Objective Network pharmacology and molecular docking techniques were used to predict the potential mechanisms by which the active components of Piper nigrum(PN)regulate depressive-like behaviors in chronic restraint stress(CRS)mice.Methods The major chemical components and targets of PN were screened using the Traditional Chinese Medicine Systems Pharmacology database.Targets related to ferroptosis and depression were obtained from the Online Mendelian Inheritance in Man,GeneCards,and FerrDB databases.The intersecting targets were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analyses,and molecular docking was performed to validate the binding capacities between the core targets and their corresponding active components.Finally,we established a CRS mouse model.Mice were treated with PN 75,150,and 300 mg/kg for 4 weeks,followed by behavioral assessments and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)to verify the expression of core genes.Results Nine active components were screened from PN,corresponding to 27 targets,and 8377 targets related to depression and 547 targets associated with ferroptosis were screened from the databases.The intersection of these three sets resulted in 25 target genes.KEGG enrichment analysis revealed that these core targets were predominantly enriched in signaling pathways,including cholinergic synapses,serotonergic synapses,and neuroactive ligand-receptor interactions.Molecular docking result showed that the main active components of PN had strong binding affinities for the targets CHRM2,SLC6A4,PTGS2,and SLC6A2.Behavioral assessments demonstrated that PN significantly increased the sucrose preference index(P＜0.01,P＜0.001),reduced immobility time in the tail suspension and forced swimming tests(P＜0.01,P＜0.001),and enhanced exploratory behavior in the open field test(P＜0.05.P＜0.01,P＜0.001).PN significantly reduced the serum levels of inflammation markers(P＜0.05.P＜0.01,P＜0.001),as shown by enzyme-linked immunosorbent assay,and neurotransmitter analysis revealed that PN significantly increased the levels of serotonin and acetylcholine in the mouse hippocampus(P＜0.05).RT-qPCR showed that PN demonstrated the mRNA expression of SLC6A4(P＜0.05.P＜0.01,P＜0.001).Conclusions PN may improve depressive-like behavior in mice by modulating serotonin and acetylcholine levels,inhibiting inflammatory responses,participating in immune regulation,and exerting neuroprotective effects.

Objective Network pharmacology and molecular docking techniques were used to predict the potential mechanisms by which the active components of Piper nigrum(PN)regulate depressive-like behaviors in chronic restraint stress(CRS)mice.Methods The major chemical components and targets of PN were screened using the Traditional Chinese Medicine Systems Pharmacology database.Targets related to ferroptosis and depression were obtained from the Online Mendelian Inheritance in Man,GeneCards,and FerrDB databases.The intersecting targets were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analyses,and molecular docking was performed to validate the binding capacities between the core targets and their corresponding active components.Finally,we established a CRS mouse model.Mice were treated with PN 75,150,and 300 mg/kg for 4 weeks,followed by behavioral assessments and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)to verify the expression of core genes.Results Nine active components were screened from PN,corresponding to 27 targets,and 8377 targets related to depression and 547 targets associated with ferroptosis were screened from the databases.The intersection of these three sets resulted in 25 target genes.KEGG enrichment analysis revealed that these core targets were predominantly enriched in signaling pathways,including cholinergic synapses,serotonergic synapses,and neuroactive ligand-receptor interactions.Molecular docking result showed that the main active components of PN had strong binding affinities for the targets CHRM2,SLC6A4,PTGS2,and SLC6A2.Behavioral assessments demonstrated that PN significantly increased the sucrose preference index(P＜0.01,P＜0.001),reduced immobility time in the tail suspension and forced swimming tests(P＜0.01,P＜0.001),and enhanced exploratory behavior in the open field test(P＜0.05.P＜0.01,P＜0.001).PN significantly reduced the serum levels of inflammation markers(P＜0.05.P＜0.01,P＜0.001),as shown by enzyme-linked immunosorbent assay,and neurotransmitter analysis revealed that PN significantly increased the levels of serotonin and acetylcholine in the mouse hippocampus(P＜0.05).RT-qPCR showed that PN demonstrated the mRNA expression of SLC6A4(P＜0.05.P＜0.01,P＜0.001).Conclusions PN may improve depressive-like behavior in mice by modulating serotonin and acetylcholine levels,inhibiting inflammatory responses,participating in immune regulation,and exerting neuroprotective effects.

