OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment of "biaoben acupoint combination" on cardiomyocyte mitochondrial fission in the rats with myocardial ischemia-reperfusion injury (MIRI) and explore its mechanism. METHODS: Fifty male SD rats were randomly divided into a sham-operation group, a model group, an EA pretreatment group, an EA pretreatment + Compound C group and an EA pretreatment+ML385 group, 10 rats in each group. In the EA pretreatment, the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, EA was delivered at bilateral "Neiguan" (PC6), "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, with continuous wave and 2 Hz of frequency, 1 mA of current, once daily for consecutive 7 days. On day 8, in the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, 30 min before model preparation, the intraperitoneal injection with Compound C (0.3 mg/kg) and ML385 (30 mg/kg) was administered respectively. Except in the sham-operation group, the ligation of the left anterior descending coronary artery was performed to prepare MIRI rat model in the rest groups. In the sham-operation group, the thread was not ligated. After modeling, the content of reactive oxygen species (ROS) in the ischemic area was measured by flow cytometry, superoxide dismutase (SOD) was detected using xanthine oxidase method, and malondialdelyde (MDA) was detected using thiobarbituric acid (TBA) chromatometry. The morphology of myocardial tissue in the ischemic area was observed with HE staining, and the mitochondria ultrastructure of cardiomyocytes observed under transmission electron microscopy. Using immunofluorescence analysis, the positive expression of mitochondrial fission factor (MFF), mitochondrial fission 1 protein antibody (Fis1) and dynamin-related protein 1 (Drp1) was detected; and with immunohistochemical method used, the protein expression of adenosine monophosphate-activated protein kinase (AMPK), nuclear factor E2-associated factor2 (Nrf2) and Drp1 in the ischemic area was detected. RESULTS: Compared with the sham-operation group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 increased in the model group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 decreased (P<0.01), and the protein expression of Drp1 elevated (P<0.01). Compared with the model group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01), and the protein expression of Drp1 declined (P<0.01); and in the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the positive expression of MFF, Fis1 and Drp1, and the protein expression of Drp1 were all reduced (P<0.01). When compared with the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01, P<0.05), and the protein expression of Drp1 decreased (P<0.05). In comparison with the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the cardiac muscle fiber rupture, cell swelling and mitochondrial disorders were obviously alleviated in the EA pretreatment group. The morphological changes were similar among the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group. CONCLUSION Electroacupuncture pretreatment of "biaoben acupoint combination" attenuates myocardial injury in MIRI rats, probably through promoting the phosphorylation of AMPK and Nrf2, inhibiting the excessive mitochondrial fission induced by Drp1, and reducing mitochondrial dysfunction caused by mitochondrial fragmentation and vacuolation.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Myocardial Reperfusion Injury/physiopathology* ; Myocytes, Cardiac/cytology* ; Rats ; Acupuncture Points ; Mitochondrial Dynamics ; Humans ; Reactive Oxygen Species/metabolism* ; NF-E2-Related Factor 2/genetics* ; Superoxide Dismutase/metabolism*

Objective To explore the protective effects of pretreatment with the Mongolian medicine Jiruhen Gurigumu-7(JG-7)and Guangzao Sanwei Tang(GZ-3)on myocardial ischemia-reperfusion injury(MIRI)in mice.Methods 60 male C57BL/6J mice were randomly divided into sham operation(Sham)group,model(Model)group,compound danshen drip pill(CDDP)positive control group,JG-7 group,GZ-3 group,and 12 mice in each group to establish the MIRI model,and the H9C2 cells were randomly divided into Control(normoxic)group,H/R(hypoxia 6 h reoxygenation 14 h)group,H/R+JG-7 group,H/R+GZ-3 group.The mice in each group were tested for cardiac function indexes after 30 min of ischemia,24 h and 7 d of reperfusion,TTC staining to detect infarct area after 24 h of MIRI,HE staining to detect myocardial tissue structure and cellular morphology after 24 h of MIRI,TUNEL apoptosis kit to detect apoptosis of myocardial cells after 24 h of MIRI,Masson staining to detect myocardial fibrosis after 7 d of MIRI.Blood was taken from the abdominal aorta,serum was separated,and the indexes after oxidative stress of MIRI were detected in each group of mice,and the survival rate of H9C2 cells after H/R was detected in each group by CCK-8 method.Results The results of TTC showed that JG-7 and GZ-3 reduced the infarct area after 24 h of MIRI in mice.ELISA and kit assays proved that JG-7 and GZ-3 reduced creatine phosphokinase isoenzyme(Creatinekinase-MB,CK-MB),Lactic dehydrogenase(LDH),malondialdehyde(MDA)levels,and increased superoxide dismutase(SOD)levels.HE staining showed that JG-7 and GZ-3 improved myocardial pathology after MIRI 24 h.The results of TUNEL apoptosis assay showed that JG-7 and GZ-3 improved apoptosis in myocardial tissues 24 h after MIRI.Masson staining results showed that JG-7 and GZ-3 could reduce the area of myocardial tissue fibrosis after MIRI 7 d.CCK-8 assay results showed that JG-7 and GZ-3 could improve the cell survival rate after H/R in H9C2 cells.Conclusion Pre-treatment with Mongolian medicine Jiruhen Gurigumu-7 and Guangzao Sanwei Tang can reduce the damage caused after ischemia-reperfusion(I/R),decrease the area of myocardial infarction and fibrosis after I/R in mice,and protect the heart.

