Objective To analyze the registered clinical trials of headache treated by TCM;To discuss the current research status;To provide reference for the optimization of subsequent clinical trial research plans.Methods All clinical trials of headache treated by TCM were retrieved from the ChiCTR and the ClinicalTrials.The retrieval time was from the database establishment to May 22,2023.The general characteristics,study types,intervention measures and outcome indicators of the trials were analyzed respectively.Results A total of 104 registered studies were included,with the number of registered studies increasing since 2004 and reaching a peak in 2020,involving 25 provincial administrative regions or countries and 69 clinical trial institutions;the funding sources were mainly scientific research funds of universities,national finance and local finance.The research type was mainly intervention research;the designing scheme was mainly randomized parallel control study;the high frequency random method was simple random method;45 registered studies used blind methods.Exploratory studies/pre-trials were the most commonly used in the phases of clinical researches.Most of the registered studies were single-center clinical trials with a total sample size of 9 648 patients.The main interventions were acupuncture and oral Chinese medicines.The high frequency outcome indicators included life quality of score,headache attack frequency,headache attack days and headache severity,etc.There were some problems in outcome indicators,such as non-standard,lack of TCM characteristic advantages,and insufficient patient participation.Conclusion The number of registered studies of headache treated by TCM has increased by year,but there are some problems in design elements,such as random method,blind method,number of research centers,sample size and the setting of outcome indicator.

Migraine is a common neurovascular disorder manifested by recurrent severe headaches on one or both sides， occasionally accompanied by nausea， vomiting， photophobia， and phonophobia. It has the characteristics of recurrent attacks and family inheritance. Traditional Chinese medicine （TCM） believes that migraine belongs to the category of "head wind"， which is mostly caused by external wind and is related to the internal stirring of liver wind. Sanpiantang comes from the Record of Syndorme Differentiation·Headache （Bianzhenglu·Toutongmen） created by the physician CHEN Shiduo of the Qing Dynasty. It is composed of Chuanxiong Rhizoma， Angelicae Dahuricae Radix， Pruni Semen， Cyperi Rhizoma， Bupleuri Radix， White Mustard Seed， and Glycyrrhizae Radix et Rhizoma， with the functions of moving Qi to release pain， activating blood and resolving stasis， which is commonly used for the treatment of migraine in clinic. Current clinical studies on the application of Sanpiantang to the treatment of migraine mostly used modified Sanpiantang， either alone or in combination with western medicine/acupuncture. The results of these clinical trials showed that Sanpiantang could significantly lower migraine score， pain visual analog scale and endothelin level， reduce the frequency of painkiller use， and remarkably alleviate migraine symptoms， with few side effects. The animal experiments focused on exploring the mechanism of action of modified Sanpiantang from different anatomical levels of migraine， which mainly included reducing nitric oxide （NO） and nitric oxide synthase （NOS）， reduceing the release of neurotransmitters such as 5 -hydroxyline (5-HT) and neurotipides (NPY)， suppressing neuronal excitation， and blocking the transmission of nociceptive pathways， thereby promoting cerebral blood flow， regulating neurotransmitters and preventing migraine. Based on the pathogenesis of migraine， this paper systematically reviewed the latest progress in clinical application and experimental research of modified Sanpiantang， and summarized its mechanism of action of preventing and treating migraine， which provided new ideas for clinical treatment of migraine.

ObjectiveTo observe the behavioral and pain threshold alterations， as well as the changes in indexes related to depression and pain in the serum and central system in mice stressed by maternal separation and chronic neuropathic pain， and explore the underlying mechanism of Wenyang prescription （WY）， Jieyu prescription （JY）， and Wenyang Jieyu prescription （WYJY） in improving depression and pain sensitivity. MethodThe birth date of mice was recorded as PD0. After birth， the mice were divided into a blank group and an experimental group. The neonatal mice in the experimental group underwent maternal separation in PD5-14 at 8 h·d^-1. After ablactation， the mice were divided into a maternal separation group， a WY group （Erxian decoction， 5.84 g·kg^-1）， a JY group （Xiaoyaosan， 12.00 g·kg^-1）， a WYJY group （16.68 g·kg^-1）， and a fluoxetine group （2.60 mg·kg^-1）， with 15 mice in each group. Meanwhile， 15 male mice of the same age without maternal separation were assigned to the normal control group. Mice in the blank group and the maternal separation group were fed on a regular chow diet in PD21-PD90， while the remaining groups were fed on the corresponding drugs. In PD91， sciatic nerve ligation was performed to induce a model of maternal separation and chronic neuropathic pain. The open field test was used to observe the depression-like behaviors of mice in each group， and the mechanical and temperature pain thresholds were measured to detect the pain sensitivity of mice in each group. The serum levels of corticosterone （CORT）， substance P， and β-endorphin （β-EP） were determined by enzyme-linked immunosorbent assay （ELISA）， and the expression of the glucocorticoid receptor （GR） in the amygdala and β-EP protein in the hypothalamus was detected by immunohistochemistry. The mRNA expression levels of amygdala GR gene （Nr3c1）， FK506 binding protein 5 gene （FKBP5）， metabolic glutamate receptor 5 gene （GRM5）， and brain-derived neurotrophic factor （BDNF） were detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultCompared with the blank group， the maternal separation group showed reduced stay time and total distance traveled in the 5-min open field test （P<0.01）， reduced mechanical pain threshold （P<0.01）， increased serum CORT and β-EP （P<0.01）， declining FKBP5 mRNA expression （P<0.01）， and increased hypothalamic β-EP expression （P<0.05）. Compared with the maternal separation group， the groups with drug intervention showed prolonged stay time （P<0.05， P<0.01） and up-regulated pain thresholds to different degrees. The total distance traveled in the 5-min open field test increased in the WY group， the WYJY group， and the fluoxetine group （P<0.05， P<0.01）. The JY group showed decreased serum CORT （P<0.01）， reduced β-EP ， and increased BDNF mRNA （P<0.01）. Nr3c1 and GRM5 mRNA decreased in the WY group （P<0.05， P<0.01）. The WYJY group showed decreased serum CORT （P<0.05）and decreased Nr3c1， GRM5， and BDNF mRNA （P<0.05， P<0.01）. The levels of β-EP expression were elevated to different degrees in the groups with drug intervention， but the differences were not significant. The levels of GR expression in the WY group， the JY group， and the WYJY group increased （P<0.05）. ConclusionWYJY can inhibit central pain sensitization and regulate hypothalamic-pituitary-adrenal gland （HPA） axis function by enhancing the expression of GR in the amygdala and inhibiting neuroplasticity and excitability in the amygdala to relieve depression-like behaviors and improve somatic hyperalgesia.

Objective:To evaluate the effects of Xiongzhi Dilong decoction(XZDLD)and its wind medicine on calcitonin gene-related peptide(CGRP)-induced migraine and explore the mechanism through the CGRP/inducible nitric oxide synthase(iNOS)pathway.Methods:Rats were divided into control,model,XZDLD,XZDLD(external wind),XZDLD(internal wind),and olcegepant groups.CGRP was injected into the dura mater to induce a migraine.The frequency of head scratching,cage climbing,and facial grooming was observed.The pain threshold,the levels of CGRP,pituitary adenylate cyclase activating polypeptide(PACAP),substance P(SP),and the plasma protein extravasation(PPE)ratio were measured.The phosphorylation levels of p38,extracellular signal-regulated kinase 1/2(ERK1/2),and expression of iNOS were detected by western blot.Results:Compared with the model group,the three modified XZDLD groups showed reduced frequency of head scratching and cage climbing in the first 30 min(all P＜.05).Facial grooming frequency was reduced in XZDLD and XZDLD(external wind)groups(P=.0003 and P=.0131,respectively).External wind medicine played a more important role in increasing mechanical pain threshold than internal wind medicine.Moreover,compared with the model group,the three modified XZDLD groups demonstrated reduced plasma levels of CGRP and PACAP(all P＜.05).No difference in the SP level was observed among the six groups.XZDLD reduced PPE ratio.XZDLD and XZDLD(external wind)groups suppressed the CGRP/iNOS pathway by inhibiting the p-p38/p38 ratio and the expression of iNOS.No difference in pERK1/2/ERK1/2 ratio was detected among the six groups.Conclusion:XZDLD increases pain threshold,downregulates the expression of CGRP and PACAP,and reduces PPE ratio by inhibiting the CGRP/iNOS pathway.External wind medicine is more effective than internal one on improving facial grooming and head scratching,increasing the mechanical pain threshold,and inhibiting the expression of iNOS.

OBJECTIVETo investigate the drug resistance, β-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding genes of, and to explore its structure and pathogenic function.

METHODSThe strain was isolated by plate streaking method and identified by automatic bacteria detection system and 16S RNA gene PCR. Microdilution method was applied for drug susceptibility test. β-lactamases, extended spectrum β-lactamases (ESBL) and carbapenemases were detected using nitrocefin-disk, Kirby-Bauer disk, and Hodge test, respectively. Five β-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene of the isolate were amplified by PCR for sequencing. Bioinformatic softwares were used to analyze the structure and function of the product of protoporphyrin ferrochelatase-encoding gene.

RESULTSA strain belonging towas isolated. This isolate was sensitive to cefepime, ciprofloxacin, ofloxacin and tigecycline, but resistant to five penicillins, four cephalosporins and two carbapenems antibiotics. The isolate produced β-lactamases but did not produce ESBL and carbapenemases. The isolate had five distinct β-lactamase-encoding genes and protoporphyrin ferrochelatase-encoding gene. The product of protoporphyrin ferrochelatase-encoding gene contained two functional domains of protoporphyrin ferrochelatase belonging to type Ⅱ chelatase superfamily that presented the most closely genetic relationship with the protoporphyrin ferrochelatase of.

CONCLUSIONSThe isolate ofhas a higher resistance to β-lactam antibiotics and its β-lactamase-encoding genes are different with the common bacterial β-lactamase-encoding genes. Protoporphyrin ferrochelatase may act as an important virulence factor of.

Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Objective To investigate the changes of non-linear dynamics characteristics of electroencephalogram(EEG) in patients with the stagnation of Liver-Qi syndrome.Methods 15 Liver-Qi stagnation syndrome patients and 15 normal persons were paired by sex and age.The data of EEG under three states(eyes closed,eyes opened,and mental arithmetic with eyes opened) were analyzed by the parameter of point-wise correlation dimension(PD2).Results PD2 of the stagnation of Liver-Qi syndrome group increased significantly compared with that of the healthy control group and always maintain high level.The difference of PD2 between two groups was gradually reduced along with the increase of stimulated condition.Conclusion Non-linear dynamic analysis is more appropriate for the study of mental functions changes of stagnation of Liver-Qi syndrome and can be used to investigate the brain mechanism of the stagnation of Liver-Qi syndrome.

Objective To explore the characteristics of patients with stagnation of liver-qi syndrome using brain functional imaging techniques.Methods10 patients with stagnation of liver-qi syndrome and 20 normal persons whom paired by sex and age were scanned with positron emission tomography(PET).Data analysis was carried out using SPM in MATLAB software.ResultsCompared with normal,some cerebral areas of the patients with stagnation of liver-qi syndrome showed glucose metabolism letdown(P<0.001),including right parietal lobe(postcentral gyrus,BA1/2),temporal lobe(middle temporal gyrus,BA37),frontal lobe(middle frontal gyrus,BA9/10)etc.,while some cerebral areas showed glucose metabolism heighten(P<0.001)including right frontal lobe(superior frontal gyrus,inferior fontal gyrus,BA22/28/44),right cingulated gyrus,double midbrain and cerebella,etc.ConclusionThe function of some cerebral areas can be changed undergoing stagnation of liver-qi syndrome.

Objective To study the personality characteristics of the stagnation of liver-qi syndrome cases with Minnesota Multiphasic Personality Inventory (MMPI).Methods 20 liver-qi stagnation syndrome cases and 20 normal persons whose paired by sex and age were tested by MMPI. The scores of 13 clinical scales and the section plane of MMPI were acquired and analyzed. Results There was a significant difference in 9 clinical scales (F, Pa, D, Pt, Si, Sc, Hs, Hy, Pd) between the liver-qi stagnation syndrome cases and normal persons ( P<0.05～0.001). The scores of Hs, D, Hy, Pd in the liver-qi stagnation syndrome group were higher than the Chinese normal model. The MMPI section plane of liver-qi stagnation group presented the type of 3/1. Conclusion The patients with liver-qi stagnation have some special personality characteristics such as depression, anxiety, loneliness, indifferent and attention.

Objective:To explore the effect of Naoshuning on the protein expression of Matrix metalloproteinases(MMPs) in experimental injuried brain tissue of rats. Methods:Immunohistochemistry was used to determine the changes of protein expression of MMPs. Brain tissue water content,permeability and ultramicrostructure of blood-brain barrier(BBB) were also observed. Results:Compared with the sham group,the brain tissue water and EB content of injured side and the level of MMP-2 and MMP-9 protein expression in brain tissue around contusion in model group increased obviously(all P