ObjectiveTo evaluate the clinical efficacy and safety of Baoshen prescription in the treatment of stage Ⅳ diabetic nephropathy （DN） in the patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction， and to explore the mechanism of this prescription delaying the disease progression. MethodsA randomized， controlled， double-blind， multicenter clinical trial was conducted， in which 94 stage Ⅳ DN patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction were randomly assigned into Baoshen prescription and control groups （47 cases）. The treatment lasted for 12 weeks. The primary efficacy indicators were mainly renal function indexes， including urine albumin-to-creatinine ratio （UACR）， 24-hour urine total protein （24 h-UTP）， serum creatinine （SCr）， and estimated glomerular filtration rate （eGFR）. The secondary efficacy indicators were metabolic memory of hyperglycemia， podocyte epithelial-to-mesenchymal transdifferentiation-related indexes， and TCM syndrome score. ResultsAfter 12 weeks of treatment， the Baoshen prescription group showed lowered levels of advanced glycation end products （lgAGEs）， connective tissue growth factor （CTGF）， type Ⅳ collagen （Col-Ⅳ）， receptor of AGEs （RAGE）， urinary fibroblast-specific protein-1 （FSP-1）， UACR， 24 h-UTP， and glycated hemoglobin （HbAlc） （P<0.05）， and an upward trend of miR-21 mRNA. The control group showed elevated levels of SCr and UREA and lowered levels of urinary FSP-1， eGFR， and HbAlc （P<0.05）. After treatment， the Baoshen prescription group had lower levels of lgAGEs， CTGF， urinary FSP-1， SCr， UACR， and 24 h-UTP and higher levels of Col-Ⅳ and eGFR than the control group （P<0.05）. In addition， the Baoshen prescription group showed statistically significant differences in SCr， eGFR， UACR， and 24 h-UTP before and after treatment （P<0.05）. ConclusionBaoshen prescription can effectively improve the renal function， reduce the urinary protein level， and alleviate clinical symptoms in stage Ⅳ DN patients with syndromes of Qi-Yin deficiency and kidney collateral stasis and obstruction. The mechanism may be related to the metabolic memory of hyperglycemia and epithelial-to-mesenchymal transdifferentiation of podocytes.

Background Protein tyrosine phosphatase non-receptor type II (PTPN2) is essential for the regulation of inflammation and immunity, but the specific mechanism of action of Ptpn2 in silicosis is unknown. Objective To investigate the regulatory role of overexpression of Ptpn2 in SiO₂-mediated inflammatory response in alveolar type II epithelial cells based on transcriptome sequencing. Methods This study was an in vitro study. A negative control group (vector transferred) and an overexpression of Ptpn2 group of mouse lung epithelial cell line MLE-12 cells were firstly constructed. Transcriptome sequencing was performed to detect differentially expressed genes (DEGs), differentially expressed mRNAs, and differentially expressed ncRNAs in the two groups of MLE-12 cells, and then the DEGs were analyzed by the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Constructed MLE-12 cells and A549 cells were stimulated using SiO₂ suspension, and divided into a negative control group (vector transferred), an overexpression of Ptpn2 group, a negative control + SiO₂ group, and an overexpression of Ptpn2 + SiO₂ group, respectively. Protein expressions of tumor necrosis factor-α (TNF-α) and interleukin (IL)-17A, IL-2, IL-1β were detected by Western blot. Positive TNF-α expression was detected by immunofluorescence staining. Results The results of Western blot showed that the protein expression level of PTPN2 was up-regulated in the overexpressed Ptpn2 group compared with the negative control group (P < 0.05). The volcano plot and clustering heat map showed that there were 1122 DEGs, 3681 differentially expressed mRNAs, and 474 differentially expressed ncRNAs in the overexpressed Ptpn2 group compared with the negative control group. The results of GO enrichment showed that, in terms of the biological process, the DEGs were mainly related to the regulation of metal ion transport, extracellular matrix organization, and extracellular structural organization; in terms of cellular composition, the DEGs were mainly related to collagen-containing extracellular matrix, receptor complexes, and basement membranes; in terms of molecular function, the DEGs were mainly related to the extracellular matrix structural constituents, glycosaminoglycan binding, and semaphorin receptor binding. The results of KEGG enrichment showed that the DEGs were closely related to cytokin-cytokine receptor interaction, IL-17 signaling pathway, TNF signaling pathway, and chemokine signaling pathway. In MLE-12 and A549 cells, the results of Western blot showed that the protein expression levels of TNF-α , IL-17A, IL-2, and IL-1β were up-regulated in the negative control + SiO₂ group compared with the negative control group (P < 0.05), and the protein expression levels of TNF-α, IL-17A, IL-2, and IL-1β were down-regulated in the overexpression of Ptpn2 + SiO₂ group compared with the negative control + SiO₂ group (P < 0.05). The immunofluorescence staining showed that the expression of TNF-α was increased in the negative control + SiO₂ group compared with the negative control group, and decreased in the overexpression of Ptpn2 + SiO₂ group compared with the negative control + SiO₂ group. Conclusion Overexpression of Ptpn2 inhibits SiO₂-mediated inflammatory response in MLE-12 cells and A549 cells.