Objective To explore the protective effects of pretreatment with the Mongolian medicine Jiruhen Gurigumu-7(JG-7)and Guangzao Sanwei Tang(GZ-3)on myocardial ischemia-reperfusion injury(MIRI)in mice.Methods 60 male C57BL/6J mice were randomly divided into sham operation(Sham)group,model(Model)group,compound danshen drip pill(CDDP)positive control group,JG-7 group,GZ-3 group,and 12 mice in each group to establish the MIRI model,and the H9C2 cells were randomly divided into Control(normoxic)group,H/R(hypoxia 6 h reoxygenation 14 h)group,H/R+JG-7 group,H/R+GZ-3 group.The mice in each group were tested for cardiac function indexes after 30 min of ischemia,24 h and 7 d of reperfusion,TTC staining to detect infarct area after 24 h of MIRI,HE staining to detect myocardial tissue structure and cellular morphology after 24 h of MIRI,TUNEL apoptosis kit to detect apoptosis of myocardial cells after 24 h of MIRI,Masson staining to detect myocardial fibrosis after 7 d of MIRI.Blood was taken from the abdominal aorta,serum was separated,and the indexes after oxidative stress of MIRI were detected in each group of mice,and the survival rate of H9C2 cells after H/R was detected in each group by CCK-8 method.Results The results of TTC showed that JG-7 and GZ-3 reduced the infarct area after 24 h of MIRI in mice.ELISA and kit assays proved that JG-7 and GZ-3 reduced creatine phosphokinase isoenzyme(Creatinekinase-MB,CK-MB),Lactic dehydrogenase(LDH),malondialdehyde(MDA)levels,and increased superoxide dismutase(SOD)levels.HE staining showed that JG-7 and GZ-3 improved myocardial pathology after MIRI 24 h.The results of TUNEL apoptosis assay showed that JG-7 and GZ-3 improved apoptosis in myocardial tissues 24 h after MIRI.Masson staining results showed that JG-7 and GZ-3 could reduce the area of myocardial tissue fibrosis after MIRI 7 d.CCK-8 assay results showed that JG-7 and GZ-3 could improve the cell survival rate after H/R in H9C2 cells.Conclusion Pre-treatment with Mongolian medicine Jiruhen Gurigumu-7 and Guangzao Sanwei Tang can reduce the damage caused after ischemia-reperfusion(I/R),decrease the area of myocardial infarction and fibrosis after I/R in mice,and protect the heart.

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg^-1·d^-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg^-1·d^-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.

Objective:To investigate whether the existence of thoracic artery blood supply in peripheral pulmonary lesions is the key factor affecting the accuracy of contrast-enhanced ultrasound in differentiating benign and malignant lesions.Methods:From June 2020 to December 2021, a total of 170 patients with peripheral pulmonary lesions were consecutively enrolled in Xi′an Chest Hospital, and all patients underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS). Taking ΔAT(lesion-lung arrival time difference ) of 2.5 seconds as the cut-off point for differentiating benign and malignant lesions(ΔAT<2.5 s for benign, ΔAT≥2.5 s for malignant), and the final pathological results as the gold standard, these patiens were divided into correct classification group and wrong classification group, and the main influencing factors of wrong classification were analyzed.Results:Compared with the correct classification group, the proportion of thoracic artery blood supply in the wrong classification group was significantly higher ( P<0.001). After adjusting the dendritic venous reflux, thoracic artery blood supply was an independent influencing factor for CEUS misclassification ( OR=3.531, 95% CI=1.805-6.908, P<0.001). In the patients with thoracic artery blood supply, the sensitivity of the absence of dendritic venous reflux in judging malignant peripulmonary lesions was 75.0%, the specificity was 91.3%, and the area under the ROC curve (AUC) was 0.832 (95% CI=0.715-0.915, P<0.001), while the sensitivity, specificity and AUC of CEUS in judging malignant peripulmonary lesions were 68.7%, 67.4% and 0.659 (95% CI=0.528-0.775, P=0.006), and there was significant statistical difference in the AUC between them( P<0.001). In the group of patients without thoracic arterial blood supply, the sensitivity of the absence of dendritic venous reflux in judging malignant peripulmonary lesions was 82.8%, the specificity was 63.3%, and the AUC was 0.730 (95% CI=0.636-0.811, P<0.001), while the sensitivity, specificity and AUC of CEUS in judging malignant peripulmonary lesions were 62.1%, 81.0% and 0.684 (95% CI=0.587-0.770, P=0.003), and there was no significant difference in the AUC between them ( P=0.425). Conclusions:The presence of thoracic artery blood supply significantly decrease the diagnostic efficiency of CEUS in differentiating benign and malignant of peripheral pulmonary lesions. For peripheral pulmonary lesions with thoracic arterial blood supply, the diagnostic efficiency of dendritic venous reflux is better than CEUS, while for lesions without thoracic artery supply, the diagnostic efficiency of dendritic venous reflux is equivalent to CEUS.

