Objective The core mechanism of renal insterstitial fibrosis (RIF) is epithelial-mesenchymal transition.This study aimed to investigate the effect of erythropoietin on high glucose-induced epithelial-mesenchymal transition ( EMT) of normal hu-man kidney proximal tubular epithelial cells (HK-2) and its possible mechanism. Mothods HK-2 cells cultured in vitro were ran-domly divided into a blank control group , a high glucose induction group , a mannitol induction group , an EPO induction group , an EPO (5, 10, and 20U/mL) inhibition group, and an Rho kinase inhibitor group.After 24 hours of intervention, the mRNA levels of RhoA and ROCK were determined by RT-PCR, those of E-cadherin and α-smooth muscle actin (α-SMA) proteins detected by immu-nofluorescence staining , and the expression of FN proteins in the supernatant measured by ELISA . Results Compared with the blank control group , the expressions of RhoA and ROCK 1 mRNA were significantly increased in the high glucose induction group (0.945 ±0.132 vs 1.400 ±0.022, 1.007 ±0.002 vs 1.913 ±0.011, P<0.05), but markedly decreased in the 5, 10, and 20U/mL EPO inhibition groups (1.400 ±0.022 vs 1.278 ±0.006, 1.400 ±0.022 vs 0.770 ±0.005, 1.400 ±0.022 vs 0.334 ±0.009, P<0.006) in comparison with the high glucose induction group , and the effects were related to the concentration of EPO .Compared with the blank control, the expression of E-cadherin protein was increased in the high glucose induction group (0.644 ±0.006 vs 0.107 ± 0.004, P<0.05), but remarkably decreased in the 5, 10, and 20 U/mL EPO inhibition groups (0.236 ±0.006, 0.433 ±0.010, 0.521 ±0.010) in comparison with the high glucose induction group (P<0.05), and the effects were also related to the concentration of EPO.Pearson correlation analysis showed a positive correlation between the mRNA expressions of RhoA and ROCK 1 in the high glu-cose induction and EPO inhibition groups . Conclusion EPO can inhibit high glucose-induced epithelial-mesenchymal transition of normal human kidney HK-2 cells and thus delay renal fibrosis , which mignt be related to the RhoA/ROCK signaling pathway .

Objective:To explore the value of improved sequential organ failure assessment (ISOFA) in predicting the prognosis of Klebsiella pneumoniae sepsis. Methods:The clinical data of 379 patients with Klebsiella pneumoniae sepsis admitted to the First Hospital of Jilin University from January 1, 2018 to June 30, 2019 were retrospectively analyzed.They were divided into survival group and death group according to the 28-day prognosis, and the age, gender, C-reactive protein, procalcitonin, grade of bacterial resistance, and the occurrence of septic shock of the two groups were compared.Statistical analysis was conducted by using independent sample t test, Mann-Whitney U test and chi-square test. Risk factors for the prognosis of the disease was analyzed by logistic regression analysis.The correlation between ISOFA and other scoring system including Charlson′s weighted index of comorbidities (WIC), national early warning system (NEWS), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), simplified acute physiology scoreⅡ (SAPSⅡ), mortality in emergency department sepsis score (MEDS), sequential organ failure assessment (SOFA), acute gastrointestinal injury (AGI) was determined by Spearman correlation coefficient.The area under the receiver operating characteristic curve (AUROC) was used to compare the evaluation value of each scoring system for the prognosis of Klebsiella pneumoniae sepsis. The clinical significance of ISOFA risk stratification was evaluated by Kaplan-Meier survival curve. Results:Among the 379 patients with Klebsiella pneumoniae sepsis, 278 were in the survival group and 101 were in the death group.The differences of age, gender, the occurrence of septic shock, grade of bacterial resistance, C-reactive protein, procalcitonin, WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS, SOFA, AGI and ISOFA score between the two groups were all statistically significant ( t=-3.218; χ2=6.781, 24.374 and 27.208, respectively; Z=-3.689, -5.022, -4.396, -4.697, -7.438, -6.348, -6.358, -8.676, -6.680 and -11.658, respectively; all P<0.01). The calculation method of ISOFA was obtained by single factor analysis: ISOFA=SOFA+ 1.5×AGI.Multivariate logistic regression suggested that ISOFA, gender, SOFA, procalcitonin, C-reactive protein, and bacterial resistance were independent risk factors for the prognosis of the disease. The Spearman correlation coefficients of ISOFA and WIC, NEWS, APACHEⅡ, SAPSⅡ, MEDS and SOFA were 0.327, 0.371, 0.614, 0.564, 0.578 and 0.847, respectively. The AUROC and its 95% confidence interval for WIC, NEWS, MEDS, SAPSⅡ, APACHEⅡ, SOFA alone, SOFA plus AGI, and ISOFA to predict the prognosis of Klebsiella pneumoniae sepsis were 0.646 (0.584-0.708), 0.657 (0.597-0.716), 0.712 (0.654-0.771), 0.713 (0.653-0.773), 0.749 (0.693-0.806), 0.788 (0.737-0.838), 0.872 (0.826-0.917) and 0.891 (0.845-0.937), respectively.The results showed that ISOFA had the best predictive effect.The Kaplan-Meier survival curve suggested that there were statistical differences in survival rate among ISOFA low-risk level, medium-risk level, and high-risk level (all P<0.01). Conclusion:ISOFA has important clinical significance in predicting the prognosis of Klebsiella pneumoniae sepsis.

