindong Hospital Affiliated to Fujian Medical University from December 2017 to December 2020 were retrospectively analyzed. The patients were divided into the therapeutic plasma exchange group ( n=29) and standard-therapy group ( n=26) according to whether plasma exchange combined with hemofiltration was used. Patients in both groups were treated according to the 2016 Surviving Sepsis Campaign guidelines. No hemofiltration or/and plasma exchange therapy was performed in the standard-therapy group. In the therapeutic plasma exchange group, hemofiltration was performed immediately after plasma exchange within 24 h. The inflammatory indexes, hemodynamic indexes, organ function scores and 28-day mortality were monitored before and 24 h after treatment. χ2 test was used for counting data, t-test was used for measurement data, and Kaplan-Meier curve was used to evaluate 28-day survival status. Results:(1) There were no differences in sex, age, underlying diseases, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score before treatment between the two groups. (2) There were no significant differences in PCT, CRP and IL-6 between the two groups at enrollment. After treatment, PCT, CRP and IL-6 in the therapeutic plasma exchange group were significantly lower than those in the standard-therapy group [PCT (ng/mL): (50.07±14.54) vs. (57.93±13.42), P=0.043; CRP (mg/L): (85.71±46.05) vs. (115.10±44.42), P=0.042; IL-6 (pg/mL): (5 957.45±2 344.48) vs. (7 522.94±3 218.94), P=0.043], but there was no significant difference in WBC between the two groups. (3) There were no significant differences in norepinephrine dose, mean arterial pressure, heart rate, systemic vascular resistance index (SVRI), extravascular lung water index (EVLWI) and Lactate level between the two groups. After treatment, the norepinephrine dose, lactate level and EVLWI in the therapeutic plasma exchange group decreased significantly, while SVRI increased significantly {norepinephrine dose [μg/(kg·min): (0.76±0.39) vs. (0.54±0.39), P=0.044; lactate (μmmol/ L): (7.74±4.22) vs. (4.51±1.62), P<0.001; EVLWI (mL/kg): (10.04±2.77) vs. (8.23±2.23), P=0.008; SVRI (dyn·s/cm 2): (1 103.14±364.94) vs. (1 403.31±264.46), P=0.001}. Compared with the standard-therapy, the 24-h intravenous infusion volume was significantly decreased [(3 852.07±686.43) mL vs. (4 474.81±572.71) mL, P=0.001]. (4) There were no significant differences in APACHEⅡscore and SOFA score between the two groups at enrollment. After treatment, the APACHEⅡscore and SOFA score of the therapeutic plasma exchange group were significantly lower than those of the standard-therapy group [APACHEⅡscore: (14.07±4.30) vs. (19.23±5.44), P<0.001; SOFA score: (9.93±1.16) vs. (11.69±1.81), P<0.001)]. There were no significant differences in ICU mortality and 28-day mortality between the two groups ( P>0.05). Conclusions:Therapeutic plasma exchange with continuous renal replacement therapy can reduce the inflammatory response and improve hemodynamics in patients with septic shock. However, 24 h treatment did not improve the mortality of patients.

Objective:To investigate the quality of life in children with McCune-Albright syndrome (MAS) and its influencing factors, so as to provide scientific basis for improving their quality of life.Methods:The clinical data of 31 children with MAS diagnosed (MAS group) and followed up in Henan Children′s Hospital from June 2015 to December 2021 were retrospectively analyzed.During the same period, 37 healthy age and sex-matched children at a ratio of 1∶1 were recruited as healthy control group.The children′s Quality of Life Universal Core Scale (PedsQL?4.0) was used for the investigation and comparative analysis.Statistical analysis was performed using the independent sample t test, Chi- square test and multiple regression analysis. Results:Compared with the healthy control group, the physiological function ( t=2.092, P<0.05), emotional function ( t=2.373, P<0.05) and total score ( t=2.360, P<0.05) of MAS group significantly decreased.Multiple regression analysis showed that physiological function was negatively correlated with the annual number of vaginal bleeding ( t=-2.367, P<0.05) and the age of first fracture ( t=-2.606, P<0.05). Social function was negatively correlated with the number of fractures ( t=-2.481, P<0.05). Conclusions:The overall quality of life of MAS children is low, especially the quality of physiological function and emotional function.The annual number of vaginal bleedings, the age of the first fracture and the number of fractures are influencing factors for the reduction of the quality of life of children with MAS.

ObjectiveTo investigate the effect and possible mechanism of Sishenwan in ameliorating visceral sensitivity in the rat model of diarrhea-predominant irritable bowel syndrome （IBS-D） due to spleen-kidney Yang deficiency. MethodForty male SPF-grade rats were randomly assigned into five groups： blank control， model， low- （3.51 g·kg^-1） and high-dose （7.02 g·kg^-1） Sishenwan， and Peifikang （0.54 g·kg^-1） groups. Except the blank control group， the other groups underwent maternal separation stress and Sennae Folium decoction gavage for the modeling of IBS-D due to spleen-kidney Yang deficiency. After corresponding drug interventions， the general conditions of the rats were observed， and the number of defecation pellets within 6 h and the minimum threshold of abdominal withdrawal reflex （AWR） were measured. Enzyme-linked immunosorbent assay （ELISA） was used to measure the serum levels of tumor necrosis factor （TNF）-α， gastrin （GAS）， corticosterone （CORT）， and adrenocorticotropic hormone （ACTH）. Hematoxylin-eosin （HE） staining was employed to observe pathological changes in the colon tissue. Toluidine blue staining was used to assess mast cell degranulation in the colon tissue. Western blot was performed to determine the protein levels of p38 mitogen-activated protein kinase （MAPK）， c-Jun N-terminal kinase （JNK）， transient receptor potential vanilloid 1 （TRPV1）， and protease-activated receptor 2 （PAR2） in the colon tissue. Immunohistochemistry was employed to measure the protein level of TRPV1 in the colon tissue， and immunofluorescence was used to detect the positive expression of substance P （SP） and calcitonin gene-related peptide （CGRP） in the colon tissue. ResultCompared with the blank control group， the model group showed increased number of defecation pellets within 6 h （P<0.01）， decreased minimum threshold of AWR （P<0.01）， elevated serum TNF-α level （P<0.01）， lowered levels of GAS， CORT， and ACTH （P<0.05， P<0.01）， increased mast cell degranulation rate （P<0.01）， increased positive expression of TRPV1， SP， and CGRP （P<0.05， P<0.01）， and upregulated protein levels of p38 MAPK， JNK， TRPV1， and PAR2 （P<0.01）. Compared with the model group， the high-dose Sishenwan group showed increased minimum threshold of AWR （P<0.01）， reduced defecation frequency in both the high-dose Sishenwan and Peifikang groups （P<0.05， P<0.01）， lowered TNF-α level （P<0.05， P<0.01）， elevated levels of GAS， CORT， and ACTH （P<0.05， P<0.01）， decreased mast cell degranulation rate （P<0.01）， reduced positive expression of TRPV1， SP， and CGRP （P<0.05， P<0.01）， and downregulated protein levels of p38 MAPK， JNK， TRPV1， and PAR2 （P<0.05， P<0.01）. ConclusionSishenwan can ameliorate visceral sensitivity in the rat model of diarrhea-predominant irritable bowel syndrome due to spleen-kidney Yang deficiency by regulating the p38 MAPK/JNK/TRPV1 signaling pathway.

ObjectiveTo explore the therapeutic effect and mechanism of intermittent theta burst transcranial magnetic stimulation (iTBS) on non-fluent aphasia after stroke. MethodsFrom August, 2021 to August, 2022, 50 patients with non-fluent aphasia after stroke in the First People's Hospital of Lianyungang were randomly divided into sham stimulation group (n = 25) and iTBS group (n = 25). Both groups accepted speach training. iTBS group accepted iTBS, and the sham stimulation group received sham iTBS, for four weeks. The serum brain-derived neurotrophic factor (BDNF) was measured, and they were assessed by China Rehabilitation Research Center Standard Aphasia Examination (CRRCAE) and Boston Diagnostic Aphasia Examination before and after treatment. ResultsTwo cases in the sham stimulation group and three cases in iTBS group dropped down. The BDNF level, and listen and understand, repeat, name, read aloud, and total score of CRRCAE improved in two groups after treatment (|t| > 5.012, P < 0.001); and they were better in iTBS group than in the sham stimulation group (|t| > 3.968, P < 0.001). The total effective rate was more in iTBS group than in the sham stimulation group (χ² = 8.835, P < 0.05). ConclusioniTBS can improve speech function in patients with non-fluent aphasia after stroke, which may associate with the promotion of BDNF.