Objective To estimate the diagnostic value of cytology, DNA-ICM(DNA-image cytometry),cytology combined with DNA-ICM for pancreatic malignancy,and to explore the cut-off value for DNA-ICM. Methods Patients with suspicious pancreatic malignancy were retrospectively identified. In total,145 EUS-FNA specimens acquired from 140 separate patients were examined by cytology and DNA-ICM. Diagnostic values among cytology, DNA-ICM and the combination of the techniques in detecting pancreatic malignancy were compared. Results Compared with cytology, DNA-ICM had a lower sensitivity (63.0% VS 82.4%)and accuracy(69.7% VS 85.5%). After combining the techniques, the diagnostic value for pancreatic malignancy significantly improved compared with that by cytology(0.941 VS 0.912, P=0.007 0)or DNA-ICM only(0.941 VS 0.815, P<0.000 1). By using the Youden index, the cut-off value for DNA-ICM to detect pancreatic malignancy was one cell with DI(DNA index)≥2.5. Notably,with this standard, the sensitivity and accuracy of DNA-ICM significantly increased to 72.3% and 77.2%, and those of the combined techniques increased to 91.6% and 93.1%, respectively. Conclusion Automated DNA-ICM is an objective and effective method for pancreatic malignancy. Although DNA-ICM has a lower diagnostic value than that of conventional cytology, an improved value was obtained after combining the techniques.

Objective To investigate the clinical outcome of pancreatic pseudocyst(PPC)treated with non-surgical methods,and to compare the efficacy and safety between percutaneous drainage and endoscopic drainage in the management of PPC.Methods From February 2010 to July 2017,clinical data of patients with PPC,who received percutaneous drainage or endoscopic drainage,were retrospectively analyzed.The symptom relief rate,short-term and long-term radiologic remission rate,complication rate, recurrence rate and length of hospital stay were compared between patients treated by ultrasound guided percutaneous drainage(percutaneous group),by endoscopic ultrasonography-guided drainage(EUS group)and by endoscopic retrograde pancreatography guided transpapillary drainage(ERP group).Two independent samples t test,one-way analysis of variance,non-parametric test and Fisher′s exact test were performed for statistical analysis.Results A total of 153 patients were treated and the operation was successfully conducted in 148 patients(96.7%),of whom 39 were in percutaneous group,73 in EUS group and 36 in ERP group.The median follow-up time was 26 weeks(two weeks to 358 weeks).The symptom relief rate,long-term radiographic remission rate,complication rate,recurrence rate and retreatment rate of percutaneous group,EUS group and ERP group were 87.2%(34/39),79.5%(58/73),80.6%(29/36);81.5%(22/27),88.6%(39/44),66.7%(16/24);17.9%(7/39),28.8%(21/73),16.7%(6/36);15.0%(3/20),13.8%(8/58),10.0%(2/20);and 10.3%(4/39),8.2%(6/73),2.8%(1/36),respectively.There was no statistically significant difference among three groups (all P> 0.05).The short-term radiographic remission rate of ERP group was significantly lower than those of percutaneous group and EUS group(46.7%,14/30 vs 77.1%,27/35 and 87.7%,64/73),and the differences were statistically significant(χ2 =6.442 and 19.450,both P<0.01).The median hospital stay of percutaneous group was longer than those of EUS group and ERP group(14.0 days vs 9.0 days and 8.0 days),and the differences were statistically significant(Z= -3.687 and -2.630,both P<0.01).Conclusions The efficacies of percutaneous drainage and EUS-guided drainage are both better than ERP,and they are effective and safe methods especially for the patients with complication of pseudocysts and necrosis debris in pseudocysts.However,the hospitalization time of percutaneous drainage is longer. In addition,percutaneous drainage can be an alternative method after failed endoscopic drainage.

