ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss （RPL） by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus （Shoutai Wan， Juyuan Jian）-of traditional Chinese medicine （TCM） at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c （blank group） and DBA/2 （modeling group） mice separately. Pregnant mice in the modeling group were randomly grouped as follows： high/low-dose Shoutai Wan， high/low-dose Juyuan Jian， model （RPL）， and positive control （dydrogesterone）， with 10 mice in each group. Starting from the day after the detection of the vaginal plug， mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention， the following indicators were measured. ① Macroscopic evaluation： general conditions， uterine wet weight， embryo loss rate， four coagulation parameters ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， fibrinogen （FIB）， and thrombin time （TT）］， and peripheral blood estradiol （E₂） and progesterone （Pg） levels. The decidua with embryos was stained with hematoxylin-eosin （HE） and evaluated by transmission electron microscopy （TEM）. The expression of B-cell lymphoma-2 （Bcl-2）， vascular endothelial growth factor （VEGF）， angiotensin Ⅱ （AngⅡ）， matrix metalloproteinase-2 （MMP-2）， interleukin-6 （IL-6）， leukemia inhibitory factor （LIF）， CXC chemokine ligand 12 （CXCL12）， and microtubule-associated protein 1 light chain 3 homolog （LC3）Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level： The decidua with embryos from the blank， model， high-dose Shoutai Wan， and high-dose Juyuan Jian groups （6 mice per group， with 3 single-cell samples per group， totaling 24 mice） were analyzed by the BD Rhapsody™ platform， and the whole-cell atlas was drawn by uniform manifold approximation and projection （UMAP） dimensionality reduction clustering combined with the single-cell mouse cell atlas （scMCA）. The differentially expressed genes （DEGs） and cell interaction networks were analyzed via Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and CellChat， and the protein-protein interaction （PPI） map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells （natural killer cells， NK cells） from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group， the RPL group showed an increase in the embryo loss rate （P<0.01）， down-regulated expression of Bcl-2， LIF， MMP-2， and Vegf in the decidua with embryos （P<0.05）， up-regulated protein levels of CXCL-12， AngⅡ， and IL-6 （P<0.05）， blocked angiogenesis， apoptosis-inflammation imbalance， and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance， and Shoutai pill showed superior performance in restoring the E₂ level to the Pg level （P<0.05）. ② Single-cell transcriptome analysis indicated that compared with the blank group， the RPL group showed differences in multiple key cell populations such as decidual cells， trophoblast cells， endothelial cells， erythroblasts， NK cells， and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group， high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer （upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes） and macrophages （upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes） through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins， cell adhesion， and programmed cell death， thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells （up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes） and macrophages （up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes）， which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D， CDH5， GDF， and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 （STAT3） and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis （TWEAK） signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7， a decreased proportion of reparative dNK4， and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1， thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method （Shoutai Wan） has an advantage in regulating the phenotypes of unfolded protein， cell adhesion， and programmed cell death， and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method （Juyuan Jian） has an advantage in regulating epithelial-mesenchymal transition （EMT）， angiogenesis， and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.

