Lipopolysaccharide (LPS) was used to induce mouse mononuclear macrophages (RAW 264.7 cells) to establish the inflammation model for investigating the effect and mechanism of Mailuoning oral liquid on arteriosclerosis occlusion (ASO) in vitro. RAW264.7 cells viability was measured by MTT assay. NO concentration was determined by Griess. mRNA levels and protein expressions of NFAT5/NLRP3 signaling pathway were detected by Q-PCR and Western blot. The relationship between NFAT5 and NLRP3 was explored by cellular transfection of NFAT5-siRNA combined with Western blot. Nuclear translocation of NFAT5 was detected by immunofluorescence. The results showed that Mailuoning oral liquid decreased the NO release induced by LPS in RAW264.7 cells. The mRNA levels of NFAT5, NLRP3, caspase1, IL-18 and MMP9, the protein expressions of NFAT5, NLRP3, cleaved-caspase1 (p20) and the phosphorylation of NF-κB-P65 were decreased after administration of Mailuoning oral liquid. NFAT5-siRNA significantly reversed the increase in protein expressions of NLRP3 induced by LPS in RAW264.7 cells. Both Mailuoning oral liquid and KRN2 (NFAT5 inhibitor) could inhibit the expressions and nuclear translocation of NFAT5. In conclusion, Mailuoning oral liquid exert significant anti-inflammatory effects in vitro by inhibiting the NFAT5/NLRP3 signaling pathway, and NFAT5 might be involved in regulating the expressions of NLRP3.

ObjectiveTo explore the clinical efficacy and safety of Fuzheng Huaji Longbi decoction in treating benign prostatic hyperplasia （BPH） in the patients with the syndrome of healthy Qi deficiency and blood stasis. MethodA total of 94 BPH patients were randomized into control and observation groups， with 47 patients in each group. The control group was treated with doxazosin mesylate sustained-release tablets， and the observation group with Fuzheng Huaji Longbi decoction on the basis of the therapy in the control group. After eight weeks， the international prostate symptom score （IPSS）， quality of life （QOL） score， residual urine volume （RUV）， maximum urinary flow rate （Q_max）， TCM syndrome score， TCM symptom score， electrocardiogram， and liver and kidney function were determined to evaluate the clinical efficacy and safety of the two groups. ResultAfter 8 weeks of treatment， the total response rate in the control group was 63.64% （28/44）， which was lower than that （84.44%， 38/45） in the observation group （χ²=5.026， P<0.05）. The clinical efficacy in the observation group was higher than that in the control group （Z=-2.17， P=0.030）. The treatment in both groups decreased the IPSS， QOL score， RUV， and TCM syndrome scores and increased the Q_max （P<0.05）. Moreover， the observation group had lower IPSS， QOL score， RUV， and TCM syndrome score （P<0.05） and higher Q_max than the control group after treatment （P<0.05）. The treatment in the observation group decreased all the TCM symptom scores （P<0.05）， while that in the control group only decreased the frequency of urination at night and the scores of dysuria， weak urine stream， and post-urinary drainage （P<0.05）. After treatment， the observation group had lower frequency of urination at night and lower scores of mental fatigue， cold limbs， lower abdominal discomfort， and loose stool than the control group （P<0.05）. No adverse events associated with the administration of Fuzheng Huaji Longbi decoction were observed during the treatment period. ConclusionFuzheng Huaji Longbi decoction is effective in treating BPH in the patients with the syndrome of healthy qi deficiency and blood stasis. It can relieve the clinical symptoms and improve the quality of life， being a safe and reliable choice for clinical application.

