Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

OBJECTIVE: In Pakistan, traditional medicines are an important component of the medical system, with numerous varieties and great demands. However, due to the scattered resources and the lack of systematic collection and collation, adulteration of traditional Pakistani medicine (TPM) is common, which severely affects the safety of their medicinal use and the import and export trades. Therefore, it is urgent to systematically organize and unify the management of TPM and establish a set of standards and operable methods for the identification of TPM. METHODS: We collected and organized the information on 128 TPMs with regard to their medicinal parts, efficacy, usage, and genetic material, based on Pakistan Hamdard Pharmacopoeia of Eastern Medicine: Pharmaceutical Codex. The genetic information of TPM is summarized from national center for biotechnology information (NCBI) and global pharmacopoeia genome database (GPGD). Furthermore, we utilized bioinformatics technology to supplement the chloroplast genome (cp-genome) data of 12 TPMs. To build the web server, we used the Linux + Apache + MySQL + PHP (LAMP) system and constructed the webpage on a PHP: Hypertext Preprocessor (PHP) model view controller (MVC) framework. RESULTS: We constructed a new genomic database, the traditional Pakistani medicine genomic database (TPMGD). This database comprises five entries, namely homepage, medicinal species, species identification, basic local alignment search tool (BLAST), and download. Currently, TPMGD contains basic profiles of 128 TPMs and genetic information of 102 TPMs, including 140 cytochrome c oxidase subunit I (COI) sequences and 119 mitochondrial genome sequences from Bombyx mori, 1 396 internal transcribed spacer 2 (ITS2) sequences and 1 074 intergenic region (psbA-trnH) sequences specific to 92 and 83 plant species, respectively. Additionally, TPMGD includes 199 cp-genome sequences of 82 TPMs. CONCLUSION TPMGD is a multifunctional database that integrates species description, functional information inquiry, genetic information storage, molecular identification of TPM, etc. The database not only provides convenience for TPM information queries but also establishes the scientific basis for the medication safety, species identification, and resource protection of TPM.

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.

OBJECTIVE：To i nvestigate the spectrum-effect relationship of analgesic and anti-inflammatory effects of ethyl acetate extract from Zhuang medicine Stahlianthus involucratus from different habitats. METHODS ：Ten batches of S. involucratus from different habitats were used as samples to investigate the anti-inflammatory and analgesic activities of ethyl acetate extracts by xylene induced ear swelling test and acetic acid induced writhing test in mice. HPLC fingerprints of 10 batches of ethyl acetate extract from S. involucratus were established and their similarity was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition），and the common peaks were identified by comparison with the control. The spectrum-effect relationship of anti-inflammatory and analgesic effects of ethyl acetate extract from S. involucratus were analyzed on the basis of Pearson correlation coefficient （auricle swelling degree and writhing times in 15 min as pharmacodynamic indexes ）and Grey relational analysis （inhibition rate of ear swelling and analgesic rate as pharmacodynamic indexes ）. RESULTS ： batches of ethyl acetate extract from S. involucratus had obvious anti-inflammatory and analgesic effects ； inhibition rates of ear swelling in mice were 46.43％-55.16％，and the analgesic rates of mice were 45.56％-52.72％. A total of 18 common peaks were identified in 10 batches of samples ，andthe similarity between them and the control fingerprint was 0.994-0.997. Compared with substance control ，the pea ks 1，2 and 4 were identified as protocatechuic acid ， p-hydroxy- 0771-4953513。E-mail：liangjie1101@126.com benzoic acid and p-hydroxybenzaldehyde，respectively. Results of Pearson correlation analysis showed that peak 10 and peak 18 were significantly negative correlated with auricle swelling degree and writhing times in 15 min（r were values －0.853，－0.738，P values were 0.002，0.015，respectively）. Results of Gray correlation degree analysis showed that the correlation degree of 18 common peaks with inhibition rate of ear swelling and analgesic rate were all greater than 0.65；among them ，peaks 14，1（protocatechuic acid ），17，9，4（p-hydroxybenzaldehyde），2（p-hydroxybenzoic acid ）， 16，7 and 6 showed the relatively high correlation degree （correlation degree ＞0.7）；peak 1（protocatechuic acid ），17，14，9，16，2 （p-hydroxybenzoic acid ）and 4（p-hydroxybenzaldehyde）showed the relatively high correlation degree （correlation degree ＞0.7）. CONCLUSIONS：The ethyl acetate extract of S. involucratus show good anti-inflammatory and analgesic effects. Peak 1 （protocatechuic acid ），2（p-hydroxybenzoic acid ），10，14，17，18 may be its main active ingredients.

Objective:To investigate the improvement and clinical effect of lip depilation with 810 nm semiconductor laser.Methods:Retrospective analysis was performed on female labial hair removal patients admitted to Bethune Hospital in Shanxi from January 2017 to October 2018. The patients who visited the clinic from January 2018 to October 2018 were taken as the experimental group, and those who visited the clinic from January 2017 to December 2017 were taken as the control group. A combination model of super hair removal(SHR) + hair removal(HR) was adopted to perform the treatment with 810 nm YfTON semiconductor freezing point laser hair removal. The experimental group was treated with surface anesthetic and SHR mode dumbbell sliding mode 8 J/cm 2+ HR mode with high energy (starting from the upper limit of reference energy value), and the patients were treated with local drum and chin coordination. In the control group, no surface anesthetic was applied, and SHR mode was used for linear round-trip sliding + HR mode for reference energy treatment. After the third and fifth treatment, the patients in the two groups were treated with a hair size of 1 cm×1 cm on one side of the mouth corner with heavy hair. A hair analyzer was used to count the number of similar hairs visible to the naked eye at a social distance of about 1 meter. The final hair removal rate was calculated as excellent (70%-90%, including 70%), as improvement (40%-70%, including 40%), as invalid (<40%). The depilation rate after 3 and 5 treatments was compared between the 2 groups, the number of treatments was analyzed, and the recurrence rate and comfort level of the patients were followed up 6 months after treatment. Wilcoxon rank sum test was used to compare the measurement data. Comparison of counting data was tested, and P<0.05 was considered statistically significant. Results:A total of 200 patients were included in this study, including 100 in the experimental group, with an age of (28.58±6.01) years. The control group included 100 patients (28.57±6.07) years old. The skin type of patients was performed according to Fitzpatrick and Chinese characteristics Ⅱ-Ⅳ. After statistical test, there were no statistically significant differences in age and skin type between the two groups. After half a year of follow-up, the average number of treatment times in the experimental group was 5(5, 6) times, lower than that in the control group 7(6, 8) times( T=-9.279, P<0.001). The three and five depilation rates of the experimental group were[70.0%(68.0%, 74.8%), 86.0%(83.0%, 88.0%)], and the final depilation rates of the experimental group were 89.0%(86.7%, 90.0%), all higher than those in the control group[40.0%(37.0%, 42.0%), 72.0%(70.0%, 74.0%), 86.5%(85.0%, 88.0%)]. The differences between the two groups were statistically significant ( T3=43.986, T5=25.151, Tfinal=5.547, all P< 0.001). The intermittent times of single sliding treatment in the experimental group ≤2 times, compared with the control group > 2 times, the comfort level was improved. No blisters or coloration occurred in any of the patients. Patients in the 2 groups were followed up for half a year after the last hair removal. In the 2 groups, there were sparse new fine hairs and soft hair growth that were not included in the hair count, and the included hairs were thinner than those before the treatment. Conclusions:Compared with the control group, the treatment cycle is shorter, the efficiency is higher and the comfort level of the patients is higher in the experimental group.