OBJECTIVE To optimize the heart-nourishing granule extraction process by network pharmacology and Box-Behnken response surface method. METHODS The active ingredients of heart-nourishing granule were excavated through network pharmacology and their mechanism of action in the treatment of coronary heart disease was preliminarily explored. Molecular docking technology was used to predict the binding ability of the active ingredients to the main targets. At the same time, based on the compatibility relationship between the monarch, minister, assistant and guide of the prescription, the pharmacological effects of the ingredients, and the content determination index components of each medicinal flavor in the 2020 edition of the Chinese Pharmacopoeia, the process evaluation index components of heart-nourishing granule were further determined. In addition, combined with the analytic hierarchy process to determine the weight of each component, the Box-Behnken response surface method was used to optimize the extraction process of heart-nourishing granule. RESULTS The main active components of heart-nourishing granule screened were catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin. By acting on core targets such as AKT1, IL-6, IL-1b, VEGFA, JUN and MAPK3, they regulated lipid and atherosclerosis, MAPK signaling and other pathways played a role in the treatment of coronary heart disease. Molecular docking results showed that the binding energies of active components and core targets predicted by network pharmacology were all < -5.0 kJ·mol-1. Five components catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin were calculated by analytic hierarchy process method. The weight coefficients of arginyl glycosides were 0.329 7, 0.329 7, 0.164 8, 0.109 9, and 0.065 9, respectively. The Box-Behnken response surface method obtained the optimal water extraction process: 10 times the amount of water was used to extract twice, and each time was decocted for 1.5 h. The verification test showed that the contents of the five components were consistent with the predicted values, and the RSD values were all <5%. CONCLUSION The extraction process of heart-nourishing granule based on network pharmacology and Box-Behnken response surface methodology is stable and feasible, which provided an experimental basis for its further quality improvement.

OBJECTIVE To establish a quality standard for rice-fired Glehnia littoralis . METHODS Appearance observation ， powder microscopic identification and thin-layer chromatography （TLC）identification were performed for the samples of rice-fired G. littoralis decoction piece. According to the relevant methods stated in 2020 edition of Chinese Pharmacopoeia （part Ⅳ），the contents of moisture ，total ash ，acid-insoluble ash ，water-soluble extract and acid-soluble extract were determined. The contents of psoralen，zanthoxylin，bergapten，imperatorin and isoimperatorin were determined by high performance liquid chromatography （HPLC）. RESULTS The rice-fired G. littoralis decoction pieces were round-like small segments ，slightly rough ，yellow（peeled） or dark yellowish brown （with peel ），special gas and slightly sweet taste. The powder was yellowish white. Under the microscope ， secretions and secretory cells ，ducts，gelatinized starch granules ，ray cells ，parenchyma cells ，etc. could be seen. TLC showed the spots developed clearly. In the chromatogram of the test sample ，there was the same blue fluorescent spot at the corresponding position