Objective:To explore the role and the intervention effect of emodin in intestinal neuropathy in rats with severe acute pancreatitis (SAP) through the nucleotide binding oligomerization domain like receptor protein 3/cysteine containing aspartic acid protease-1 (NLRP3/Caspase-1) pathway.Methods:Forty male healthy SD rats aged 6-8 weeks with a weight of approximately 200g were randomly divided into control group, SAP model group, emodin treatment (EMO) group, and NLRP3 knockdown group. SAP were induced by retrograde injection of sodium deoxycholate into the pancreatic duct of rats and serum amylase of which were detected. The effective NLRP3 knockdown sequence was screened for NLRP3 knockdown animal experiments. Fluorescence quantitative polymerase chain reaction was used to detect the expression of NLRP3, Caspase-1, gasdermin-D (GSDMD), interleukin (IL)-1β, IL-18 and tumor necrosis factor-α(TNF-α) in the small intestine of each group. Immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein (GFAP) in the small intestine of each group.Results:The amylase levels of the control group, SAP group, EMO group, and NLRP3 knockdown group were (277.73±24.92) U/L, (1018.57±282.89) U/L, (625.43±134.40) U/L, and (391.01±27.63) U/L, respectively. The SAP and EMO groups were significantly higher than the control group ( P<0.001), while the EMO and NLRP3 knockdown groups were significantly lower than the SAP group (all P<0.001). Compared with control group, the expression levels of NLRP3, Caspase-1, IL-1β, IL-18, TNF-α and GSDMD in SAP group were increased, with statistical significance (all P<0.001). Compared with SAP group, the NLRP3 knockdown group showed the expressionlevels of the above 6 genes were all decreased, and EMO group showed decreased gene expressing levels of NLRP3, IL-1β, IL-18 and TNF-α, with statistical significance (all P<0.05). The relative expression of GFAP in small intestine of control group, SAP group, EMO group and NLRP3 knockdown group were (1.00±0), (1.66±0.11), (1.13±0.02) and (1.13±0.02), respectively. Among them, the expression of GFAP in SAP group was increased compared with the control group; The expression of GFAP in EMO group and NLRP3 knockdown group was lower than that in model group, and the differences were statistically significant (all P<0.05). Conclusions:Emodin and knocking down NLRP3 can both promote the repair of SAP small intestine injury through the NLRP3/Caspase-1 signaling pathway, and thus play a protective role in the intestine.

Objective To investigate the effects of citicoline sodium and Ureicrin on the expression of miR-17-5p and miR-29b in patients with ischemic stroke.Methods A total of 100 patients with ischemic stroke treated in the hospital were divided into control group and combination group by random table method.From February 2023 to February 2024,they were respectively given Eurecrine monotherapy and citicoline sodium and eurecrine combination therapy.Cerebral blood perfusion indexes,inflammatory factors,oxidative stress,miR-17-5p,miR-29b levels and clinical efficacy were detected in the two groups,and adverse reactions were statistically recorded.Results There was no significant difference in CBF,CBV,hs-CRP,miR-29b,IL-6,TNF-α and miR-17-5p between the two groups before treatment(P>0.05).After treatment,the levels of CBF,CBV,miR-29b,SOD and GSH-Px increased,while the levels of hs-CRP,IL-6,TNF-α,miR-17-5p and MDA decreased,and the levels of CBF,CBV,miR-29b,SOD and GSH-Px in the combined group were higher than those in the control group.The levels of hs-CRP,TNF-α,IL-6,miR-17-5p and MDA were significantly different from those of the control group(P<0.05).The incidence of adverse reactions and the total effective rate of treatment in combination group were higher than those in control group(P<0.05),and the adverse reactions were mild and tolerable.Conclusion The combination of citicoline sodium and Ureicrin can improve the clinical efficacy and prognosis of patients.

ObjectiveTo evaluate the effect of the therapy of dispelling stasis， removing toxin， and promoting urination （modified Linggui Zhugantang combined with Xuebijing injection） on the prognosis of sepsis-induced cardiomyopathy （SICM）. MethodA total of 96 patients were randomly assigned into an observation group and a control group， with 48 patients in each group. The patients in the control group received sepsis bundle， and those in the observation group additionally received the therapy of dispelling stasis， removing toxin， and promoting urination （intravenous drip of Xuebijing injection and oral administration of modified Linggui Zhugantang）. The course of treatment in both groups was 7 days. The disease and prognosis indicators ［28-day mortality， intensive care unit （ICU） length of stay， major adverse cardiac events （MACE）， acute physiology and chronic health evaluation Ⅱ （APACHE Ⅱ）， sequential organ failure assessment （SOFA） score， and mortality in emergency department sepsis （MEDS） score］， cardiac function indicators ［left ventricular ejection fraction （LVEF）， E/A ratio of peak velocity blood flow from left ventricular relaxation in early diastole （the E wave） to peak velocity flow in late diastole caused by atrial contraction （the A wave）， E/e′ ratio of mitral peak velocity of early filling （E） to early diastolic mitral annular velocity （e′）， and afterload-corrected cardiac performance （ACP）］， myocardial injury markers ［high-sensitivity cardiac troponin T （hs-cTnT）， N-terminal pro-brain natriuretic peptide （NT-proBNP）， heart-type fatty acid-binding protein （H-FABP）， and high mobility group box-1 （HMGB-1）］， hemodynamic indicators ［extravascular lung water index （EVLWI）， global end-diastolic volume index （GEDVI）， cardiac index （CI）， and systemic vascular resistance index （SVRI）］， and TCM syndrome scores were assessed and compared between the two groups. ResultThe 28-day mortality and the incidence of MACE in the observation group were slightly lower than those in the control group. The ICU length of stay in the observation group was shorter than that in the control group （P<0.05）. After treatment， APACHE Ⅱ， SOFA， MEDS， syndrome score of stasis-caused internal obstruction， E/e′ ratio， hs-cTnT， NT-proBNP， H-FABP， and HMGB1 decreased compared with those before treatment （P<0.05）， while LVEF， E/A ratio， and ACP increased （P<0.05）. Moreover， the changes were more significant in the observation group （P<0.05）. On days 3， 5， and 7 after treatment， the EVLWI and SVRI in the observation group were lower than those in the control group （P<0.05）， while CI showed an opposite trend （P<0.05）. The observation group had higher GEDVI than the control group on days 3 and 5 after treatment （P<0.05）. ConclusionOn the basis of conventional bundle therapy， modified Linggui Zhugantang combined with Xuebijing injection with the effect of dispelling stasis， removing toxin， and promoting urination can inhibit the generation of myocardial injury markers and improve hemodynamics to shorten the length of ICU stay， mitigate the TCM syndrome， and reduce the risk of death， thereby improving the prognosis of SICM.

Objective:To explore the medication rules of Ye Tianshi in the treatment of constipation from his clinical records. Methods:The drugs in the prescriptions of intestinal obstruction and stool closure were collected in Guide to Clinical Practice with Medical Records. Then the frequency of the prescriptions and the herbs for the treatment,as well as the herbal natures, flavors and meridian belongings were analyzed by Excel, SPSS 22.0 and SPSS Modeler software. Results:The main syndrome types of Ye's treatment of fecal occlusive disease were internal accumulation of dampness and heat, deficiency of kidney Yang, hyperactivity of fire due to yin deficiency, deficiency of liver and kidney Yin, and dysfunction of lung. The medicinal properties were mainly warm, cold and flat, and the tastes were mainly sweet, pungent and bitter, and the meridians mainly belonged to spleen and stomach, lung, liver, kidney and heart. Armeniacae Semen Amarum, Angelica sinensis, Poria, Semen Platycladi, Trichosanthes kirilowii Maxim, Semen Persicae were often used. The commonly used drug pairs, included Armeniacae Amarum Semen and Trichosanthes kirilowii Maxim, Armeniacae Amarum Semen and Radix Curcumae Aromaticae, Angelica sinensis and Semen Platycladi and so on. Conclusions:Ye Tianshi takes spleen and stomach as the center, pays attention to the important position of dampness pathogen in the pathogenic process, and pays attention to lung Qi purging, liver Qi drainage and bladder gasification. It provides new treatment ideas and methods, guiding future doctors.

Objective:To explore the effect and mechanism of interleukin-35 gene modified mesenchyma stem cells(MSC)on ameliorating cardiac allograft rejection and prolonging graft survival of transplanted heart in mice.Methods:In this study, IL-35-MSC secreting IL-35 continuously and steadily were successfully constructed in vitro. Abdominal heterotopic heart transplantation model was established successfully. And they were randomly divided into syngeneic control group; saline control group, MSC treatment group and IL-35-MSC experimental group(n=12 each). Six mice were randomly selected for sacrificing at Day 5 post-operation for detecting the related indicators in each group: Hematoxylin eosin staining was used for pathological examination. Enzyme-linked immunosorbent assay(ELISA)was employed for detecting the concentration of IL-35 in peripheral blood and the proportion of T lymphocyte subsets in spleen was analyzed by flow cytometry(FCM). Then the remaining mice were used for recording the graft survival.Results:The model of abdominal heterotopic heart transplantation in mice was successfully constructed. As compared with saline control group(6.50±0.55 d)and MSC treatment group(12.00±0.89 days), IL-35-MSC significantly alleviated rejection after transplantation and effectively prolonged the survival time of graft(18.50±1.64 days)(n=6, P<0.01). As compared with other groups, percentage of Th17 cells and Th1/Th2 ratio in spleen decreased significantly while the proportion of CD4 + Foxp3 + Treg increased significantly in IL-35-MSC experimental group at Day 5 post-transplantation(n=6, P<0.01). Conclusions:IL-35-MSC may alleviate cardiac allograft rejection and prolong graft survival. And cellular immunotherapy based upon IL-35-MSC may provide a new approach for inducing immune tolerance.