Objective: To understand the proper handwashing behavior of preschool children and primary school students in Beijing, and to analyze associated family factors to provide reference for further health intervention related to handwashing. Methods: From November to December 2020, parents of 36 kindergartens and 18 primary schools in 9 districts of Beijing were investigated online by using a self designed questionnaire with questionnaire star software. The contents of the survey included the basic situation of children and their families, parents correct knowledge of the prevention of novel coronavirus pneumonia, their perception of the epidemic risk, the provision of handwashing guidance for children, and children s handwashing behavior. Results: The proportion of proper handwashing of preschool children was 70.2%, which was higher than that of primary school students (61.9%) ( χ 2=57.63, P <0.01). The proportion of parents of preschool children who correctly knew handwashing related knowledge (36.2%, 33.4%), had low perception of epidemic risk (28.9%, 25.4%), and provided handwashing guidance (99.1%, 97.9%) was higher than that of parents of primary school students, and the differences were statistically significant ( χ 2=6.72, 22.84, 18.68, P <0.05). But the proportion of parents of preschool children who had high self efficacy (75.7%, 78.2%)was lower compared to parents of primary school students( χ 2=6.43, P =0.04). Multivariate regression results showed that whether preschool children or primary school students, urban areas and parents had high self efficacy, low risk perception, and provided hand washing guidance for children, children were more likely to wash their hands correctly. For preschool children, non only children were 0.79(95% CI =0.69-0.92) times more likely to wash their hands correctly than only children. For primary school students, girls were 1.21(95% CI =1.06- 1.39 ) times more likely to wash their hands correctly than boys, and parents who know knowledge correctly were 1.20(95% CI = 1.04 -1.40) times more likely to know it incorrectly( P <0.05). Conclusion Proper hand washing behavior of preschool children is higher than that of primary school students. Parental awareness of COVID-19 epidemic, handwashing behavior, self efficacy and guidance behavior have effects on the development of children s health behavior. Measures should be taken to enhance parents awareness of infectious diseases and their ability and self efficacy of guiding children in disease prevention.

This article was aimed to study the effect ofQiwei granules on the podocyte in KK-Ay mice kidney. The 28 8-week-old male KK-Ay mice were randomly divided into the model group, low-dosage, middle-dosage and high-dosageQiwei granule group. Eight C57BL/6J mice were used as the normal control. The general conditions, blood glucose and 24 h albuminuria were recorded in the experiment. After 10-week treatment, renal indexes including serum creatinine and urea nitrogen were measured. The kidneys of mice were collected and measured. The hematoxylin and eosin (HE), Masson’s Trichrome, and periodic acid-Schiff (PAS) were used on renal tissues of mice. The immunohistochemical staining for WT-1 was made. Software analysis was combined in the calculation of renal podocyte amount. Western blot was used in the detection of nephrin protein expressions in the kidney of mice. RT-PCR was used in the detection of nephrin mRNA expression. The results showed that compared with the model group, the body weight, blood glucose, 24 h albuminuria and the serum creatinine were obviously decreased after 10-week treatment ofQiwei granules. It can effectively improve the glomerular mesangial proliferation and preserve the podocyte number. Meanwhile, after the treatment ofQiwei granules, the nephrin protein expression and mRNA expression were obviously higher than the model group. It was concluded thatQiwei granules probably managed nephrin expression to improve the podocyte injury in the diabetic nephropathy of KK-Ay mice.

Recently, drug delivery materials have become the hotspot of medical study. Suitable delivery material plays an important role in constructing an excellent drug delivery system. Silk fibroin is a naturally occurring protein polymer with excellent biocompatibility, remarkable mechanical properties, biodegradability and outstanding processability. Due to its unique properties, silk fibroin has become a favorable carrier material for the incorporation and delivery of a range of therapeutic agents. Based on the structure and characteristics of silk fibroin, this article provides an overview of the recent research progress of silk fibroin used as drug delivery materials.
Biocompatible Materials ; Drug Delivery Systems ; Fibroins ; chemistry

Objective To explore the role of sphingosine-1-phosphate receptor (S1PR2) in human coronary artery endothelial cell proliferation in vitro. Methods MTT assay was used to detect cell proliferation in human coronary artery endothelial after treatment of S1P and S1PR2 antagonist JTE-013. Phosphor-ERK and total- ERK level were measured by western blot in endothelial after treatment of S1P and JTE-013. Results 1 μmol/L S1P significantly increased endothelial cells proliferation. S1PR2 antagonist JTE-013 inhibited S1P-induced endothelial cell proliferation in dose-dependent manner. S1PR2 antagonist JTE-013 significantly inhibited S1P-induced phosphor-ERK level in endothelial cells. Conclusion S1PR2 may involve in S1P-induced endothelial cell proliferation through activation of ERK pathway.