OBJECTIVE To establish a method for the determination of the concentrations of anti-tumor lead compounds purine benzamides PLB-E and PLB-P in rat plasma by HPLC and apply to study pharmacokinetics. METHODS The established HPLC was used to determine the plasma drug concentrations of rats at different time points after intravenous administration of 5, 10, 20 mg·kg–1 (low, medium, high doses) of PLB-E and PLB-P, and the pharmacokinetic parameters of each compound were calculated using DAS 3.3.0 software. RESULTS PLB-E and PLB-P had good linear relationship in the range of 2–120, 3–60 μg·mL–1, respectively(r2>0.999). The RSD of inter-day and intra-day precision were <15%. The extraction recoveries were 87.48%–92.84% and 88.24%–92.60%, respectively. The main pharmacokinetic parameters of PLB-E and PLB-P after a single intravenous injection of 5, 10, 20 mg·kg–1 were as follows, the average Cmax was (20.30±2.39), (40.63±3.40), (63.62±7.55)mg·L–1 and (13.21±1.40), (24.87±1.33), (32.83±0.65)mg·L–1, respectively. AUC(0-∞) were (104.67±48.39), (177.42±84.11), (194.32±91.48)mg·h·L–1 and (106.75±54.21), (179.90±93.59), (253.56±126.17)mg·h·L–1, respectively. Tmax of each dose was 0.08 h. CONCLUSION The HPLC method established in this study meets the requirements for the determination of biological samples through methodological verification, which is applicable to the determination of the concentration of PLB-E and PLB-P in rat plasma and the pharmacokinetic study. The pharmacokinetic process of PLB-E and PLB-P in rats conforms to the two-compartment model, and conforms to the nonlinear kinetic elimination.

Objective To establish a mouse model of pregnancy pain-depression comorbidity induced by chronic unpredictable stress(CUS),complete Freund's adjuvant(CFA),and formalin,and to systematically evaluate the associated phenotypes and preliminarily explore the pathological basis of the comorbidity.Methods Eight-week-old C57BL/6J female mice were randomly strarified divided into a control group(no intervention before pregnancy)and a CUS model group(CUS intervention before pregnancy)based on sucrose preference test(SPT)data.After completing the CUS treatment,female and male mice were paired and mated.Pain was induced by injecting 50％CFA and 5％formalin in the right hind foot during pregnancy to create a model of pregnancy pain-depression comorbidity.The experiment was divided into 8 subgroups:control-blank group,CUS-blank group,control-CFA group,CUS-CFA group,control-formalin group,CUS-formalin group,control-CFA+formalin group,and CUS-CFA+formalin group,with 10 mice in each group.The mice in each group were subject to behavioral tests,including the SPT,forced swimming test,tail suspension test,and open field test before and after CUS intervention,during pregnancy,and after delivery.Pain sensitivity changes were measured using mechanical allodynia and thermal hyperalgesia tests.Mice were then euthanized.Levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in hippocampus,as well as cortisol and adrenocorticotropic hormone(ACTH)in serum,were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the control-blank group,the CUS-blank group showed a significant depression-like behavior with reduced pain threshold(P＜0.001).The control-CFA+formalin group showed a decrease in pain threshold after both CFA injection and formalin injection(P＜0.01).Compared with the control-blank and control-formalin groups,the pain threshold was significantly lower in the CUS-formalin group(P＜0.01),with a sequential decrease among the three.Compared with the control-blank and control-CFA groups,the pain threshold was significantly lower in the CUS-CFA group(P＜0.001),with a sequential decrease among the three.Compared with the control-blank and control-CFA+formalin groups,the mechanical pain threshold of mice in the CUS-CFA+formalin group was significantly lower(P＜0.001)and the thermal radiation tolerance time was shorter(P＜0.01),both with sequential decreases among the three.Compared with the control-CFA+formalin and the CUS-blank groups,the CUS-CFA+formalin group had a significantly lower percentage of sucrose preference(P＜0.001),longer immobility time during the forced swimming test(P＜0.001)and tail suspension test(P＜0.001),reduced central exploration time in the open field test(P＜0.001),reduced total exploration distance(P＜0.001),and reduced percentage of distance traveled for central exploration(P＜0.001).Compared with the control-CFA+formalin and CUS-blank groups,the serum cortisol and ACTH levels of the CUS-CFA+formalin group were significantly higher(P＜0.01),and the levels of IL-6 and TNF-α in the hippocampus were higher(P＜0.05).Conclusion The combination of CUS+CFA+formalin injections is an ideal method for establishing a C57BL/6J mouse model of pregnancy pain-depression comorbidity.The behavioral changes in model mice may be attributed to the regulation of inflammatory response in hippocampus and hormone levels in the hypothalamic-pituitary-adrenal(HPA)axis.