of the chromatogram of isoimperatorin control sample. The moisture ，total ash ，acid-insoluble ash ，water-soluble extract and ethanol-soluble extract from 9 batches of samples were 5.82％-6.27％，3.19％-3.59％，0.21％-0.27％，24.91％-30.30％ and 20.66％ -25.83％ ，respectively. The linear range of psoralen ，zanthotoxin，bergapten，imperatorin and isoimperatorin were 0.240-2.400，0.320-3.200，0.224-2.240，0.292-2.920，0.208-2.080 µg/mL（all r＞0.999 0）. Limits of quantitation were 0.032 0， 0.030 0，0325 0，0.032 0，0.045 0 µg，respectively. Limits of detection were 0.100 8，0.089 6，0.071 5，0.090 0，0.132 0 µg， respectively. RSDs of prescision ，stability（24 h）and reprodu- cibility tests were less than 3％. Average recoveries were 100.56％ （RSD＝1.36％ ，n＝6），100.73％（RSD＝2.25％ ，n＝6）， 100.36％（RSD＝0.98％，n＝6），98.24％（RSD＝0.40％，n＝6） E-mail：853063968@qq.com and 99.40％（RSD＝0.35％，n＝6），respectively. The contents of above five components were 5.85-13.31，8.63-33.38，6.23- E-mail：shixiaofeng2005@sina.com 15.25，6.12-12.98，5.52-10.77 µg/g，respectively. The total contents were 34.20-83.47 µg/g. CONCLUSIONS It is preliminarily proposed that the moisture ，total ash and acid-insoluble ash should not exceed 7.30％，4.10％，0.30％. The water-soluble extract and ethanol-soluble extract are no less than 21.00％ and 18.00％，respectively. The total content of coumarin should not be less than 52.03 µg/g（with peel ）and 26.34 μg/g（peeled）. Established quality standard can be used for the quality control of rice-fired G. littoralis .

OBJECTIVES: To determine the potential molecular mechanisms underlying the therapeutic effect of curcumin on hepatocellular carcinoma (HCC) by network pharmacology and experimental in vitro validation. METHODS: The predictive targets of curcumin or HCC were collected from several databases. the identified overlapping targets were crossed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) platform. Two of the candidate pathways were selected to conduct an experimental verification. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium (MTT) assay was used to determine the effect of curcumin on the viability of HepG2 and LO2 cells. The apoptosis and autophagy of HepG2 cells were respectively detected by flow cytometry and transmission electron microscopy. Besides, western blot and real-time polymerase chain reaction (PCR) were employed to verify the p53 apoptotic pathway and adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK) autophagy pathway. HepG2 cells were pretreated with pifithrin-‍α (PFT-‍α) and GSK690693 for further investigation. RESULTS: The 167 pathways analyzed by KEGG included apoptosis, autophagy, p53, and AMPK pathways. The GO enrichment analysis demonstrated that curcumin was involved in cellular response to drug, regulation of apoptotic pathway, and so on. The in vitro experiments also confirmed that curcumin can inhibit the growth of HepG2 cells by promoting the apoptosis of p53 pathway and autophagy through the AMPK pathway. Furthermore, the protein and messenger RNA (mRNA) of the two pathways were downregulated in the inhibitor-pretreated group compared with the experimental group. The damage-regulated autophagy modulator (DRAM) in the PFT-‍α-pretreated group was downregulated, and p62 in the GSK690693-pretreated group was upregulated. CONCLUSIONS Curcumin can treat HCC through the p53 apoptotic pathway and the AMPK/Unc-51-like kinase 1 (ULK1) autophagy pathway, in which the mutual transformation of autophagy and apoptosis may occur through DRAM and p62.
