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.

Objective:To analyse the clinical significance of selective single embryo transfer by time-lapse mo-nitoring(TLM)or conventional morphology assessment(CMA)in vitro fertilization/intracytoplasmic sperm in-jection and embryo transfer(IVF/ICSI-ET),and to initially explore the predictive value of Raman spectral analy-sis of embryo culture medium for clinical pregnancy rate.Methods:The study is a prospective randomized con-trolled clinical trial.We assigned 139 patients treated with IVF/ICSI-ET in Reproductive and Genetics Center of Suzhou Municipal Hospital from April 2019 to July 2020,which were randomly assigned to either the CMA or the TLM group.We performed selective single-embryo transfer(fresh cycle and FET)after selecting the optimal em-bryos with TLM or CMA respectively.If the patient's first embryo transfer was unsuccessful,a second one would be performed to compare the differences in the cumulative live birth rate of embryo transfer and other pregnancy outcomes between the two groups.Meanwhile,we collected 15 μl of embryo culture medium at day 3 after IVF/ISCI fertilization for Raman spectroscopy analysis.Results:There were no differences in cumulative live birth,cu-mulative clinical pregnancy,cumulative premature birth,cumulative early spontaneous abortion,cumulative ectopic pregnancy and LGA or SGA between TLM and CMA groups(P＞0.05).The Neonatal sex ratio in the TLM group was lower than that in the CMA group,but the difference was not significant(P＞0.05).Raman spectros-copy analysis of embryo culture medium predicted the clinical pregnancy rate with 67.21％accuracy.Conclu-sions:In young women with a good ovarian reserve,the advantage of using TLM to evaluate embryos is not obvi-ous,so we should remain vigilant that embryo selection based on morphokinetic parameters may affect the sex ratio.Raman spectroscopic analysis of embryo culture medium is not yet able to effectively predict the planting ability of embryos.

Objective:To explore the effects of human chorionic gonadotropin (hCG), progesterone as well as the change of estradiol 12 h after hCG trigger on the outcomes of in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective study was conducted at the Center for Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University. A total of 2 506 patients received IVF/ICSI-ET treatment with gonadotropin-releasing hormone agonist (GnRH-a) protocol from March 2015 to June 2020 were selected. With Spearman rank correlation analysis and path analysis, we explore the relationship among the changes of these hormones and the baseline characteristic of patients, as well as the relationship among the changes of these hormones and the outcomes of IVF treatment.Results:The increase of hCG was accompanied by the rise of progesterone and the decline of estradiol change rate ( r=0.094, P<0.001; r=-0.093, P<0.001). Meanwhile the rise of progesterone was accompanied by the decline of estradiol change rate ( r=-0.089, P<0.001). The dosage of hCG trigger was directly positively correlated to hCG level after hCG trigger, path coefficients (PC) was 0.307 ( P<0.001). Body mass index (BMI) was directly negatively correlated to hCG level and progesterone level after hCG trigger (PC=-0.434, P<0.001; PC=-0.154, P<0.001), whereas positively correlated to estradiol change rate (PC=0.097, P<0.001). Meanwhile the duration and dosage of gonadotropin (Gn) used were positively correlated to progesterone level after hCG trigger (PC=0.102, P<0.001; PC=0.080, P=0.030). hCG level and progesterone level after hCG trigger had positive correlation to oocyte retrieved rate (PC=0.098, P<0.001; PC=0.080, P<0.001). While estradiol change rate was not correlated to oocyte retrieved rate ( P>0.05). Progesterone level on hCG trigger day negatively related to normal fertilization rate (PC=-0.050, P=0.039). hCG level, progesterone level and estradiol change rate after hCG trigger had no correlation with high-quality embryo rate, clinical pregnancy rate and live birth rate (all P>0.05). Conclusion:Oocyte retrieved rate was positively affected by hCG level and progesterone level 12 h after hCG trigger. While normal fertilization rate, high-quality embryo rate, clinical pregnancy rate and live birth rate were not affected by the change of hormones level 12 h after hCG trigger. Therefore we should pay attention to hCG level and progesterone level 12 h after hCG trigger.

Objective:To explore the effects of human chorionic gonadotropin (hCG), progesterone as well as the change of estradiol 12 h after hCG trigger on the outcomes of in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective study was conducted at the Center for Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University. A total of 2 506 patients received IVF/ICSI-ET treatment with gonadotropin-releasing hormone agonist (GnRH-a) protocol from March 2015 to June 2020 were selected. With Spearman rank correlation analysis and path analysis, we explore the relationship among the changes of these hormones and the baseline characteristic of patients, as well as the relationship among the changes of these hormones and the outcomes of IVF treatment.Results:The increase of hCG was accompanied by the rise of progesterone and the decline of estradiol change rate ( r=0.094, P<0.001; r=-0.093, P<0.001). Meanwhile the rise of progesterone was accompanied by the decline of estradiol change rate ( r=-0.089, P<0.001). The dosage of hCG trigger was directly positively correlated to hCG level after hCG trigger, path coefficients (PC) was 0.307 ( P<0.001). Body mass index (BMI) was directly negatively correlated to hCG level and progesterone level after hCG trigger (PC=-0.434, P<0.001; PC=-0.154, P<0.001), whereas positively correlated to estradiol change rate (PC=0.097, P<0.001). Meanwhile the duration and dosage of gonadotropin (Gn) used were positively correlated to progesterone level after hCG trigger (PC=0.102, P<0.001; PC=0.080, P=0.030). hCG level and progesterone level after hCG trigger had positive correlation to oocyte retrieved rate (PC=0.098, P<0.001; PC=0.080, P<0.001). While estradiol change rate was not correlated to oocyte retrieved rate ( P>0.05). Progesterone level on hCG trigger day negatively related to normal fertilization rate (PC=-0.050, P=0.039). hCG level, progesterone level and estradiol change rate after hCG trigger had no correlation with high-quality embryo rate, clinical pregnancy rate and live birth rate (all P>0.05). Conclusion:Oocyte retrieved rate was positively affected by hCG level and progesterone level 12 h after hCG trigger. While normal fertilization rate, high-quality embryo rate, clinical pregnancy rate and live birth rate were not affected by the change of hormones level 12 h after hCG trigger. Therefore we should pay attention to hCG level and progesterone level 12 h after hCG trigger.

Immunoglobulin (IgG) glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases, including systemic lupus erythematosus (SLE), thus underlining the pathogenic role of glycosylation aberration in autoimmunity. This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy. Relative to that in serum samples from the control cohort, IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages (from preconception to the third trimester of pregnancy) and was significantly associated with lupus activity and fetal loss during lupus pregnancy. The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation. The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells (pDCs). RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase (SYK) signaling pathway significantly differed between IgG- and deSia-IgG-treated pDCs. This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG. Finally, the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG. Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.
Humans ; Pregnancy ; Female ; Lupus Erythematosus, Systemic/pathology* ; Signal Transduction ; N-Acetylneuraminic Acid/metabolism* ; Immunoglobulin G ; Dendritic Cells/pathology*

Objective To improve clinically the recognition of fungal infection associated with hemophagocytic lymphohistiocytosis (HLH) in children. Methods Clinical data of 3 children with fungal infection complicated with HLH in our hospital was retrospectively analyzed. Results All the 3 cases complained of recurrent fever, 2 cases with cough and one case with vomiting. Hepatosplenomagaly and lymphadenectasis were found in the medical examination. The time of diagnosis of fungal infection through etiological examination was 5 days after admission. It was further diagnosed as hemophagocytic lymphohistiocytosis after failure of effective antifungal therapy. Routine blood test showed the counts of leukocytes were increased in early stage, while the number of platelets and hemoglobin decreased in different degrees. The recovery is not satisfactory using antifungal therapy alone, and 2 of them are gradually aggravated and treated with mechanical ventilation. On the basis of antifungal therapy, 2 cases were treated under HLH-2004 regimen, 1 received dexamethasone treatment. All the 3 cases received intravenous immune globulin, and showed improvement. Conclusions Fungal infection complicated with HLH in childhood is rare. The effect of simple antifungal therapy on the progression is limited. However, increasing immunosuppressive therapy based on effective antifungal therapy can improve the prognosis.

Aim To discuss the effects and mechanism of Ganoderma lucidum polysaccharides and metformin on myocardial structure and hemodynamics in type 2 diabetic rats.Methods High fat diet combined with intraperitoneal injection of low dose streptozotocin 30 mg· kg -1 was applied to establish rat model of type 2 diabetes mellitus .The diabetic rats were randomly into normal control group ,diabetes group , ganoderma lucid-um polysaccharides group (600 mg· kg -1 ) , metformin group ( 600 mg · kg -1 ) , combination group ( ganoder-ma lucidum polysaccharides 300 mg · kg -1 +metform-in 300 mg· kg -1 ) .After 12 weeks′treatment,the lev-els of fasting serum glucose were determined and the hemodynamic parameters (LVSP,LVEDP,dp/dtmax,-dp/dtmax ) were determined.Collagen volume fraction ( CVF ) was detected by Van Gieson . Immunohisto-chemical method and Western blot were used to detect myocardial tissue MMP-2 protein expression .Results The fasting blood glucose was significantly decreased in the combined treatment group .Combined medication could significantly improve hemodynamic parameters in diabetic rats: reduced LVEP and raised LVEDP , dp/dtmax and -dp/dtmax .CVF was significantly decreased in combination group .The expression of MMP-2 in my-ocardial tissue was significantly inhibited .Conclusions The combination of Ganoderma lucidum polysaccha-ride and metformin can significantly improve the hemo-dynamic parameters in type 2 diabetic rats, and have a preventive effect on diabetic cardiomyopathy . The mechanism may be related to the down regulation of the expression of MMP-2.

Objective To study the effect of ganoderma lucidum polysaccharides combined with metformin on oxidative stress of type 2 diabetic rats. Methods SD rats were fed with high fat diet for 4 weeks and injected with streptozotocin (30 mg·kg-1 ) to produce type 2 diabetic model. The diabetic rats were randomly divided into diabetes model group, ganoderma lucidum polysaccharides group (600 mg·kg-1 ), metformin group (600 mg·kg-1 ), combination group (ganoderma lucidum polysaccharides 300 mg·kg-1+ metformin 300 mg·kg-1 ), After 12 weeks of treatment, the level of fasting blood glucose was determined, and the activity of superoxide dismutase ( SOD), malondialdehyde ( MDA), catalase ( CAT), glutathione peroxidase (GSH-Px), total cholesterol (TC) and triglyceride (TG) were detected. Results The levels of fasting blood glucose in the treatment groups were significantly lower than that in the diabetes model group (P<0. 01). Furthermore, fasting blood glucose in the combination group was significantly lower than that in ganoderma lucidum polysaccharides group and metformin group (P<0. 01). Compared with diabetes model group, serum TC and TG in the treatment groups were significantly lower (P<0. 05, P<0. 01). Serum TC and TG were significantly lower in the combination group than in ganoderma lucidum polysaccharides group and metformin group (P<0. 05, P<0. 01). Compared with diabetes model group, serum SOD levels in the treatment groups were significantly higher (P<0. 01). Compared with ganoderma lucidum polysaccharides group and metformin group, serum SOD levels in the combination group was significantly higher (P<0. 05). Compared with diabetes group, serum MDA levels in the treatment groups were significantly lower (P<0. 01). Serum MDA in the combination group was significantly lower than that in ganoderma lucidum polysaccharides group and metformin group ( P<0. 05). Compared with diabetes model group, serum CAT and GSH-Px in the treatment groups were significantly higher (P<0. 05, P<0. 01). Serum CAT and GSH-Px in the combination group were significantly higher than those in ganoderma lucidum polysaccharides group and metformin group (P<0. 05). Conclusion Ganoderma lucidum polysaccharides combined with metformin could effectively inhibit oxidantion stress in type 2 diabetic rats. The effect was better than ganoderma lucidum polysaccharides or metformin used alone. The possible mechanism may be related to increased activity of SOD, CAT, GSH-Px in vivo and regulation of dyslipidemia.

Aim To study the effects of ganoderma lu-cidum polysaccharides and metformin on myocardial fi-brosis of type 2 diabetic rats and its mechanism. Methods SD rats were fed with high fat diet for 4 weeks, and then were injected with streptozotocin (30mg·kg-1 ) to replicate type 2 diabetic model. The diabetic rats were randomized into normal control group,diabetes group, ganoderma lucidum polysaccha-rides group ( 600 mg · kg-1 ) , metformin group ( 600 mg·kg-1 ) , and combination group( ganoderma lucid-um polysaccharides 300 mg·kg-1 +metformin 300 mg ·kg-1 ) . After 12 weeks’ treatment,the levels of fast-ing serum glucose were determined and the extent of myocardial fibrosis was observed by Picro-sirius red staining. The contents of AGEs in serum were deter-mined by fluorescence spectrophotometer. The activities of CAT and GSH-Px in myocardium were detected. Im-munohistochemical method and Western blot were used to detect myocardial tissue AGEs and CTGF protein ex-pression. Results Combination group could repress patho-proceeding of myocardial fibrosis efficiently, im-prove the activity of CAT and GSH-Px in myocardium and lower the concentration of AGEs in serum, as well as reduce the expression of AGEs and CTGF in myo-cardium. Conclusions Ganoderma lucidum polysac-charides and metformin could prevent myocardial fibro-sis. The possible mechanism may be related to repress-ing oxidative stress of myocardium, lowering serum AGEs and down regulating AGEs and CTGF of myocar-dium.