Objective To investigate the clinical effect of transverse island fasciocutaneous penile flap in the treatment of meatus and navicular fossa stricture.Methods Fifteen patients with urethral reconstruction with transverse island fasciocutaneous penile flap from October 2014 to December 2018 were enrolled.Six patients had a history of urethroscopic surgery,three had a history of lichensclerosus,three had a history of urethral dilation,and three had no obvious causes.All patients underwent transverse incision under the coronal sulcus,and after fully dissecting the urethra,the urethra was opened longitudinally ventrally.After measuring the actual length of stenosis,the irradiance fascia flap with the corresponding length of the incision was reconstructed.The patients were reviewed at 1 and 3 months after operation,and any complications such as recurrence or urinary fistula were recorded.The urine flow rate was tested 3 months after surgery.Results All 15 patients in this group underwent a successfully operation.The actual measurement of urethral stricture length was 0.5-4.0 cm during operation,with the average of 2.82 cm.Three months after the operation,the urine flow rate ranged from 13.5 ml/s to 23.7 ml/s,with an average of 18.5 ml/s.The overall successful rate was 93.3％ (14/15).The rate of post-operative fistula was 20.0％ (3/15).Two cases complained of needle-like fistula at the incision.One case healed after 3 months,and the other gave up further treatment.One patient developed urethral stricture and urethral skin spasm again 1 month later and was surgically repaired again.Conclusions The initial experience of pedicled island fascia flap for the treatment of urethral stenosis and scaphoid stenosis is safe,feasible and effective for the treatment of urethral stricture.

Objective To investigate maternal zinc metabolism and the changes of zinc-related factors as metallothionein-1 (MT1) and zinc transporter-1 (ZnT1) in certain types of congenital heart diseases (CHD).Methods Fifteen infants with interventricular septal defect,12 infants with atrial septal defect and 7 infants with tetralogy of Fallot,together with their mothers were enrolled,and normal infants and their mothers were enrolled by a ratio of 1 ∶ 1 with the above three types of CHD diseases.General conditions of the mothers,along with their diets and zinc-containing drug supplementation during the pregnancy,were surveyed.Maternal blood zinc levels and serum alkaline phosphatase activities at gestation week 32 and delivery or induced abortion,and the protein and mRNA expressions of MT1 and ZnT1 in maternal serum and placental tissue at delivery or induced abortion were assayed.Results The general conditions were comparable between the CHD group and control group.The ratio of the mothers taking more zinc-rich food was significantly lower in the CHD group than in the control group.Circulating zinc levels in interventricular septal defect (73.55±5.79 μmol/L),atrial septal defect (72.66±5.82 μmol/L) and tetralogy of Fallot (68.72±6.72 μmol/L) groups were significantly lower than those in the control groups (82.77± 7.88,84.58 ± 7.55 and 85.66 ± 7.30 μmol/L) at delivery (P all ＜ 0.05).Similar change patterns were seen for serum alkaline phosphatase activities.The relative quantities of serum MT1 and ZnT1 proteins in interventricular septal defect (73.22±36.54 and 68.55± 27.82),atrial septal defect (64.29± 38.26 and 74.55 ± 29.67) and tetralogy of Fallot (67.88± 30.50 and 70.13±29.65) groups were significantly lower than those in their corresponding control groups (166.31±67.43and 97.67±30.22,182.56±71.40 and 111.65±32.70,and 173.81±62.36 and 108.27±28.52,P＜0.01 or P＜0.05).The relative quantities of placental MT1 and ZnT1 proteins and mRNA expressions in interventricular septal defect (protein quantities 0.438±0.096 and 0.384±0.061,mRNA expressions 1.23±0.82 and 0.96±0.39),atrial septal defect (0.427±0.093 and 0.377±0.059,1.17±0.70 and 0.85±0.40) and tetralogy of Fallot (0.414±0.111 and 0.336±0.066,1.31±0.97 and 0.90±0.38) groups were significantly lower than those in their corresponding control groups (protein quantities 0.565±0.083 and 0.541±0.090,mRNA expressions 2.78± 1.06 and 1.67±0.33;protein quantities 0.622±0.136 and 0.493±0.079,mRNA expressions 2.85±0.89 and 1.72±0.38;protein quantities 0.637±0.125 and 0.521±0.089,mRNA expressions 3.21 ± 0.99 and 1.61±0.29;P＜0.01 or P＜0.05).Conclusion Mothers with their fetus of certain types of CHD are found zinc deficiency,and down-regulation of MT1 and ZnT1 expressions in the serum and placenta may involve in the pathogenesis of CHD when maternal zinc deficiency.