Objective To investigate the action mechanism of IL-35 gene transfection ameliorating cardiac allograft rejection and prolonging allograft survival.Methods pEBI3-L-p35-Fc plasmid was amplified by polymerase chain reaction.In vitro plasmid DNA pEBI3-L-p35-Fc or pSec-L-Fc was,respectively,transfected into HEK293 cells using Lipofectamine 3000.At 48 and 72 h after transfection,IL-35 concentration in culture supernatant of transfected HEK293 cells was detected by ELISA.Balb/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of IL-35,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with IL-35 plasmid (50μg) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.The IL-35 expression in the blood,CD4+ CD25+ Tregs proportion in the blood and spleen,and the survival and the histopathologic changes of the cardiac grafts were also observed.Results In vitro the transfected HEK293 cells expressed IL-35.IL-35 enhanced the proliferation of CD4+ CD25+ Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (16 days) was significantly longer than in control group (7 days) (P＜0.01).As compared with control group,CD4+ CD25+ Tregs proportion was significantly increased (P＜0.01),CD8+ T cells proportion was decreased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited in the experimental group.Conclusion IL-35 could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25+ Tregs and inhibition of proliferation of CD8 + effector T cells.

Objective To evaluate the effects of intrathecal low-dose naloxone,morphine and fentanyl on the expression of motillin (MTL) in the hippocampus of rats with incisional pain.Methods Seventy-two healthy male Sprague-Dawley rats,weighing 180-220 g,aged 6-8 weeks,in which intrathecal catheters were successfully implanted,were randomly divided into 6 groups (n =12 each) using a random number table:normal saline group (NS group),incisional pain group (P group),morphine + fentanyl + incisional pain group (MFP group),and naloxone (0.2,1.0 and 5.0 ng/kg) + morphine + fentanyl groups (MFPN1,MFPN2 and MFPN3 groups).Incisional pain was induced by an incision made into the plantar surface of the right hindpaw.At 20 min before induction of incisional pain,the mixture of morphine 5 μg/kg and fentanyl 0.25 mg/kg was injected intrathecally in group MFP,and the mixture of naloxone 0.2,1.0 and 5.0 ng/kg,morphine and fentanyl were injected intrathecally in MFPN1,MFPN2 and MF-PN3 groups,respectively.Six rats in each group were selected,and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before intrathecal catheterization (T0,baseline),at 24 h before induction of incisional pain (T1),and at 1,3 and 6 h after operation (T2-4).The left 6 rats in each group were selected and sacrificed at 6 h after operation,and the hippocampi,body of the stomach and duodenum were removed for detection of MTL content by enzyme-linked immunosorbent assay.Results Compared with group NS,the MWT was significantly decreased,and the TWL was shortened at T2-4 in P and MFPN3 groups,the MWT was significantly decreased,and the TWL was shortened at T4 in group MFPN1,and the TWL was prolonged at T2 in group MFPN2,the MTL contents in hippocampus and body of the stomach were significantly decreased in P,MFP,MFPN1 and MF-PN3 groups,the MTL contents in duodenum were increased in P and MFPN3 groups,and the MTL contents in duodenum were decreased in MFP and MFPN1 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group MFPN2 (P＞0.05).Conclusion Intrathecal naloxone 1.0 ng/kg combined with morphine and fentanyl can inhibit up-regulation of the expression of MTL in the hippocampus of rats with incisional pain,and then is involved in the maintenance of stable gastrointestinal motility.