Objective To investigate the expression of parvalbumin (PV)in the cognitive im-pairment mice induced by sevoflurane anesthesia,and to explore whether enriched environment could reverse it.Methods One hundred and forty-four Six-day-old C57BL/6 male mice at postnatal day 6 were randomly divided into the following four groups (n =36):control+standard environment group (group CS),control+enriched environment group(group CE),sevoflurane anesthesia+standard en-vironment group(group SS)and sevoflurane anesthesia+enriched environment group(group SE).An-imals were exposed to 3% sevoflurane plus 30% O 2 or 30% O 2 2 h daily for 3 days from postnatal day 6 (P6)to P8.The exposed pups were randomly allocated to an enriched environment for 2 h daily between P8 and P90 or to a standard environment.Western blotting were used for determining PV ex-pression in the prefrontal cortex and hippocampus at P9,P14,P42,P60 and P90.Cognitive functions were assessed by performing the open field (P41 ),and fear conditioning tests (P42-43 and P89-90, respectively).Results In the open field test,there was no significant difference in the total travel dis-tance and the time spent in the center of the arena among groups.In the contextual fear condition test, compared with group CS,the group SS only exhibited a reduced freezing response in the contextual test at P43,but not at P90.In the cued fear conditioning test,no difference was observed in the freez-ing time among the four groups.The PV expression in the prefrontal cortex and hippocampus of group SS were significantly lower than that of group CS at P9 and P14 (P <0.001),while recovered at P60 and P90.The PV expression in the prefrontal cortex and hippocampus of group SE were significantly higher than that of group SS at P42 (P <0.05).Linear regression analysis demonstrated a positive correlation between the PV expression in the prefrontal cortex and the freezing time induced by envi-ronment (r =0.670 7,P =0.000 1),and there was a positive correlation between the PV expression in hippocampus and the freezing time induced by environment (r = 0.509 6,P = 0.001 9 ). Conclusion The cognitive impairment induced by sevoflurane anesthesia may be associated with the reduction of PV in the prefrontal cortex and hippocampus.Placement of the sevoflurane-exposed mice in an enriched environment prevented the development of these abnormalities.

Objective To observe the effects and toxicity of nimotuzumab combined paclitaxel and carboplatin in the treatment of advanced non small cell lung cancer .Methods 168 cases of patients with advanced non-small cell lung cancer were randomly divided into study group and control group ,with 84 cases in each group .Study group was treated with nimotuzumab 200 mg ,1 times per week ,for 6 weeks ,two weeks later the patients were treated with 200 mg and stopped when the disease progressed ,paclitaxel lipo-some(135 mg/m2 ,iv drip 3 h ,1 days) ,carboplatin(300 mg/m2 ,iv drip ,1 days);the control group was given paclitaxel liposome(135 mg/m2 ,iv drip 3 h ,1 days) ,carboplatin(300 mg/m2 ,iv drip ,1 days) ,21 days for 1 cycle .Results Study group RR 38 .1% (32/84);control group RR 23 .8% (20/84)(P<0 .05) .The main side effects of two groups of were gastrointestinal reaction and bone marrow suppression ,there were 64 cases of above Ⅱ digestive tract reaction in the study group ,accounting for 76 .2% ,52 cases ofⅡ degrees of bone marrow suppression ,accounting for 61 .9% ,52 cases of Ⅱ degrees of alopecia ,accounting for 61 .9% ;54 cases of control group Ⅱ digestive tract reaction ,accounting for 66 .7% ,54 cases of Ⅱ degrees of bone marrow suppression ,accounting for 66 .7% ,and 58 cases of Ⅱ degrees of alopecia ,accounted for 57 .1% (P>0 .05) .In the study group ,progression-free survival period was 7 .8 months ,the median survival period was 14 .2 months;in the control group ,progression-free survival period was 5 .1 months ,the median survival period was 9 .8 months(P<0 .05) .Conclusion nimotuzumab combined paclitaxel and carboplatin in the treatment of advanced non small cell lung cancer has good clinical curative effect ,and would not increase adverse reactions .It is worthy of clinical application .

The basical structure of diketopiperazines is a cyclic dipeptide condensed by two amino acids. Because of the stable framework of the six-member ring structure, and having two hydrogen bond donor and two hydrogen bond receptor, DKPs have become important chemical pharmacophores, with strong biological activities and pharmacological activities in the drug. A series of cyclic compounds were found from marine organisms in recent years, research showed that their functions are not limited on anti-bacterial, cytotoxic activity, and so on, but also playing an important role in regulatory mechanism of quorum sensing as signal molecules, they have become research hot point in ecological chemistry. This paper reviewed the research progress of diketopiperazines found in the marine microbial metabolites, and the future study trends was discussed and outlooked.