Objective: To investigate the effects of maternal exposure to ambient fine particles (PM_2.5) in Fuzhou during pregnancy on immune responses to ovalbumin (OVA) in neonatal rats and the possible mechanisms. Methods: Pregnant Sprague-Dawley rats were randomly assigned into four groups (ten in each): filtered air (FA) plus normal saline (NS), airborne PM_2.5 plus NS (PM_2.5-NS), FA plus OVA (FA-OVA) and PM_2.5 plus OVA (PM_2.5-OVA) groups. Pregnant dams in the PM_2.5 exposure groups were placed in a PM_2.5 exposure chamber in which the PM_2.5 concentration was equal to the ambient air from the beginning of gestation till delivery, whereas the other dams inhaling air without particulate matters were put into a clean chamber. OVA sensitization was conducted through intraperitoneal injection of OVA at 50 μg per dam at 4 and 9 days of gestation, followed by inhalation of atomized 1% OVA for 30 min at 18, 19 and 20 days of gestation. Dams without OVA sensitization were given NS in the same way. Levels of interleukin (IL)-4, IL-5 and interferon-γ (IFN-γ) in neonatal rats' plasma were measured by enzyme-linked immunosorbent assay just after birth. Protein levels of transcription factors GATA-3 and T-bet in lung were analyzed by Western-blotting. Changes in microRNA(miR)-146a and miR-146b in spleen were detected by real-time polymerase chain reaction. Histological changes in lung were observed under light microscope. One-way analysis of variance and LSD test were used as statistical methods. Results: (1) IL-4 level in plasma was significantly increased in PM_2.5-NS [(18.56±7.04) ng/L], FA-OVA [(34.04±7.06) ng/L] and PM_2.5-OVA groups [(45.67±8.18) ng/L] as compared with that in FA-NS group [(10.51±2.88) ng/L], and the level of IL-4 in PM_2.5-OVA group was higher than that in PM_2.5-NS and FA-OVA groups (F=54.667, P<0.001). Significantly increased IL-5 level in plasma was found in FA-OVA and PM_2.5-OVA groups as compared with that in FA-NS group (F=6.253, P=0.023). Among the four groups, FA-OVA group showed significantly increased IFN-γ level in plasma (F=28.604, P<0.001). (2) GATA-3 level in lung tissues was significantly increased in PM_2.5-NS (31.09±3.54), FA-OVA (35.24±5.00) and PM_2.5-OVA groups (47.81±3.63) as compared with that in FA-NS group (24.19±3.12), and higher in PM_2.5-OVA group than in PM_2.5-NS and FA-OVA groups (F=96.581, P<0.001). T-bet level was significantly lower in FA-OVA and PM_2.5-OVA groups than in FA-NS group. Moreover, PM_2.5-OVA group showed decreased T-bet level as compared with that in PM_2.5-NS and FA-OVA groups (F=30.852, P<0.001). (3) Expression of miR-146a in spleen was significantly enhanced in PM_2.5-NS (1.72±0.27), FA-OVA (1.56±0.37) and PM_2.5-OVA groups (3.06±0.52) than in FA-NS group (1.05±0.25). Moreover, PM_2.5-OVA group showed enhanced expression of miR-146a as compared with that in PM_2.5-NS and FA-OVA groups (F=42.276, P<0.001). Changes in the expressions of miR-146b were similar to those in miR-146a (F=28.776, P<0.001). (4) Stenosis or disappearance of alveolar spaces, accompanied with infiltration of inflammatory cells in interstitial substance and congestion in alveolar septum, was seen in FA-OVA and PM_2.5-OVA groups and conditions in the latter group were more severe. Conclusions Intrauterine exposure to ambient PM_2.5 negatively affects fetal lung development and immunological function in rats, especially when the dams are sensitized with OVA during pregnancy.