ObjectiveTo evaluate the clinical efficacy and safety of traditional Chinese medicine （TCM） suppository combined with Yishen Tongluo Qingkang decoction in the treatment of immune infertility. MethodA total of 116 patients meeting the inclusion criteria of this study were randomly divided into an observation group （58 cases） and a control group （58 cases）. The observation group was treated with TCM suppository combined with Yishen Tongluo Qingkang decoction，and the control group was treated with prednisone acetate tablets. Both groups were treated for 12 weeks and followed up six months after treatment. Semen samples of the patients were collected before and after treatment，and the pregnancy status of their spouses，negative conversion rate of seminal plasma anti-sperm antibody （AsAb），sperm concentration，motility，percentage of forward motile sperm，sperm acrosin activity， and incidence of adverse reactions were compared between the two groups. ResultA total of 104 patients completed the study，including 53 cases in the observation group and 51 cases in the control group. Before treatment，the baseline data of the two groups were balanced. After treatment，the total effective rate of the observation group was 92.45%，which was higher than that of the control group （76.47%）（P<0.05），and the negative conversion rate of AsAb in the observation group was higher than that in the control group，but the difference was not statistically significant. After treatment，the sperm motility， percentage of forward motile sperm， and sperm acrosin activity increased in the two groups（P<0.05），and the sperm concentration in the observation group increased （P<0.05）. There was no significant difference in sperm concentration in the control group. After treatment，the sperm concentration，motility，percentage of forward motile sperm， and acrosin activity in the observation group were better than those in the control group （P<0.05）. During the trial，the incidence of adverse reactions in the observation group was lower than that in the control group （P<0.05）. ConclusionTCM suppository combined with Yishen Tongluo Qingkang decoction can significantly increase the negative conversion rate of AsAb and improve the quality of semen in patients with immune infertility.

Objective By employing surgically induced varicocele(VC)in SD rats and an apomorphine(APO)test,we screened rats with erectile dysfunction(ED)after VC and explored method to establish VC and ED models.Methods Sixty rats were randomly divided into Control,Sham,and Model groups with 20 rats in each group.Using the Turner method,we partially ligated the left renal vein to induce left VC three times.APO tests were conducted to screen rats with ED after inducing VC.The numbers of erections,genital grooming,and yawning were observed and recorded.The diameter of bilateral spermatic veins were measured.Both testises and kidneys were weighed.HE staining was used to observe pathological changes of penis and left testis.The success rate of modeling was calculated in the Model group.Results A VC and ED model was successfully established in 15 out of 20 rats in the Model group with a success rate of 75％.After modeling,the diameter of the left spermatic vein in the model group was increased significantly(P＜0.01)and was significantly larger than that before modeling(P＜0.01).The diameter of the right spermatic vein in the Model group was increased(P＜0.05)and higher(P＜0.05)than that before modeling.The weight of the left testis in the Model group was significantly decreased(P＜0.01)compared with that of the right testis.No significant difference in the bilateral kidney weights were observed between or within groups(P＜0.05).In the Model group,the numbers of erections,yawning,and genital grooming decreased significantly(P＜0.01)with the time of modeling.Pathological changes of the left testis and penis were significant in the Model group.Conclusions The Turner method increases the diameter of the spermatic vein in rats,causing testis injury and weight loss,and APO tests can be used to screen rats with ED after VC induction.The combination of the two method is suitable to establish an animal model of VC with an ED status similar to humans.

Acute kidney injury (AKI) is an important factor for the occurrence and development of CKD. The protective effect of dihydroartemisinin on AKI and and reported mechanism have not been reported. In this study, we used two animal models including ischemia-reperfusion and UUO, as well as a high-glucose-stimulated HK-2 cell model, to evaluate the protective effect of dihydroartemisinin on premature senescence of renal tubular epithelial cells in vitro and in vivo. We demonstrated that dihydroartemisinin improved renal aging and renal injury by activating autophagy. In addition, we found that co-treatment with chloroquine, an autophagy inhibitor, abolished the anti-renal aging effect of dihydroartemisinin in vitro. These findings suggested that activation of autophagy/elimination of senescent cell might be a useful strategy to prevent AKI/UUO induced renal tubular senescence and fibrosis.
Animals ; Kidney ; Acute Kidney Injury/chemically induced* ; Ischemia ; Reperfusion Injury/drug therapy* ; Autophagy ; Reperfusion

