ObjectiveTo explore the effect of different drying conditions on the appearance and intrinsic quality indicators of Pinelliae Rhizoma for screening suitable drying conditions， so as to provide reference for its standardized production and quality evaluation. MethodsDifferent dried samples of Pinelliae Rhizoma were prepared by lime-assisted sweating method and intermittent drying method. Visual analysis was employed to measure the color brightness values（L^*） of the surface， cross-section and powder of the samples， texture analyzer was used to determine the hardness of the samples under different drying conditions. The total starch content was calculated by measuring the contents of amylose and amylopectin in the samples with ultraviolet-visible spectroscopy. High performance liquid chromatography（HPLC） was used to determine the contents of seven nucleoside components（uracil， hypoxanthine， uridine， inosine， guanosine， β-thymidine and adenosine） in the samples. Pearson correlation analysis was conducted to explore the correlation between the external characteristics and intrinsic indicators of the different dried samples. Principal component analysis（PCA） was used to comprehensively rank the data of various indicators， and partial least squares-discriminant analysis（PLS-DA） was used to screen differential components with variable importance in the projection（VIP） value>1. Furthermore， the difference between the optimal drying condition for Pinelliae Rhizoma and the traditional sun-drying method was explored by independent samples t-test. ResultsWith the increase of temperature， the color of the intermittently dried samples gradually deepened， while their hardness gradually decreased. Concurrently， the contents of extract， total starch， uridine and adenosine exhibited an upward trend， whereas the contents of uracil， hypoxanthine and inosine displayed a downward trajectory. Compared with the intermittent drying group， the content of extract in the samples subjected to lime-assisted sweating increased. With the increase of lime dose， the hardness and the total content of nucleoside components in the samples showed a downward trend， while the total starch content showed an upward trend. Correlation analysis showed that the comprehensive score of L^* was negatively correlated with the contents of uracil， hypoxanthine and inosine， and positively correlated with the contents of uridine， guanosine and adenosine. Hardness was negatively correlated with adenosine content， and positively correlated with the contents of inosine， uracil and hypoxanthine. Through comprehensive consideration and comprehensive score of principal components， the method of 5% lime-mixed sweating for 6 days emerged as the top-ranking approach. Except for the extract， the results of independent samples t-test showed that there was no significant difference between the 5% lime-mixed sweating for 6 days and the traditional sun-drying in terms of other content indicators. ConclusionThe whiteness and firmness of Pinelliae Rhizoma exhibit significant correlations with its chemical composition， while uridine， uracil， guanosine， adenosine and inosine are the key constituents responsible for the quality difference of Pinelliae Rhizoma under different drying conditions. The lime-assisted sweating method optimized in this study can be proposed as a viable alternative to the traditional sun-drying method. This method not only ensures the quality of the medicinal material but also effectively reduces the drying time and prevents mold contamination， which provides a valuable reference for the standardization of drying conditions and the establishment of quality evaluation criteria for Pinelliae Rhizoma.

In this paper， by consulting the historical herbs and medical classics coupled with related literature in modern research， the historical edition of Chinese Pharmacopoeia and local chronicles of traditional Chinese medicine （TCM） along with the ancient historical evolution of the processing methods of Pinelliae Rhizoma origin as well as the related processing methods of Pinelliae Rhizoma origin from 1959 to 2020 were systematically collated and summarized. It was found that the main processing methods of Pinelliae Rhizoma origin were peeling， decoction washing， lime wrapping and sun-drying. However， stacking， peeling， sun-drying or oven-drying are the primary methods in modern local chronicles of TCM. Meanwhile， washing， peeling， removing fibrous roots and sun-drying are the main methods in Chinese Pharmacopoeia. In addition， there were some changes in the quality evaluation of Pinelliae Rhizoma in different historical periods. Round and white were the best in the quality evaluation of Pinelliae Rhizoma in ancient times， while the evaluation indexes were further refined to size， color， texture， powder property， purity and evenness in modern herbal works. In modern studies， the quality of Pinelliae Rhizoma was mostly evaluated by the chemical components such as alkaloids， total organic acids， polysaccharides， nucleosides， fingerprint and pharmacodynamics. At present， the purification and drying stages of Pinelliae Rhizoma are in the transitional stage between the traditional manual peeling and natural drying methods as well as the modern mechanized and large-scale production. Therefore， a reasonable and feasible modern processing methods and guiding standards of Pinelliae Rhizoma are developed urgently to normalize the processing of Pinelliae Rhizoma and ensure the quality of medicinal materials.

Alfalfa(Medicago sativa L.)is the most important legume forage crop worldwide with high nutritional value and yield.For a long time,the breeding of alfalfa was hampered by lacking reliable information on the autotetraploid genome and molecular markers linked to important agro-nomic traits.We herein reported the de novo assembly of the allele-aware chromosome-level genome of Zhongmu-4,a cultivar widely cultivated in China,and a comprehensive database of genomic variations based on resequencing of 220 germplasms.Approximate 2.74 Gb contigs(N50 of 2.06 Mb),accounting for 88.39％of the estimated genome,were assembled,and 2.56 Gb contigs were anchored to 32 pseudo-chromosomes.A total of 34,922 allelic genes were identified from the allele-aware genome.We observed the expansion of gene families,especially those related to the nitrogen metabolism,and the increase of repetitive elements including transposable elements,which probably resulted in the increase of Zhongmu-4 genome compared with Medicago truncatula.Population structure analysis revealed that the accessions from Asia and South America had rela-tively lower genetic diversity than those from Europe,suggesting that geography may influence alfalfa genetic divergence during local adaption.Genome-wide association studies identified 101 sin-gle nucleotide polymorphisms(SNPs)associated with 27 agronomic traits.Two candidate genes were predicted to be correlated with fall dormancy and salt response.We believe that the allele-aware chromosome-level genome sequence of Zhongmu-4 combined with the resequencing data of the diverse alfalfa germplasms will facilitate genetic research and genomics-assisted breeding in variety improvement of alfalfa.

Objective:To investigate the specific effect of minimally invasive hard channel intracranial hematoma crush suction flow on patients with cerebral hemorrhage, and to analyze the prognostic factors.Methods:A total of 80 patients with primary intracerebral hemorrhage who were treated in Bengbu First Peopl′s hospital from Aug. 2014 to Aug. 2018 were selected, with 52 males and 28 females, aged from 48 to 81 years and averaged (66.07±13.34) years. The prognosis of patients was assessed using the Bathel Index (BI), and the patients were divided into a control group (BI≥ 60 point considered a good prognosis, n=54) and an experimental group (BI<60 point considered a poor prognosis, n=26). Suspicious prognostic factors were formulated, including gender, age, average drainage time, average puncture needle indwelling time, average hospital stay, Glasgow score, time from injury to operation, bleeding volume, cause of disease, bleeding site, postoperative rebleeding to carry out univariate analysis between groups, and incorporate the difference items in univariate analysis into multivariate logistics regression analysis to explore related prognostic factors were compared by single factor analysis, and the difference items in the single factor analysis were included into the multivariate logistics regression analysis for the related prognostic factors exploring. Results:The hemorrhages was successfully cleared in all patients, and the average drainage time was (5.07±1.25) days. Mean needle retention time was (4.84 ± 1.37) d. Mean length of stay time was (15.26±1.44) days. Allpatients completed follow-up after operation. During 1 year of follow-up, 12 patients showed obvious brain atrophy, subdural broadening and effusion. All patients were followed up for 3 years, and their symptoms completely disappeared and their daily living ability recovered. No death was occured during the follow-up. According to the BI assessment results, the scores of 54 patients were ≥60, with an average score of (74.57±4.36). The scores of 26 patients were <60, with an average score of (48.24±5.12). The age, time from injury to operation and amount of blood loss of control group were significantly lower than those of experiment group, and Glasgow scores were significantly higher than those of experiment group, with statistically significant differences ( P<0.05). The percentage of patients in control group with infratentorial hemorrhage (14.81%) was significantly lower than that in experiment group (34.62%), and the rate of postoperative re-bleeding (1.85%) was significantly lower than that in experiment group (23.08%), with statistically significant differences ( P<0.05). Glasgow score was positively correlated with the prognosis of patients ( OR=3.449, P=0.005). The time from injury to operation was negatively correlated with the prognosis ( OR=0.580, P=0.023). Supratente hemorrhage was a protective factor for patient prognosis ( OR=3.813, P=0.024). Postoperative re-bleeding was correlated with the patients ( OR=19.963, P=0.008). A ROC curve was drawn with Glasgow score and the time from injury to operation. Glasgow score was selected as cut-off point at 7.5 point, with sensitivity of 96.30% and specificity of 88.46%. The time from injury to operation was selected as the cut-off point at 8.35 h, with sensitivity of 84.62% and specificity of 92.59%. Conclusion:Minimally invasive hard channel crush suction flow is effective and safe in the treatment of intracranial hematoma. The severity of cerebral hemorrhage, infratentorial hemorrhage and postoperative re-bleeding are prognostic factors.

OBJECTIVE：To study the effects of stilbene glucoside （TSG）on the proliferation and estrogen receptor （ER）of human breast cancer T- 47D cells ，and to explore its estrogen-like effect and potential mechanism. METHODS ：Taking ER positive human breast cancer T- 47D cells as subjects ，using β-estradiol（β-E2，1×10－8 mol/L）as positive control ，CCK-8 assay was used to detect the cell proliferation after treated with different concentrations of TSG （1×10－8，1×10－7，1×10－6，1×10－5，1×10－4 mol/L）for 24，48，72 h；the cell proliferation rate was calculated. Western blotting assay and RT-PCR methods were adopted to detect the protein and mRNA expression of ER-α and ER-β in cells after treated with low，medium and high concentrations of TSG （1×10－8， 1×10－6，1×10－4 mol/L）for 48 h. RESULTS ：After treated with different concentrations of TSG for 24，48，72 h，the cell proliferation rate of each administration group at each time point （except for β-E2 group at 48 h）increased significantly ，compared with blank group ；those of TSG groups （1×10－5，1×10－6，1×10－7 mol/L）were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. After treated with low ，medium and high concentrations of TSG for 48 h，protein and mRNA expression of ER-α and ER-β in cells were increased significantly，compared with blank group （P＜0.05 or P＜0.01）；protein expression of ER-β in TSG low concentration group ，mRNA expression of ER-α in TSG groups as well as mRNA expression of ER-β in TSG low and high concentration groups were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can induce the in vitro proliferation of T- 47D cells and exert estrogen-like effects by promoting protein and mRNA expression of ER-α and ER-β， which is stronger than that of β-E2 at a certain concentration.

Objective To analyze the epidemiological surveillance results of human brucellosis in Shanxi Province from 2012 to 2017,to know the epidemic status of brucellosis,and to provide evidence for prevention and control of brucellosis.Methods Incidence date and surveillance date of disease outbreaks in Shanxi Province from 2012 to 2017 were collected,the retrospective analysis method was used to analysis the "three distribution" of brucellosis,outbreak situation and the results of serological and pathogenic surveillance in the 4 surveillance stations.Results A total of 36 220 brucellosis cases were reported from 2012 to 2017,the average incidence was 16.62/100 000;8 540 brucellosis cases were reported in 2014,with incidence 23.53/100 000;a total of 23 197 cases of brucellosis were reported mainly in Datong,Shuozhou,Jinzhong and Xinzhou,accounting for 64.04％ of the province total.The onset was seasonal,and the peak of the epidemic was from March to August,accounting for 67.23％ (24 350/36 220).The brucellosis cases were mainly youth (23 084),male (28 317),farmers and herdsman (32 616).In the 4 surveillance stations of the brucellosis,39 140 cases were investigated,of which 10 536 cases did serological test,in which 585 were positive for Brucella (5.55％).The highest positive rate of serological test was 9.50％ (226/2 738) which was found in Tianzhen.A total of 626 samples carried out pathogen culture,in which 107 strains of brucellosis were detected,the detection rate was 17.09％,and 106 strains Brucella were melitensis biovar 3 of the total strains except 1 mutant.Conclusions The reported incidence in Shanxi Province is in a decline tendency,but the situation of brucellosis epidemic is still relatively serious.It is suggested that the surveillance work should be strengthened;the epidemic situation of brucellosis should be mastered in time and effectively controlled.

Objective To assess the efficacy and side effects of intravesical instillation of BCG after transurethral resection of the bladder tumor (TURBT) in non-muscle invasive bladder cancer (NMIBC) patients.Methods The clinical data of patients treated with BCG 120 mg per course induced perfusion or more after TURBT from December 2013 to October 2016 in 18 hospitals of northeast China region,were analyzed retrospectively.The first part,data of 106 patients with moderate,high-risk NMIBC were collected.A total of 83 patients were male,while the other 23 patients were female.The average age was 66.7 years old.The clinical staging were T1 in 86(81.1％) cases,Ta in 20(18.9％) cases and carcinoma in situ in 6 (5.7％) patients.Intravesical instillation of BCG was executed after transurethral resection of the bladder tumor.The incidence rate of recurrence and progression during more than 6 months' follow-up time were observed.Multivariate analyses were done by using logistic analysis and Cox proportional hazards regression model with Kaplan-Meier method.The second part,treatment compliance of 276 patients with bladder cancer,including moderate/high-risk NMIBC in 263 cases,moderate/high-risk NMIBC followed with renal pelvis/ureteral carcinoma in 8 cases were and moderate/high-risk NMIBC with renal pelvis/ureteral carcinoma in 5 cases who treated with BCG after the surgeries,were observed.Patients consisted of 211 males and 65 females with average age of 68.3 years.Results With a median follow-up of 12 months,9 (8.5％) patients experienced tumor recurrence and 2 (1.9％) patients were found progression in the first part.The one-year cancer free recurrence rate of the patients was 91.5％.Statistically significant prognostic factors for recurrence identified by multivariable analyses were prior recurrence of the tumors (OR =3.214,95％CI0.804-12.845,P =0.099).In the second port,an incidence rate of adverse effects was 64.1％ (177/276).The Ⅲ/Ⅳ degree complications were occurred in 11 patients and satisfactory outcomes achieved with active treatment.A total of 36 patients withdrawal with the major causes were recurrence and progression of bladder tumor in 12 cases (4.4 ％),9 cases (3.3 ％) with economic reasons and 11 cases (4.0％) with serious complications.Conclusions NMIBC patients treated with intravesical BCG therapy have approving cancer free recurrence rates and acceptable adverse effects.Prior recurrence may be prognostic factor of recurrence after intravesical BCG therapy.

OBJECTIVE： To investigate estrogen-like effect of tetrahydroxy stilbene glucoside （TSG） and its effects on the expression of estrogen receptor （ER） in uterus of sexually immature mice. METHODS： Totally 60 sexually immature Kunming mice were randomly divided into normal group， positive control group （estradiol valerate， 0.18 mg/kg）， TSG low-dose and high-dose groups （50， 150 mg/kg）， TSG low-dose and high-dose groups+estradiol valerate groups （same dose as medication alone group）. Normal group was given constant volume of water intragastrically， and administration groups were given relevant medicine 0.2    mL/10 g， once morning and night， for consecutive 5 d. The uterus index and body weight increase of mice in each group were determined and calculated the next day after the last administration. The contents of serum estrogen （E2， LH， FSH） were determined by ELISA. HE staining was used to observe the morphology characteristics of uterus， and uterine tube diameter and endometrial thickness were detected. The expression of ER（ER-α and ER-β） in uterus was detected by immunohistochemical staining. RESULTS： The myometrium of the mice in normal group was parallel and compact， the epithelium of the uterus was columnar， and the expression of ER-α and ER-β was low. The uterine tube diameter， endometrium and epithelium of mice in each administration group increased， thickened or proliferated in varying degrees， and the expression of ER-α and ER-β changed. Compared with normal group， uterus indexes （positive control group， TSG high-dose group， TSG+estradiol valerate groups）， the increase of body weight （positive control group， TSG high-dose groups， TSG low-dose+estradiol valerate group）， uterine tube diameter and endometrial thickness （positive control group， TSG low-dose group， TSG+estradiol valerate groups）， the expression of ER-α （positive control group， TSG+estradiol valerate groups） and the expression of ER-β （postive control group， TSG high-dose+estradiol valerate group）were increased significantly， while serum contents of LH （positive control group， TSG high-dose group） and FSH （TSG low-dose+estradiol valerate group） were decreased significantly （P＜0.05 or P＜0.01）. The uterus index， uterine tube diameter， endometrial thickness and the expression in ER-α and ER-β of TSG+estradiol valerate groups， the increase of body weight and serum content of E2 in TSG low-dose+estradiol valerate group were significantly higher than same TSG dose alone groups （P＜0.05 or P＜0.01）. The uterus index， uterine tube diameter， endometrial thickness and the expression of ER-α and ER-β in TSG groups， uterine tube diameter and the expression of ER-β in TSG+estradiol valerate groups， body weight increase of mice in TSG low-dose group were significantly lower than positive control group， while serum content of LH in TSG+estradiol valerate groups were significantly higher than positive control group （P＜0.05 or P＜0.01）. CONCLUSIONS： TSG can increase uterus indexes and body weight of sexually immature mice to certain extent， regulate estrogen level， increase the diameter of uterine tube and endometrial thickness and up-regulate the expression of ER in the uterus， showing certain estrogen-like effect， which is weaker than that of estradiol valerate. Combined use of them may antagonize the effect of estradiol valerate.

OBJECTIVE： To study the effects of processed Polygonum multiflorum containing serum on the proliferation and the expression of estrogen receptor （ER） of human breast cancer T-47D cells， and to investigate its phytoestrogen （PE）-like effect. METHODS： Sexually immature SD rats were randomly divided into estradiol valerate （Ev） group （positive control， 0.12 mg/kg）， processed P. multiflorum low-dose and high-dose groups （0.75， 3 g/kg， by crude drug）， low-dose and high-dose processed P. multiflorum+Ev groups （same dose as single drug group）， with 10 rats in each group. Blank group was given constant volume of water intragastrically， and administration groups were given relevant medicine intragastrically； once day and night， for consecutive 4 days. Two hours after last administration， blank serum and containing serum were prepared. T-47D cells were also randomly divided into blank group， Ev group， low-dose and high-dose processed P. multiflorum groups， low-dose and high-dose processed P. multiflorum+Ev groups， and then were cultured in medium which contained 20％ blank serum or drug containing serum. CCK-8 assay was used to detect proliferation rate （PR）. Western blotting assay and RT-PCR were used to detect the protein and mRNA expression of ER-α and ER-β. RESULTS： Compared with blank group， PR of administration groups [each administration group （24 h）， other administration groups （48， 72 h） except for high-dose processed P. multiflorum+Ev group] were increased significantly； high-dose processed P. multiflorum group （72 h） was significantly higher than Ev group， and low-dose processed P. multiflorum+Ev group （72 h） was significantly higher than the same-dose processed P. multiflorum group； high-dose processed P. multiflorum+Ev group （72 h） was significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. Relative protein expression of ER-α in Ev group， high-dose processed P. multiflorum group and low-dose processed P. multiflorum+Ev group， relative mRNA expression of ER-α and protein expression of ER-β in administration groups， relative mRNA expression of ER-β in Ev group， low-dose processed P. multiflorum group and processed P. multiflorum+Ev groups were all increased significantly. Relative protein and mRNA expression of ER-α in Ev group were significantly higher than processed P. multiflorum groups and combination groups. Relative protein and mRNA expression of ER-β in Ev group were significantly lower than low-dose processed P. multiflorum+Ev group， but relative mRNA expression of ER-β was significantly higher than processed P. multiflorum groups and high-dose processed P. multiflorum+Ev group. Relative protein and mRNA expression of ER-α and ER-β in low-dose processed P. multiflorum+Ev group as well as relative mRNA expression of ER-β in high-dose processed P. multiflorum+Ev group were significantly higher than the same-dose processed P. multiflorum group. Relative protein and mRNA expression of ER-α in high-dose processed P. multiflorum+Ev group were significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. CONCLUSIONS： The processed P. multiflorum containing serum can promote the proliferation of human breast cancer T-47D cells， and play the PE-like role through promoting protein and mRNA expression of ER-α and ER-β. However， the above effects are weaker than estrogen， and the combination of the two may antagonize the effect of estrogen.

OBJECTIVE: To investigate the differentially expressed microRNAs(miRNAs) in human embryonic lung fibroblast MRC-5 cells stimulated by transforming growth factor-β1(TGF-β1) using microarray chip, and screen for key genes and signaling pathways of fibroblast trans-differentiation. METHODS: The miRNA expression gene chip dataset GSE43992 on TGF-β1 stimulated MRC-5 cells were downloaded from high-throughput Gene Expression Omnibus(GEO) database of National Center for Biotechnology Information of the United States. The R language Limma package was used to screen the differentially expressed miRNAs. Corresponding target genes were predicted by miRWalk database performed by Gene Ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analysis. The protein-protein interaction(PPI) network was constructed by the search tool for the Retrieval of Interacting Genes database. RESULTS: A total of five differentially expressed miRNAs were identified, including four up-regulated miRNAs and one down-regulated miRNA; and 42 corresponding differentially expressed target genes were predicted. GO analysis indicated that the target genes were significantly enriched in collagen catabolic process, extracellular matrix organization, membrane organization, collagen fibril organization, and cellular response to amino acid stimulus. The results of KEGG pathway analysis showed that the signaling pathways corresponding to miRNAs and target genes were mainly concentrated in 18 signaling pathways, that were mainly related to the age-ethnic signaling pathways and protein digestion and absorption miRNAs in tumors and diabetic complications. The core genes transfected into the myofibroblasts by the three fibroblasts screened by the PPI network were threonine kinase 1, estrogen receptor 1 and β-catenin. CONCLUSION: Five differentially expressed miRNAs, 42 target genes, 18 signaling pathways, and 3 core genes related to TGF-β1-induced MRC-5 cell trans-differentiation were screened. It can provide new reference for the treatment and research of many diseases including pneumoconiosis and pulmonary fibrosis.