Objective To investigate the effect of exogenous hydrogen sulfide (H2S) on neuronal apoptosis during focal cerebral ischemia-reperfusion (I/R) in rats.Methods One hundred eight male Sprague-Dawley rats,aged 6-8 weeks,weighing 250-270 g,were randomly divided into 3 groups (n =36 each) using a random number table:control group (group C),I/R group and H2S group.Focal cerebral ischemia was induced by middle cerebral artery occlusion in anesthetized rats.A nylon thread with a rounded tip was inserted into the left internal carotid artery and advanced intracranially to block blood flow into the middle cerebral artery.Middle cerebral artery occlusion was maintained for 90 min followed by reperfusion.In group H2S,0.25％ NaSH (a donor of exogenous H2S) 10 mg/kg was injected intraperitoneally at the onset of reperfusion.The equal volume of normal saline was given in C and I/R groups.At 1,3 and 7 days of reperfusion,neurological deficit was scored,and corner test was performed.Brains were removed for determination of myocardial infarct size,Bax-,Bcl-2-and caspase-3-positive cells,and cell apoptosis.The percentage of myocardial infarct size,rate of Bax-,Bcl-2-and caspase-3-positive cells and apoptosis rate were calculated.Results Compared with group C,the neurological deficit score was significantly decreased,and the corner score,percentage of myocardial infarct size,rate of Bax-,Bcl-2-and caspase-3-positive cells and apoptosis rate were increased at each time point of reperfusion in I/R and H2S groups (P＜0.05).Compared with group I/R,the neurological deficit score and Bcl-2-positive cells were significantly increased,and the corner score,percentage of myocardial infarct size,rate of Bax-and caspase-3-positive cells and apoptosis rate were decreased at each time point of reperfusion in group H2S (P＜0.05).Conclusion The mechanism by which exogenous H2S attenuates focal cerebral I/R is related to inhibition of neuronal apoptosis in rats.

Objective:To explore the independent risk factors of death during hospitalization in patients with acute myocardial infarction (AMI) .Methods :Clinical data of 614 cases ,who were diagnosed as AMI during hospitaliza‐tion in our hospital from 2011 to 2013 ,were retrospectively analyzed .According to AMI patients'survival or not during hospitalization ,they were divided into death group (n=62) and survival group (n=552) ,single and multi-variable Logistic regression analysis were used to analyze the relationship among baseline feature factors and thera‐peutic methods of all patients and hospital mortality .Results:The mean age was (66.58 ± 12.87) years and there were 39 males (62.9% ) among the 62 dead patients .Hospital mortality was 10.10% (62/614) .Multi‐variable Lo‐gistic regression analysis screened following factor were independent risk factors related to AMI hospital mortality :age (OR= 3.065 ,95% CI:1.188~ 7.915) ,female (OR= 2.775 ,95% CI :1.200~ 6.419) ,heart rate (OR=2.836 ,95% CI:1.405~ 5.722) ,blood glucose (OR = 1.943 ,95% CI :1.186 ~ 3.184) ,Killip class IV (OR=1.744 ,95% CI:1.211~2.513) and left main or triple -vessel coronary disease (OR= 3.157 ,95% CI :1.244 -8.014) . P < 0.05 ~ < 0.01 .Conclusion : Advanced age ,female ,rapid heart rate ,elevated blood glucose level at hospitalization ,Killip class IV and left main or triple‐vessel coronary disease may be independent risk factors of death during hospitalization in patients with acute myocardial infarction .

OBJECTIVE:To investigate the improving effect of phenylethanoid glycosides (PhGCs) from Tibetan medicine Phlomis younghusbandii on rats with acute high-altitude cerebral edema. METHODS:60 Wistar rats were randomly divided into a normoxia control group (isometric sterile water for injection),a hypoxia model group (isometric sterile water for injection),a dexamethasone group(4 mg/kg),and three groups of PhGCs at high(400 mg/kg),middle(200 mg/kg)and low(50 mg/kg)dos-es,with 10 rats in each group. The rats were given drugs,ig,6 d before the establishment of models. On the 4th day of administra-tion,ig,the rats in all groups except the normoxia blank group were placed in a simulated 8 000 m altitude plateau environment for 72 h hypoxic exposure to establish the rat models of high-altitude cerebral edema. Following HE stain,the pathological changes in rats’brain tissues were observed under the light microscope. Dry-wet proportion method was used to determine the water con-tents in rats’brain. The content of MDA and the activities of SOD and GSH in rats’brain tissues were detected. Enzyme-linked im-munosorbent assay was adopted to determine the contents of IL-1β and TNF-α in rats’serum and brain tissues. RESULTS:Com-pared to the rats in the normoxia control group,those in the hypoxia model group showed obvious brain edema,and thickened lacu-nas around cells and vessels and inflammatory cell infiltration, higher water contents and MDA and weaker activities of SOD and GSH in brain,and higher contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). Compared to the rats in the hypoxia model group,those in the groups of PhGCs at high,middleand low dosages demonstrated less inflammatory cell infiltration and lower water contents in brain tissues,in which the groups of PhGCs at high and middle dosages demonstrated lower content of MDA and stronger activities of SOD and GSH in brain tissues, and lower contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). CONCLUSIONS:PhGCs can obviously alleviate the acute cerebral injury in rats which is caused by acute hypoxia and has im-provement effect to some degree on the rats with acute high-altitude cerebral edema.