AMP-Activated Protein Kinases/pharmacology* ; Apoptosis ; Carcinoma, Hepatocellular/pathology* ; Curcumin/pharmacology* ; Humans ; Liver Neoplasms/pathology* ; Network Pharmacology ; Tumor Suppressor Protein p53/metabolism*

Objective:To investigate the therapeutic effect of dorsal skin-tightening technique on the correction of mild penile curvature in children with hypospadias.Methods:The clinical characteristics of hypospadias patients (95 cases) with mild penile curvature (<30°) after degloving penis during operation in our hospital from Jan 2017 to Sep 2020 were analyzed retrospectively. Group A: A new technique, reconstructing penile pubic angle at 12 o'clock position of penile dorsal side after degloving and suturing forskin outer and inner plate with tension at 12 o'clock position, was performed in dorsal skin-tightening group (43 cases). Gtoup B: while in dorsal midline tunica albuginea plication group (52 cases), Buck fascia was incised on dorsal midline area, following by tunica albuginea plication with one or two stitches. The patients in Group A were 0.4 to 1.5 years old, and the median age was 1.1 years, urethral orifice were located on distal shaft in 36 cases, proximal in 7 cases.The patients in Group B were 0.5 to 2.6 years old, and the median age was 1.5 years, urethral orifice were located on distal shaft in 41 cases, proximal in 11 cases. The penile ventral curvature degree was recorded during regular follow-up (postoperative 6 and 12 months), as well as postoperative complications.Results:Artificial erection test showed penile curvature was corrected during surgery by measuring with protractor. There was no chordee by measuring with the side photos in all patients during an average of 1.6 years follow-ups. There were 4 case of urethral fistula in Group B and 3 cases in Group A. No cases of urethrostenosis, diverticulum or concealed penis was recorded. The difference of postoperative complications had no statistical significance.Conclusion:Hypospadias with mild penile curvature could be effectively corrected by dorsal skin-tightening technique, which showed a good result of early follow-up.

Objective:To explore the effect of different doses of dexmedetomidine (Dex) on levels of tight-junction protein claudin-1 and diamine oxidase (DAO) in patients undergoing laparoscopic radical resection of gynecological malignant tumors.Methods:A total of 60 patients with gynecological malignant tumors who were scheduled to undergo laparoscopic radical resection under general anesthesia from January 2019 to January 2020 in the Second Hospital of Shanxi Medical University were selected, including 43 cases of cervical cancer (stageⅠ-Ⅱ A), 9 cases of ovarian cancer (stageⅠ A-Ⅲ C), and 8 cases of endometrial carcinoma (stageⅠ). Accroding to the random number table method, the patients were divided into control group (group C), low-dose Dex group (group D 1) and high-dose Dex group (group D 2), with 20 cases in each group. Patients in group D 1 were given Dex 0.5 μg·kg -1·h -1 by constant rate intravenous infusion pump after induction until 30 min before the end of operation. Patients in group D 2 were given Dex 1.0 μg·kg -1·h -1 by constant rate intravenous infusion pump after induction until 30 min before the end of operation. Group C adopted the same calculation method and received the same amount of 0.9% sodium chloride solution by infusion pump. At 10 min before induction (T 1), 1 hour after pneumoperitoneum (T 2) and 12 hours after pneumoperitoneum (T 3), 5 ml of brachial vein blood was collected from the patients, and the levels of claudin-1 protein, DAO and blood glucose were measured. Results:At T 1, T 2 and T 3, the expression levels of claudin-1 in group C were (77.05±17.61) pg/ml, (66.76±12.97) pg/ml and (55.93±12.71) pg/ml, and the difference was statistically significant ( F = 10.449, P<0.05); the expression levels of DAO in group C were (4.83±0.93) ng/ml, (5.62±1.01) ng/ml and (5.98±1.21) ng/ml, and the difference was statistically significant ( F = 6.139, P < 0.05); the levels of blood glucose in group C were (4.82±0.66) mmol/L, (7.55±0.94) mmol/L and (6.51±0.54) mmol/L, and the difference was statistically significant ( F = 70.197, P < 0.05). At T 2, the expression level of claudin-1 in group D 1 was (69.12±13.02) pg/ml, which was not significantly different from group C ( t = -0.575, P > 0.05); the expression level of claudin-1 in group D 2 was (76.36±14.89) pg/ml, which was higher than that in group C, and the difference was statistically significant ( t = -2.175, P < 0.05). At T 3, the expression levels of claudin-1 in group D 1 and group D 2 were (66.14±14.36) pg/ml and (73.37±16.93) pg/ml, which were higher than that in group C, and the differences were statistically significant ( t values were -2.380 and -3.682, both P < 0.05). The expression levels of DAO in group D 1 and group D 2 were (5.02±0.84) ng/ml and (4.91±0.93) ng/ml at T 2, and (5.29±0.86) ng/ml and (5.20±0.98) ng/ml at T 3, which were lower than those in group C, and the differences were statistically significant ( t values were 2.051, 2.295, 2.079 and 2.285, all P < 0.05). The levels of blood glucose in group D 1 and group D 2 were (7.10±0.66) mmol/L and (6.77±0.97) mmol/L at T 2, and (5.95±0.94) mmol/L and (5.93±0.74) mmol/L at T 3, which were lower than those in group C, and the differences were statistically significant ( t values were 2.565, 5.374, 2.293 and 2.765, all P < 0.05). Conclusion:Continuous infusion of Dex can inhibit the stress response caused by long-term CO 2 pneumoperitoneum in laparoscopic radical resection of gynecological malignant tumors, and adjust the changes of expression levels of claudin-1 protein and DAO, reduce the damage of intestinal mucosal cells, facilitate the recovery of intestinal function, and the effect of high-dose Dex is better than low-dose Dex.

OBJECTIVE： To optimize the extraction technology of osthole from the pine needles of Cedrus deodara. METHODS： HPLC method was adopted to determine the content of osthole. Based on single factor test， ethanol volume fraction， extraction time and material-solvent ratio were selected as influential factors， and the content of osthole was selected as response value. Box-Behnken design-response surface methodology was used to optimize the extraction technology of osthole in pine needles of C. deodara. Validation test was conducted. RESULTS： The optimal extraction technology was as follows as ethanol volume fraction of 88％， material-solvent ratio of 1 ∶ 20 （g/mL）， extracting for 2 times， lasting for 57 min each time. Under this technology， average content of osthole was 0.675 7 mg/g （RSD＝1.78％， n＝3）， and the relative error of which to predicted value 0.680 9 mg/g was 0.59％. CONCLUSIONS： The optimal extraction technology is simple and feasible，and it can be used for the extraction of osthole from the pine needles of C. deodara.  

Objective: To explore the influence of plateau environment in the development and morphology of the main organs of the offsprings of migrated rats, and to observe the pathological changes of heart, brain, and lung tissues of the offsprings of migrated rats. Methods: The 8-week Wistar rats who lived in the plain area were migrated to the plateau area. After 1 week, a total of 56 female and male rats were fertilized with a ratio of 3:1, and all the pregnant rats were natural childbirth. The offsprings rat pups were divided into three groups: 1 month, 3 months, and 6 months. Ten offsprings (5 female and 5 male) were randomly selected in each group to collect the heart, brain, lung, liver, kidney and other major organs to measure the weights. In each group, 45 offsprings were randomly selected to conduct water maze test, open field test and test of captive reaction. The brain, heart, and lung tissues from 5 offsprings in each group were collected for tissue section and the pathological changes of above organ tissues were detected with HE staining. Results: The pregnant rats moved from the plain to the plateau had normal feeding behavior, without preterm birth or death. On average, the pregnant rats had 8 to 10 babies per litter, with a total of 345 offsprings. The weight gain of offspring was about 1. 0-1. 5 g per day. Some of the offsprings had low intake and difficulty in foraging, and 15 offsprings died 3-5 d after birth, with a mortality rate of 4. 3%. The weight and the ratio of liver weight to body weight of the 6-month-old offsprings were increased compared with the 1-month-old offsprings (F<0. 05). In Morris water maze test there were no drowning and death in 45 offsprings at different time points; the time to find the water platform of 7 offsprings in 3-month-old group was lower than the others (P<0. 05), but there were no significant differences between various groups at other time points (P>0. 05). In open field test, there were 6 offsprings in open fieled test in 1-month old group showed a longer stay time in the central region than other rats (P<0. 05); there were no significant differences between various groups at the other time points (P>0. 05). In spite of all the time points in the captive reaction experiment, all the animals behaved in a similar way. The HE staining results of myocardium tissue showed the myocardial inflammatory cell infiltration, venous congestion, nucleus disappearance, and visible cellular outline in the 1-3 month old offsprings and the myocardial vascular congestion and myocardial space enlargement in the 6-month-old offsprings. The HE staining results of brain tissue showed the glass body formation and nucleus disappearance in the 1-month-old offsprings and neuronal cell body deformation, blood vessel congestion and vacuolar degeneration in the 3-6-month-old offsprings. The HE staining results of lung tissue showed the thicker alveolar walls, lymphocyte and plasma cell infiltration, pulmonary capillary expansion and congestion in the 1-3-month-old offsprings and the thicker alveolar walls, lymphocyte and plasma cell infiltration, pulmonary capillary expansion and congestion, pulmonary interstitial edema and red blood cell liquefaction in the blood vessels in the 6-month-old offsprings. Conclusion: Tibetan plateau environment has an influence in the development and morphology of heart, brain and lung of the migrated rats, and the reason may be related to low pressure and hypoxia of plateau.

Objective To investigate the antibiotic resistance in clinical isolates in the Second Hospital of Hebei Medical University from 2015 to 2016.Methods A total of 16 292 strains of non-duplicate bacterial strains were collected.The isolates were subjected to identification and antimicrobial susceptibility testing on VITEK 2-Compact system.The data were processed and analyzed using WHONET 5.6 software.Results Specifically,7 961 and 8 331 strains of pathogens were collected in 2015,2016,respectively.Gram-negative bacteria accounted for 62.0％ in 2015 and 66.9％ in 2016,respectively.The top five pathogens isolated in these two years were still Klebsiella pneumoniae,Escherichia coli,Acinetobacter baumannii,Pseudomonas aeruginosa,and Staphylococcus aureus.The proportion of K.pneumoniae increased to the first place in 2016 which accounted for 16.1％.Coagulase negative Staphylococcus in blood samples decreased from 42.6％ in 2015 to 30.0％ in 2016.Vancomycin resistant strains were not found in Staphylococcus.In 2015 and 2016,the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) was 56.2％ and 51.3％,respectively.The prevalence of methicillinresistant coagulase negative Staphylococcus (MRCNS)was 79.3％ and 63.1％,respectively.Enterococcus faecalis and Enterococcus faecium accounted for 25.2％ and 73.2％ respectively in the 911 strains of Enterococcus.In 2015 and 2016,3.1％ and 2.9％ of the E.faecium strains were resistant to vancomycin,respectively.In 2016,the prevalence of carbapenemresistant strains increased in K.pneumoniae,E.coli and E.cloacae.K.pneumoniae showed increasing resistance rate to all the antimicrobial agents tested except for gentamicin and amikacin.The percentage of the K.pneumoniae strains resistant to imipenem and meropenem increased from 19.3％,18.5％ in 2015 to 24.2％,23.1％,respectively.A.baumannii isolates were still highly resistant to the commonly used antibiotics in these two years,but relatively susceptible to polymyxin B,tigecycline,cefoperazone-sulbactam and minocycline (＜30％ resistant).P.aeruginosa isolates showed lower resistance rate to amikacin (11.7％),cefoperazonesulbactam (15.5％),piperacillin-tazobactam (18.7％),ceftazidime (20.1％),cefepime (21.9％).P.aeruginosa presented a trend of declining resistance to all the antimicrobial agents tested from 2015 to 2016,except aztreonam,to which the resistant P.aeruginosa strains increased from 27.0％ in 2015 to 34.7％ in 2016.Conclusions The antimicrobial susceptibility profile of clinical bacterial isolates has been changing constantly.We need to adopt effective infection prevention and control measures in hospital and further standardize and control the use of antibacterial agents.