Abstract OBJECTIVE To explore the material basis and mechanism of Crataegi Fructus in improving metabolic hypertension(MH) by using network pharmacology and molecular docking technique.METHODS The components of Crataegi Fructus were collected by HERB, ETCM database and literature survey; screening all ingredients of Crataegi Fructus to improve MH targets through databases such as SwissTargetPrediction and GeneCards; build "active ingredient-target-disease" network of Crataegi Fructus with Cytoscape software; DAVID was used to analyze GO enrichment and KEGG pathway. The core components and core targets were verified by molecular docking with Autodock software. RESULTS The total of 89 active components were screened from Crataegi Fructus and acted on 84 targets. Among them, the core active components of Crataegi Fructus to improve MH were maslinic acid, fomefficinic acid B, linolenic acid, linoleic acid, methyl-n-nonylketone, apigenin, ursolic acid, etc. The core targets were CYP19A1, PPARA, ESR1, PTGS2, PPARG, NR3C1, MMP9, TNF, etc. The mechanism of action mainly involved multiple signaling pathways such as inflammation, glycolipid metabolism, and vascular endothelial function. Molecular docking showed that the core active ingredients of Crataegi Fructus had high affinity with core targets. CONCLUSION Crataegi Fructus may regulate multiple signaling pathways such as TNF, IL-17, AGE-RAGE, HIF-1, cGMP-PKG through multi-component regulation, thereby inhibiting inflammatory response, improving glucose and lipid metabolism abnormalities, and improving vascular endothelial function, so as to comprehensively exert the role of improving MH in various aspects.

Objective:To investigate the incidence and determinants of vaccine hesitancy towards national immunization program in China and understand the current status of parents' hesitancy to different vaccines used in national immunization program.Methods:A cross-sectional survey was conducted in Beijing, Sichuan and Gansu. The methods of proportional probability sampling and convenience sampling were used to select the eligible study subjects for questionnaire surveys.Results:A total of 3 592 parents were enrolled in the study, in whom 38.22% fully accepted all the vaccines, 59.35% agreed to let their children to receive all the vaccines but showed slight concern, and 2.42% had hesitancy to the vaccines. The vaccine with the most hesitancy was polio vaccine (0.89%), followed by diphtheria pertussis tetanus vaccine (0.70%) and hepatitis A vaccine (0.64%). The dominant reason for vaccine hesitancy was the risk-benefit perception of vaccination (31.03%), followed by the low awareness of the parents (21.84%) and the inconvenience caused by distance and time (21.84%).Conclusions:The incidence of vaccine hesitancy towards national immunization program was low in parents in China, but over 50% of the parents showed concern to the vaccines. It is essential to improve the service quality of national immunization program and strengthen the health education about the vaccination to reduce the incidence of vaccine hesitancy in parents.

Objective To assess the association of polymorphism at rs12645561 locus of Nei endonuclease Ⅷ-like 3 ( NEIL3) gene with the severity of coronary atherosclerosis in Han population of Hubei and Henan regions in central China. Methods In 947 cases undergoing coronary angiography, the severity of coronary atherosclerosis was calculated by lesion vessel number scores and Gensini scores.The genotypes of rs12645561 were analyzed by high resolution melting curve. The plasma NEIL3 levels were measured by en-zyme-linked immunosorbent assay. Results The minor allele T of rs12645561 was significantly associated with the risk of coronary ar-tery disease (χ2=12.165, P<0.05) including increased lesion vessel scores and Gensini scores (χ2=14.745 and 15.615,P<0.05). The variant risk genotypes ( CC+CT) of rs12645561, body mass index of more than 25 kg/m2 , hypelipidemia and smoking habit were all the independent risk factors for higher Gensini scores ( OR=1.50, 1.54, 2.01 and 1.42, respectively, P<0.05) . There were signifi-cantly inverse correlations of plasma NEIL3 levels with the distribution of rs12645561, lesion vessel number scores and Gensini scores ( P<0.05) . Conclusion rs12645561 may correlate with the severity of coronary atherosclerosis, and contribute to the development of coronary atherosclerosis of Han population of Henan and Hubei regions. rs12645561 may also affect the levels of NEIL3 protein.

ObjectiveToinvestigatetheprotectiveeffectandmechanismofscutelarincombinedwith paeoniflorin after permanent cerebral ischemia in rats. Methods Forty-eight adult male SD rats w ere randomly divided into four groups: sham-operation, cerebral ischemia, scutelarin+ paeoniflorin, and cyclopamine (n=12 in each group). A model of permanent middle cerebral artery occlusion w as induced by suture method. The intraperitoneal injection of cyclopamine 6 mg/kg, a specific inhibitor of sonic hedgehog (SHH) pathw ay, at 15 min before ischemia in the cyclopamine group, w hile other groups w ere intraperitoneal y injected an equal volume of saline. At 0 hour and 3 hours after ischemia, the scutel arin+paeoniflorin group and cyclopamine group w ere intraperitoneal y injected scutel arin ( 20 mg/kg ) and paeoniflorin (30 mg/kg), while other groups were intraperitonealy injected an equal volume of saline. Neurological deficit scores w ere performed at 24 hours after ischemia, and then the rats w ere decapitated. The cerebral infarct volume w as measured by using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Real-time fluorescent quantitative polymerase chain reaction and Western blotting w ere used respectively to detect the expression levels of SHH, Patched-1, Gli-1 mRNAs and proteins in the ischemic cortex. Results The neurological deficit scores in the cerebral ischemia group, scutel arin+paeoniflorin group, and cyclopamine group w ere 3.33 ±0.52, 1.50 ±0.55, and 3.67 ±0.52, respectively. The neurological deficit score in the scutel arin+paeoniflorin group w as significantly low er than that in the cerebral ischemia group ( P<0.05), and the neurological deficit score in the cyclopamine group w as significantly higher than that in the scutelarin+paeoniflorin group ( P<0.05). The infarct volume percentage in the cerebral ischemia group, scutelarin+paeoniflorin group, and cyclopamine group were 31.77%±1.19%, 22.94%±2.65%, and 35.53%±0.20%, respectively. The infarct volume in the scutel arin+paeoniflorin group w as significantly less than that in the cerebral ischemia group ( P<0.05), and the infarct volume in the cyclopamine group was significantly larger than that of the scutelarin+paeoniflorin group (P<0.05). The expression levels of SHH, Patched-1, Gli-1 mRNAs and proteins in the cerebral ischemia group, scutelarin+paeoniflorin group, and cyclopamine group w ere significantly higher than those in the sham -operation group (al P<0.05). The expression levels of SHH, Patched-1, Gli-1 mRNAs and proteins in the scutelarin+paeoniflorin group were significantly higher than those in the in the cerebral ischemia group (al P<0.05), and the expression levels of Gli-1 mRNA and protein in the cyclopamine group were significantly lower than those in the scutelarin+paeoniflorin group ( al P<0.05 ). Conclusions The scutel arin combined w ith paeoniflorin has certain protective effect on focal cerebral ischemia injury in rats. Its mechanism is associated w ith the activation of SHH signaling pathw ay.

Objective To analyze the characteristics of fat infiltration into the muscles of patients with Becker and Duchenne muscular dystrophy (DMD) so as to provide a guide for rehabilitation therapy.Methods Twenty-three children with Becker muscular dystrophy (BMD) and 47 with DMD who had never been treated with glucocorticoids were enrolled.MRI was performed on both of their thigh muscles.T1 weighted images were used to assess the fat infiltration of their thigh muscles using a 0-5 modified version of Mercuri's scale.The progression of fatty infiltration of the thigh muscles in BMD was analyzed using descriptive statistics.The differences in fat infiltration between BMD and DMD were analyzed using rank sum tests.Results In patients with BMD the adductor magnus most often showed severe fat infiltration,followed by the biceps femoris,quadriceps,semimembranosus and semitendinosus,while the sartorius,gracilis and adductor longus had the lowest percentages of severe fat infiltration.Among the BMD patients the adductor magnus,biceps femoris and quadriceps showed moderate to severe involvement at the age of 8 to 9.The semimembranosus and semitendinosus showed moderate to severe involvement at the age of 10 to 11,and the sartorius,gracilis and adductor longus showed mild to moderate involvement after 15 years of age.Among the age groups of 8,9,10 and 11 years old,the median total fat infiltration scores were 10,22,28 and 25 respectively among the BMD patients,and 29,34,34 and 30 respectively among the DMD patients.At age 8 significant differences between the BMD and DMD patients were observed in the infiltration scores of the adductor magnus,biceps femoris,vastus lateralis,rectus femoris,vastus medialis,vastus intermedius and in the total scores.At age 9 significant differences persisted in the scores of the adductor magnus,rectus femoris,vastus medialis,vastus intermedius and the total scores.Conclusions The muscle MRIs showed significant differences in the degree of fatty infiltration between BMD and DMD patients.These findings may be useful when designing therapeutic regimens and rehabilitation programs for patients with BMD and DMD.