High incidence of postoperative abdominal adhesions after gynecological laparoscopy, which may induce a line of serious complications such as chronic pelvic soreness, intestinal obstruction, infertility, etc, has been clinically encountered with the major problems unsolved thus far. By further upgrading the merits of gynecological laparoscopy, including the small operative scope, mild injury and fine manipulation, coupled with the application of anti-adhesion therapies, reduced post-operative adhesions and improved clinical outcomes can be expected. Research has been actively persisted on this area in recent years to achieve new progress, especially in the development of anti-adhesion materials, as well as their clinical application and evaluation of the effects, which is overviewed in this review.

Objective To investigate the effect of simulated microgravity on the proliferation and differentiation of the human megakaryocyte cells in vitro. Methods The fourth generation rotating cell culture system (RCCS-4) was used to generate the simulated microgravity environment. The cell viability was assessed by trypan blue staining method. The proliferation of cells was assessed by cell counting method and CCK8 method. The CD41+/CD61+ cells rate and the cells cycle were detected by flow cytometry. The expression levels of thrombopoietin receptor (c-mpl) and transcription factors were detected with RT-PCR. Results After 24, 48, 72 h, culture under simulated microgravity resulted in a significant decrease in the cell number , proliferative activity, cells in the G2/M phase and levels of c-mpl mRNA expression in comparison with that under the normal gravity (P < 0.05). After 48 h and 72 h culture, CD41+/CD61+ cells ratio decreased and RUNX-1 mRNA expression was down-regulated in cells of the group SMG compared with that of the group NG (P < 0.05). Conclusion Microgravity can inhibit the proliferation and differentiation of human megakaryocyte cells in vitro. The mechanism may be that TPO/c-mpl pathway was inhibited by down regulating the expression of c-mpl which transcriptional inhibition lead to.

Objective To investigate the relationship between maternal exposure to airborne inhalable particulate matters (PM10) in the first trimester of pregnancy and the risk of fetal congenital deformity.Methods Relationship between exposure to airborne PM10 during the 1st,2nd,3rd and 1-3 months of gestation and the risk of fetal anomalies in 203 pregnant women with deformed fetuses,which paralleled to normal pregnant women with discrepancy of conception date ＜30 days (control),were retrospectively analyzed by a case-control study from May 14,2007 to April 30,2012 in Fujian Medical University and Fuzhou General Hospital.Multivariate Logistic regression adjusted for potential confounders including maternal age,gravidity and parity was performed for data analysis.Results According to the inclusion and exclusion criteria,178 pregnant women with deformed fetuses and 356 controlled cases were enrolled.The average levels of PM10 exposed in pregnant women Withfetal cardiovascular anomalies during 1-3 months of gestation were significantly higher than in the controls [(73.80±11.55) μg/m3 vs (70.49±10.83) μg/m3] (t=2.066,P=0.040),but PM10 exposure in the 1st,2nd and the 3rd month of pregnancy were comparable between the observed and control groups [(74.00±17.34) μg/m3 vs (71.70±15.39) μg/m3,t=0.992,P=0.322;(75.15±16.80) μg/m3 vs (71.38±15.66) μg/m3,t=1.625,P=0.106; (70.28±15.84) μg/m3 vs (69.41± 15.84) μg/m3,t=0.383,P=0.702].There were no significant differences of PM10 exposure levels when compared between facial-cervical anomalies,neurologic anomalies and total anomalies and their corresponding controls (P＞0.05).Each increased quartile of the PM10 exposure levels was associated with an elevated risk of fetal cardiovascular anomalies by 1.218 folds (OR =2.218,95 ％ CI:1.232-3.994,P=0.008).Paired multivariate Logistic regression analysis revealed,a positive correlation between PM10 levels at 1-3 months of gestation and fetal cardiovascular anomalies (OR =1.106,95％CI:1.035-1.183,P =0.003).Conclusions Maternal exposure to PM10 in the first trimester of pregnancy possibly exerts negative effects on fetal cardiovascular anomalies.