Objective:To investigate the improvement and clinical effect of lip depilation with 810 nm semiconductor laser.Methods:Retrospective analysis was performed on female labial hair removal patients admitted to Bethune Hospital in Shanxi from January 2017 to October 2018. The patients who visited the clinic from January 2018 to October 2018 were taken as the experimental group, and those who visited the clinic from January 2017 to December 2017 were taken as the control group. A combination model of super hair removal(SHR) + hair removal(HR) was adopted to perform the treatment with 810 nm YfTON semiconductor freezing point laser hair removal. The experimental group was treated with surface anesthetic and SHR mode dumbbell sliding mode 8 J/cm 2+ HR mode with high energy (starting from the upper limit of reference energy value), and the patients were treated with local drum and chin coordination. In the control group, no surface anesthetic was applied, and SHR mode was used for linear round-trip sliding + HR mode for reference energy treatment. After the third and fifth treatment, the patients in the two groups were treated with a hair size of 1 cm×1 cm on one side of the mouth corner with heavy hair. A hair analyzer was used to count the number of similar hairs visible to the naked eye at a social distance of about 1 meter. The final hair removal rate was calculated as excellent (70%-90%, including 70%), as improvement (40%-70%, including 40%), as invalid (<40%). The depilation rate after 3 and 5 treatments was compared between the 2 groups, the number of treatments was analyzed, and the recurrence rate and comfort level of the patients were followed up 6 months after treatment. Wilcoxon rank sum test was used to compare the measurement data. Comparison of counting data was tested, and P<0.05 was considered statistically significant. Results:A total of 200 patients were included in this study, including 100 in the experimental group, with an age of (28.58±6.01) years. The control group included 100 patients (28.57±6.07) years old. The skin type of patients was performed according to Fitzpatrick and Chinese characteristics Ⅱ-Ⅳ. After statistical test, there were no statistically significant differences in age and skin type between the two groups. After half a year of follow-up, the average number of treatment times in the experimental group was 5(5, 6) times, lower than that in the control group 7(6, 8) times( T=-9.279, P<0.001). The three and five depilation rates of the experimental group were[70.0%(68.0%, 74.8%), 86.0%(83.0%, 88.0%)], and the final depilation rates of the experimental group were 89.0%(86.7%, 90.0%), all higher than those in the control group[40.0%(37.0%, 42.0%), 72.0%(70.0%, 74.0%), 86.5%(85.0%, 88.0%)]. The differences between the two groups were statistically significant ( T3=43.986, T5=25.151, Tfinal=5.547, all P< 0.001). The intermittent times of single sliding treatment in the experimental group ≤2 times, compared with the control group > 2 times, the comfort level was improved. No blisters or coloration occurred in any of the patients. Patients in the 2 groups were followed up for half a year after the last hair removal. In the 2 groups, there were sparse new fine hairs and soft hair growth that were not included in the hair count, and the included hairs were thinner than those before the treatment. Conclusions:Compared with the control group, the treatment cycle is shorter, the efficiency is higher and the comfort level of the patients is higher in the experimental group.