This study was aimed to observe the effects on glucolipid metabolism of aqueous extract of traditional South African herb Sutherlandia. Intraperitoneal injection of streptozotocin (STZ) combined with high fat feed method was used in the establishment of type 2 diabetes rat model. Then, rats were randomly divided into the normal group, model group, pioglitazone group, and the Sutherlandia group. Observation was made on changes of body weight, oral glucose tolerance test (OGTT), blood triglycerides (TG), total cholesterol (TC), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C). Western blot method was used to detect IRS-1 expression of skeletal muscle in rats of each group. The results showed that compared with the normal group, body weight in the model group was decreased, and indicators of OGTT, TG, TC, LDL-C were significantly increased (P< 0.05). Symptoms such as increased drink-ing, eating and urine were obvious; and the IRS-1 expression was obviously decreased (P< 0.05). After treatment, compared with the model group, there was no significant body weight increase in the Sutherlandia group or the pi-oglitazone group. Indicators of blood glucose, TG, TC in the Sutherlandia group and the pioglitazone group were ob-viously decreased with no statistical difference (P< 0.05 or P< 0.01). In the Sutherlandia group and the pioglitazone group, IRS-1 expression of skeletal muscle in rats was obviously increased with no statistical difference (P< 0.05). It was concluded that type 2 diabetes rats induced by intraperitoneal injection of STZ combined with high fat feed method can induce glucolipid metabolism disorders. Traditional South African herb Sutherlandia can obviously reduce the blood sugar level, improve blood lipid metabolism, and improve the level of insulin. Sutherlandia can improve the IRS-1 expression of skeletal muscle in rats, relief the insulin resistance, and lower blood sugar. However, the effect of its exact ways required further in-depth study.

This study was aimed to explore the effect of Tang-Nai-Kang (TNK) on trans-differentiation of renal tubular epithelial cell in KKAy mice in order to discuss the possible mechanism. Fifty 12-week-old male KKAy mice were randomly divided into the model group, valsartan group, TNK high-dose, middle-dose and low-dose group, with 10 rats in each group. Ten C57BL/6J mice were used in the normal group. Rats in the model group and normal group were given 0.9% sodium chloride solution. Rats in other groups were given the corresponding drugs. After 8 weeks of gavage administration, kidneys of all mice were sampled and given Mosson and PAS dyeing. Expression distribution of α-smooth muscle actin (α-SMA) and E-cadherin in kidney tissues were observed under immunohistochemical staining. Expression of transforming growth factor-β1 (TGF-β1) was measured by western blot. The results showed that compared with the normal group, the area of renal fibrosis in the model group was significantly increased (P < 0.01); the expression of α-SMA was stronger; and the expression of E-cadherin was weaker. Compared with the model group, the area of renal fibrosis in the valsartan group, TNK high-dose, middle-dose and low-dose groups were significantly decreased (P< 0.01); the expression of α-SMA was weaker (P< 0.01);and the expression of E-cadherin was obviously increased (P < 0.05). The TGF-β1 expression in the model group was significantly higher than that in the normal group (P < 0.01). Compared with the model group, the TGF-β1 expression in the valsartan group, TNK low-dose, middle-dose and high-dose groups were significantly lowered (P<0.01). And the TGF-β1 expression in the TNK high-dose group was even lower than that in the valsartan group. It was concluded that TNK was able to suppress the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cell, and lessen the renal tubule interstitial fibrosis, in order to protect the kidney.

Objective To understand the status of cognitive dysfunction among new onset constipation after stroke and to explore the relationship between them.Methods With a self-designed questionnaire,the general information of 723 inpatients from 10 hospitals in Guangzhou were collected,including general demographic information,stroke situation,defecation situation and Mini-mental State Examination (MMSE)etc.Results The incidence of new-onset constipation,cognitive dysfunction(PSCD)after stroke was respectively 34.6％ and 62.4％.Among whom 31.0％ in ischemic stroke,44.8％ in hemorrhagic stroke,47.4％ in cerebral hemorrhage and infarction.There was statistical significance between stroke type and constipation,and ischemic stroke had statistical significance compared to other two types.During the acute phase reached 41.6％,the recovery phase 31.5％ and the sequelae phase 22.6％.The rates of constipation patients with cognitive dysfunction were higher than patients without cognitive dysfunction,and the difference in directional force,memory,attention and calculation force,reading and expression was statistically significant respectively,among which expression was the most relevant.The difference in the incidence of constipation among different cognitive dysfunction groups was statistically significant,and they were positively correlated,compared with normal cognitive function,the light,medium and severe cognitive dysfunction was 1.519,2.879,3.064 times respectively.Conclusions Medical staff need to be alerted to the possibility that patients with impared cognitive function may be at greater risk of constipation.The preventive and treatment measures should be emphasized in order to improve patients' cognitive function.

OBJECTIVETo investigate the incidence of constipation after stroke and explore the relationship between post-stroke constipation and brain injury.

METHODSUsing a self-designed questionnaire, we collected the general information of 723 inpatients from 10 hospitals in Guangzhou, including the general demographic information, related factors of brain injury, defecation and previous history.

RESULTSThe total incidence of post-stroke constipation was 34.6% in these patients. The incidence of constipation was 31.0% in ischemic stroke patients, 44.8% in hemorrhagic stroke patients, and 47.4% in patients with cerebral hemorrhage and infarction, showing a significant difference between stroke types (χ(2)=12.369, P=0.002). In terms of stages following stroke, the incidence was 41.6% in the acute phase, 31.5% in the recovery phase, and 22.6% in the sequelae phase. In light of lesion locations, the incidence was significantly higher in patients with basal ganglia involvement than in those without (P<0.001). Logistic multivariate analyses showed that stroke type, post-stroke stage, lesion number and basal ganglia involvement were significantly associated with the incidence of constipation.

CONCLUSIONConstipation often occurs in the acute stage following stroke (especially hemorrhagic stroke), and the incidence is higher in patients with basal ganglia involvement.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Brain Injuries ; epidemiology ; Brain Ischemia ; epidemiology ; Cerebral Hemorrhage ; epidemiology ; Constipation ; complications ; epidemiology ; Female ; Humans ; Intracranial Hemorrhages ; epidemiology ; Male ; Middle Aged ; Risk Factors ; Stroke ; complications ; Surveys and Questionnaires ; Young Adult

Objective To investigate the relationship between tumor invasion and changes of mRNA expression of MMP-2, TIMP-2 and CD147 in human pituitary adenoma. Methods 60 patients with pituitary adenoma were devided into two groups, invasive and non-invasive group, by MRI. The expression level of MMP-2,TIMP-2 and CD147 in the samples of pituitary adenoma was measured by semiquantitative reverse transcriptasepolymerase chain reaction (RT-PCR). Pearson analysis was used to reveal the correlation between the expression of each marker. Results The mRNA expression level of MMP-2 and CD147 was significantly higher in invasive pituitary adenoma group than that in non-invasive group (P ＜ 0. 01 ) while the mRNA expression level of TIMP-2was lower in invasive pituitary adenoma group than that in non-invasive group (P ＜0.01 ). According to Pearson analysis, the mRNA expression of MMP-2 was positively correlated with CD147 in invasive pituitary adenoma (r=0. 69, P ＜ 0. 05 ), and M MP-2 was negatively correlated with TIMP-2 in non-invasive pituitary adenoma (r =-0.68, P ＜ 0.05 ). Conclusions The invasion of human pituitary adenoma are closely related to the low expression level of TIMP-2 as well as the high expression level of MMP-2 and CD147. MMP-2, TIMP-2 and CD147 can be used as indicators of tumor invasion of human pituitary adenoma.