Objective To study the effects of erythropoietin (rhEPO) in high glucose induced proliferation and apopto?sis of human kidney proximal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells cultured in vitro were divided into several groups randomly:blank control group, high glucose group, mannitol group, rhEPO control group, different concentrations of rhEPO treatment groups (5, 10, 20 U/mL) and Rho kinase group. The reverse tran?scription polymerase chain reaction (RT-PCR) was used to evaluate the mRNA levels of RhoA and ROCK after 24 hours. Tetrazolium salt method (MTT) was used to determine the cell proliferation. Cell apoptosis was detected by flow cytometry. Results Compared with blank control group the expression levels of RhoA and ROCK1 mRNA were significantly in?creased in high glucose group (P < 0.05). RhoA, ROCK1 mRNA expressions significantly decreased in rhEPO group than those of high glucose group (P<0.05). There was a positive correlation between the expression levels of RhoA mRNA and ROCK1 mRNA in high glucose group and rhEPO group. MTT method showed that rhEPO significantly promoted the prolifer?ation of HK-2 cells (P<0.05). Flow cytometry analysis showed that high glucose induced apoptosis in HK-2 cells, which was significantly inhibited in rhEPO group and Rho kinase group as compared to that of high glucose group in a concentra?tion dependent manner (P<0.05). Conclusion rhEPO can promote HK-2 cell proliferation and inhibit apoptosis, which may be related to RhoA/ROCK signaling pathway.

OBJECTIVETo analysis the effects of water deficit on the transcript level of SOD, APX, DHAR and MDHAR genes in Scutellaria baicalensis.

METHODThree-month-old S. baicalensis was in glasshouse under water deficit stress, and the transcript level of SOD, APX, DHAR and MDHAR genes were analysis utilized semi-quantitative RT-PCR.

RESULTCompared with the control group, a significant decline of the transcriptional level of APX gene was observed at 70 days after water deficit. The transcript level of DHAR gene was reduced at 30 and 50 days after water deficit. And MDHARI gene was significant declined at 50 days.

CONCLUSIONAsA which is an important antioxidant plays a major role in hydrogen peroxide clear system under water deficit, and maybe have an antagonistic effect to the accumulation of baicalein.

Ascorbate Peroxidases ; genetics ; metabolism ; Ascorbic Acid ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Plant ; drug effects ; Hydrogen Peroxide ; metabolism ; NADH, NADPH Oxidoreductases ; genetics ; metabolism ; Oxidoreductases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; enzymology ; genetics ; metabolism ; Superoxide Dismutase ; genetics ; metabolism ; Time Factors ; Water ; metabolism ; pharmacology

Objective To establish a mouse model by transplanting subcutaneously human pancreatic cancer tissue fragments.Methods Surgically resected pancreatic cancer tissue fragments from patients were transplanted into NOD/SCID mice subcutaneously,and then the growth of the tumor and transplanting it into the next generation were observed.The growth rate,HE staining and immunohistochemistry staining of Ki67 and VEGF were compared.Results We have obtained 13 cases ofpancreatic cancer tissues and 6 cases of biopsy specimens.In 5 cases transplantation was successful,in onemouse model passing to fourth generation,in 4 models to second generation.With the increase of generaions,tumor growth accelerated.HE staining showed later passage cells behavior in an identical manner as the primary cells.Immunohistochemistry staining showed that expressions of Ki67 and VEGF are increasing.Conclusions Through transplanting human pancreatic cancer tissue fragments directly,we have constructed mouse model of pancreatic cancer successfully.With the passage of subculture,the malignant degree and invasiveness may increase.

Malignant tumors are major diseases that endanger human health. Due to their complex and variable microenvironment, most anti-tumor drugs cannot precisely reach the focal tissue and be released in a controlled manner. Intelligent responsive nano carriers have become a hot spot in the field of anti-tumor drug delivery systems. As an excellent nano material, mesoporous silica has the advantages of non-toxic, stable, adjustable pore volume and pore diameter, and easy functional modification on the surface. By virtue of its perceptive response to the tumor microenvironment or physiological changes, it can achieve the targeted drug release or controlled drug release of the drug delivery system in the tissue, making it an ideal carrier for intelligent response drug delivery system. In this paper, we review the design strategies and current research status of smart responsive anti-tumor drug delivery systems based on mesoporous silica, in order to provide a reference for the development of anti-tumor drug nanoformulations.