Objective To compare the differences and clinical value of prognostic evaluation between American Joint Committee on Cancer (AJCC) TNM staging system 7th edition and 8th edition for gastric cancer (GC).Methods The retrospective case-control study was conducted.The clinicopathological data of 1 383 GC patients who were admitted to the First People's Hospital of Changzhou between January 2008 and August 2012 were collected.Distal gastrectomy,proximal gastrectomy + pyloroplasty or total gastrectomy were performed according to preoperative evaluation and intraoperative exploration.Observation indicators:(1) surgical and postoperative situations;(2) follow-up and survival situations;(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition;(4) N staging comparison of AJCC TNM staging system 8th edition;(5) prognostic analysis in N staging of AJCC TNM staging system 8th edition;(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition;(7) prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to October 2017.Measurement data with normal distribution were represented as x ± s.Measurement data with skewed distribution were described as M (range).The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Results (1) Surgical and postoperative situations:1 383 GC patients underwent successful radical gastrectomy,including 923 with distal gastrectomy,165 with proximal gastrectomy and 295 with total gastrectomy.Of 1 383 patients,115 with postoperative complications were improved by symptomatic treatment,including 87 with surgical complications and 28 with non-surgical complications.Postoperative pathological examinations:total number of intraoperative lymph node dissection and number of lymph node metastasis were 25± 12 and 7±4;577 didn't have lymph node metastasis and 806 had regional lymph node metastasis;308 were in early GC and 1 075 in advanced GC.(2) Follow-up and survival situations:1 383 patients were followed up for 1-117 months,with a median time of 34 months.The 1-,3-and 5-year survival rates of 1 383 patients were respectively 90.5％,71.9％ and 61.1％.(3) T staging comparison between AJCC TNM staging system 7th edition and 8th edition:T staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.T staging of 1 383 patients:308,192,65,628 and 190 were respectively detected in T1,T2,T3,T4a and T4b stagings.(4) N staging comparison between AJCC TNM staging system 7th edition and 8th edition:N staging definition between AJCC TNM staging system 7th edition and 8th edition was identical.N staging of 1 383 patients:577,255,207,230 and 114 were respectively detected in N0,N1,N2,N3a and N3b stagings.N3a and N3b were classified as N3 staging of AJCC TNM staging system 7thedition,but they were classified as independent staging of AJCC TNM staging system 8th edition.(5) Prognostic analysis in N staging of AJCC TNM staging system 8th edition:5-year survival rate of patients in N0,N1,N2,N3a and N3b stagings was respectively 85.6％,76.5％,59.4％,45.2％ and 32.5％ based on AJCC TNM staging system 8th edition,with a statistically significant difference in survival (x2 =394.400,P＜0.05).There was a statistically significant difference between N0 and N 1 stagings (x2 =45.630,P＜0.05),between N 1 and N2 stagings (x2 =19.470,P＜0.05),between N2 and N3a stagings (x2 =7.602,P＜0.05) and between N3a and N3b stagings (x2=13.020,P＜0.05).(6) TNM staging comparison between AJCC TNM staging system 7th edition and 8th edition:TNM staging of 366 patients had changes,including 2 in T1N3b staging,2 in T2N3b staging,18 in T3N3b staging,120 in T4aN2 staging,149 in T4aN3a staging,34 in T4bN0 staging and 41 in T4bN2 staging;364 were detected in staging Ⅲ in 7th edition and 8th edition,and sub-staging of staging Ⅲ had a change;2 in T1N3b of ⅡB staging were redistricted into Ⅲ B staging based on AJCC TNM staging system 8th edition.(7) Prognostic analysis in different TNM staging between AJCC TNM staging system 7th edition and 8th edition:according to 7th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in ⅡA staging and 207,70.5％ in ⅡB staging and 136,61.0％ in ⅢA staging and 236,37.5％ in Ⅲ B staging and 333,35.4％ in Ⅲ C staging,with a statistically significant difference in survival among sub-stagings (x2 =228.800,P＜0.05).There was a statistically significant difference in survival among Ⅰ,Ⅱ and Ⅲ stagings (x2=189.000,P＜0.05) and between ⅢA and ⅢB or ⅢC stagings (x2=22.710,18.010,P＜0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2=0.179,P＞0.05),between Ⅱ A and Ⅱ B stagings (x2 =0.265,P＞0.05),and between Ⅲ B and Ⅲ C stagings (x2 =1.550,P＞0.05).According to 8th edition,cases and 5-year survival rate were respectively 247,89.5％ in Ⅰ A staging and 147,83.7％ in Ⅰ B staging and 77,75.9％ in Ⅱ A staging and 205,70.7％ in Ⅱ B staging and 288,53.8％ in ⅢA staging and 258,37.3％ in ⅢB staging and 161,28.5％ in ⅢC staging,with a statistically significant difference in survival among sub-stagings (x2=234.900,P ＜ 0.05).There was no statistically significant difference in survival between Ⅰ A and Ⅰ B stagings (x2 =0.179,P＞0.05) and between Ⅱ A and ⅡB stagings (x2 =0.564,P＞0.05).There was statistically significant differences in survival between Ⅲ A and Ⅲ B or ⅢC stagings (x2 =29.790,43.060,P＜0.05) and between Ⅲ B and Ⅲ C stagings (x2 =7.494,P＜0.05).Further analysis showed that changes of TNM staging system between 7th edition and 8th edition were in T3N3b,T4aN2,T4aN3a,T4bN0 and T4bN2 stagings,5-year survival rate in above stagings was respectively 16.7％,35.8％,30.2％,47.1％ and 26.8％,with statistically significant differences in survival between T3N3b and T4aN2,T4aN3a,T4bN0 and T4bN2 stagings (x2 =19.590,8.039,12.070,3.853,P＜0.05),between T4aN2 and T4aN3a,T4bN2 stagings (x2 =6.529,3.859,P ＜ 0.05),between T4aN3a and T4bN0 stagings (x2 =10.400,P＜0.05) and between T4bN0 and T4bN2 stagings (x2=4.636,P＜0.05).There was no statistically significant difference in survival between T4aN2 and T4bN0 stagings (x2 =3.607,P＞0.05) and between T4aN3a and T4bN2 stagings (x2 =0.029,P＞0.05).Conclusions Compared with AJCC TNM staging system 7th edition,N3a and N3b stagings are classified as independent staging in AJCC TNM staging system 8th edition,and 8th edition is more accurate in prognostic evaluation of GC patients in stage Ⅲ.

Objective To investigate the factors affecting the efficacy of leflunomide combined with medium/low dose corticosteroids in the treatment of progressive IgA nephropathy (IgAN).Methods Clinical and pathological parameters were collected retrospectively in patients of primary IgAN with proteinuria＞ 1.0 g/24 h and chronic kidney disease (CKD) stage 1-3 treated with leflunomide combined with medium/low dose corticosteroids in Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University from Jan 2005 to Dec 2010.According to the treatment effects,patients were divided into complete remission group and non-complete remission group.The biochemical and pathological indexes of the two groups were compared.Results A total of 42 patients were included.The remission rates at 3,6,9 and 12 months were 62％,64％,67％ and 74％,respectively.Seventeen (40.5％) and fourteen (33.3％) patients achieved complete and partial remission after one-year treatment,and the remission rate remained stable within one year after withdrawal of drugs.The 24hour proteinuria was 1.50 (0.67,2.66) g,which was significantly reduced compared with the baseline 2.44 (1.36,3.74) g (P ＜ 0.01).The decrease rate was 31.3％.There was a slight decrease in proteinuriawithin one year after withdrawal of drugs.Estimated glomerular filtration rate (eGFR) remained stable during the treatment and a year of follow-up.No serious adverse event was observed during the followup period.Among 31 responder patients,6(19.4％) patients relapsed.Logistic multivariate regression analysis suggested that the degree of renal interstitial inflammatory infiltration was an independent predictor of complete remission with one-year treatment of leflunomide combined with medium / low dose corticosteroids (HR=0.067,95％ CI 0.008-0.535,P=0.011).Conclusions IgAN treated with leflunomide and medium/low dose corticosteroids can achieve remission in early stage,and the remission rate remains stable after withdrawal of drugs.It is a safe option for the treatment of IgAN.Renal interstitial inflammatory infiltration is an independent predictor of complete remission.

Objective To explore the value of low-dose multi-slice spiral computed tomography (MSCT) multi-stage scan and three-dimensional reconstruction in diagnosis of patients with gastric carcinoma.Methods A total of 40 patients with gastric carcinoma were selected.All the patients were conducted with low-dose MSCT scan,dynamic contrast scanning and three-dimensional reconstruction scan,and then were processed by multi-planar reconstruction,volume reconstruction and CT virtual endoscopy.Primary lesions in gastric cancer,peripheral infiltration and distant metastasis were observed.Results All the patients were satisfied with the low dose MSCT images,and all of them found primary gastric lesions and had satisfactory diagnosis rates of gastric peripheral infiltration,lymph node metastasis and distant metastasis.Conclusion Low-dose MSCT multi-stage scan and three-dimensional reconstruction can reduce the radiation dose and reveal the primary gastric lesions,gastric peripheral infiltration and distant metastasis.

Objective To explore the value of low-dose multi-slice spiral computed tomography (MSCT) multi-stage scan and three-dimensional reconstruction in diagnosis of patients with gastric carcinoma.Methods A total of 40 patients with gastric carcinoma were selected.All the patients were conducted with low-dose MSCT scan,dynamic contrast scanning and three-dimensional reconstruction scan,and then were processed by multi-planar reconstruction,volume reconstruction and CT virtual endoscopy.Primary lesions in gastric cancer,peripheral infiltration and distant metastasis were observed.Results All the patients were satisfied with the low dose MSCT images,and all of them found primary gastric lesions and had satisfactory diagnosis rates of gastric peripheral infiltration,lymph node metastasis and distant metastasis.Conclusion Low-dose MSCT multi-stage scan and three-dimensional reconstruction can reduce the radiation dose and reveal the primary gastric lesions,gastric peripheral infiltration and distant metastasis.