OBJECTIVETo study the effect of Fuzheng Sanjie recipe in regulating tumor-associated macrophages (TAMs) in Lewis lung cancer mice.

METHODEfforts were made to establish the Lewis lung cancer mouse model, weigh tumors and calculate the anti-tumor rate. The immunohistochemical method was used to examine the infiltration degree of CD68 + in tumor tissues in each group. ELISA was used to examine the content of IFN-γ, TGF-β, IL-4, IL-13, IL-6, IL-10, IL-12, TNF-α in mice serum.

RESULTCompared with the tumor-bearing model group, all of the other groups showed higher tumor inhibition rates, i. e. 50.28% for the DDP group, 34.37% for the TCM-preventing group and 66.76% for the Chinese and western medicine group, with statistical difference (P < 0.05), but without statistical difference in the infiltration degree of CD68+. The expressions of the IFN-γ, IL-6, IL-12 in tumor-bearing groups were lower than that in the blank control group, but with higher contents of IL-4, IL-13, TGF-β. Intervened with different drugs, there were significant differences in content among some relevant cytokines (P < 0.05), as well as statistical differences among the TCM prevention group, the Chinese and western medicine group and the tumor-bearing control group (P <0. 05) , but without statistical difference in TNF-α and IL-10 content from the tumor-bearing control group (P < 0.05).

CONCLUSIONFuzheng Sanjie recipe could reverse the immune remodeling effect and control the tumor growth by down-regulating the expressions of IL-4, IL-13, TGF-α in lung cancer immune microenvironment and up-regulating the expression of IFN-γ.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-13 ; blood ; Lung Neoplasms ; blood ; drug therapy ; immunology ; Macrophages ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

Objective To explore the molecular mechanisms involved in hypokalemic salt-losing tubulopathies ( SLTs) through genetic screening of WNK gene in patients with SLTs. Methods Forty-four kindreds of SLTs were diagnosed Batter's syndrome or Gitelman's syndrome after CLCNKB and SLC12A3 sequencing and analysis, 8 of whose phenotype can not be simply attributed to CLCNKB or SLC12A3 mutations. Primers for PCR-amplified exons of WNK4 and WNK1 gene in genomic DNA were designed, and direct sequencing was performed to analyse the PCR products. Results Two missense mutations of WNK1, Ile~(1172)→ Met (I1172M) and Ser~(2047) → Asn (S2047N), were identified. Both of these 2 mutations segregated with the disease in SLTs kindred. Conclusion Two heterozygote missense mutations of WNK1 gene (I1172 M and S2047N) were found in 8 SLTs kindreds, indicating that WNK1 might be another gene responsible for hypokalemic salt-losing tubulopathies.

The active ingredients in traditional Chinese medicine have been reported to possess significant pharmacological activity and played an important role in clinical treatments. However, lots of the active ingredients in traditional Chinese medicine suffer from disadvantages such as low solubility, high melting point and low stability that results in low bioavailability and limit its clinical application. Crystal structure plays an important role in improving physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine. This review concludes the research advances of several crystal forms used in the active ingredients in traditional Chinese medicine in terms of polymorph, cocrystal, amorphous/coamorphous and nanocrystal. And the effects of crystal forms on the physicochemical properties and efficacy of the active ingredients in traditional Chinese medicine were reviewed. This research may be useful for the formulation preparation and development of the active ingredients in traditional Chinese medicine.

To analyze the constituents in supercritical fluid CO2 extraction (SFE-CO2) of Radix caulophylli, the Radix caulophylli was extracted with SFE-CO2, and analyzed by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis with a DB-5MS capillary column (30 mm x 0.32 mm ID, 0.25 microm film thickness) was used. The inlet temperature was maintained at 280 degrees C. The column oven was held at 80 degrees C for 2 min, then programmed from 80 to 280 degrees C at 5 degrees C x min(-1) and, finally, held for 4 min. Helium at a constant flow rate of 2.0 mL x min(-1) was used as the carrier gas. The mass spectrometry conditions were as follows: ionization energy, 70 eV; ion source temperature, 200 degrees C. The mass selective detector was operated in the TIC mode (m/z was from 40 - 500). For the first time 49 peaks were separated and identified, the compounds were quantitatively determined by normalization method, and the identified compounds represent 97.44% of total GC peak areas. Viz, n-hexadecanoic acid (31.4%), (E, E) -9, 12-octadecadienoic acid (26.54%), (Z)-7-tetradecenal (9.4%), hexadecenoic acid (3.23%), 10-undecenal (3.22%), octadecanoic acid (2.25%), and caulophylline (1.76%) etc. The results will provide important foundation for understanding the constituents and further exploitation of Radix caulophylli.
Carbon Dioxide ; Caulophyllum ; chemistry ; Chromatography, Supercritical Fluid ; Gas Chromatography-Mass Spectrometry ; Linoleic Acid ; analysis ; Palmitic Acid ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry