Objective To investigate the protective effect of recombinant Schistosoma japonicum cystatin(rSj-Cys)against acute liver injury induced by lipopolysaccharide(LPS)and D-GalN in mice.Methods Adult male C57BL/6J mice with or without LPS/D-GaIN-induced acute liver injury were given intraperitoneal injections of rSj-Cys or PBS 30 min after modeling(n=18),and serum and liver tissues samples were collected from 8 mice in each group 6 h after modeling.The survival of the remaining 10 mice in each group within 24 h was observed.Serum levels of ALT,AST,TNF-α and IL-6 of the mice were measured,and liver pathologies was observed with HE staining.The hepatic expressions of macrophage marker CD68,Bax,Bcl-2 and endoplasmic reticulum stress(ERS)-related proteins were detected using immunohistochemistry or immunoblotting,and TUNEL staining was used to detect hepatocyte apoptosis.Results The survival rates of PBS-and rSj-Cys-treated mouse models of acute liver injury were 30%and 80%at 12 h and were 10%and 60%at 24 h after modeling,respectively;no death occurred in the two control groups within 24 h.The mouse models showed significantly increased serum levels of AST,ALT,IL-6 and TNF-α and serious liver pathologies with increased hepatic expressions of CD68 and Bax,lowered expression of Bcl-2,increased hepatocyte apoptosis,and up-regulated expressions of ERS-related signaling pathway proteins GRP78,CHOP and NF-κB p-p65.Treatment of the mouse models significantly lowered the levels of AST,ALT,IL-6 and TNF-α,alleviated liver pathologies,reduced hepatic expressions of CD68,Bax,GRP78,CHOP and NF-κB p-p65,and enhanced the expression of Bcl-2.In the normal control mice,rSj-Cys injection did not produce any significant changes in these parameters compared with PBS.Conclusion rSj-Cys alleviates LPS/D-GalN-induced acute liver injury in mice by suppressing ERS,attenuating inflammation and inhibiting hepatocyte apoptosis.

Objective To investigate the protective effect of recombinant Schistosoma japonicum cystatin(rSj-Cys)against acute liver injury induced by lipopolysaccharide(LPS)and D-GalN in mice.Methods Adult male C57BL/6J mice with or without LPS/D-GaIN-induced acute liver injury were given intraperitoneal injections of rSj-Cys or PBS 30 min after modeling(n=18),and serum and liver tissues samples were collected from 8 mice in each group 6 h after modeling.The survival of the remaining 10 mice in each group within 24 h was observed.Serum levels of ALT,AST,TNF-α and IL-6 of the mice were measured,and liver pathologies was observed with HE staining.The hepatic expressions of macrophage marker CD68,Bax,Bcl-2 and endoplasmic reticulum stress(ERS)-related proteins were detected using immunohistochemistry or immunoblotting,and TUNEL staining was used to detect hepatocyte apoptosis.Results The survival rates of PBS-and rSj-Cys-treated mouse models of acute liver injury were 30%and 80%at 12 h and were 10%and 60%at 24 h after modeling,respectively;no death occurred in the two control groups within 24 h.The mouse models showed significantly increased serum levels of AST,ALT,IL-6 and TNF-α and serious liver pathologies with increased hepatic expressions of CD68 and Bax,lowered expression of Bcl-2,increased hepatocyte apoptosis,and up-regulated expressions of ERS-related signaling pathway proteins GRP78,CHOP and NF-κB p-p65.Treatment of the mouse models significantly lowered the levels of AST,ALT,IL-6 and TNF-α,alleviated liver pathologies,reduced hepatic expressions of CD68,Bax,GRP78,CHOP and NF-κB p-p65,and enhanced the expression of Bcl-2.In the normal control mice,rSj-Cys injection did not produce any significant changes in these parameters compared with PBS.Conclusion rSj-Cys alleviates LPS/D-GalN-induced acute liver injury in mice by suppressing ERS,attenuating inflammation and inhibiting hepatocyte apoptosis.

Objective:To evaluate the effectiveness and safety of phacoemulsification cataract extraction combined with intraocular lens implantation (PEI) plus goniosynechilysis (GSL) and goniotomy (GT) for advanced primary angle-closure glaucoma (PACG).Methods:An observational case series study was performed.Fifty eyes of 50 patients with advanced PACG were enrolled in Zhongshan Ophthalmic Center from August 2020 to June 2021.All the patients received PEI+ GSL+ GT and were followed up for over 6 months, with a mean follow-up of 7.5 (6, 10) months.Intraocular pressure (IOP) was measured with a Goldmann applanation tonometer.Best corrected visual acuity (BCVA) was examined with an ETDRS chart and converted to logarithm of the minimum angle of resolution (LogMAR) units for analysis.Types and number of anti-glaucoma medications applied before and after surgery, and the surgical complications were collected.Success rate of surgery was calculated.Complete surgical success was defined as an IOP of 5-18 mmHg (1 mmHg=0.133 kPa) with a reduction of 20% from baseline without anti-glaucoma medication, no vision-threatening complications, no loss of light perception, and no reoperation.Qualified success was defined as an IOP of 5-18 mmHg with a reduction of 20% from baseline with or without anti-glaucoma medication, no vision-threatening complications, no loss of light perception, and no reoperation.This study adhered to the Declaration of Helsinki.This research protocol was approved by an Ethics Committee of Zhongshan Ophthalmic Center (No.2021KYPJ177). Written informed consent was obtained from each subject before entering the cohort.Results:The mean preoperative IOP was (28.81±7.81)mmHg, and the IOP at the end of follow-up was (13.41±4.10)mmHg, showing a statistically significant decrease ( t=12.260, P<0.001). The postoperative IOP was decreased by 13.80 (9.10, 19.40)mmHg, with a percentage decrease of 51.1% (38.6%, 67.1%). The mean preoperative and postoperative BCVA was (0.92±0.11) LogMAR and (0.88±0.10) LogMAR, respectively, and no significant difference was found ( t=-0.560, P=0.580). The number of anti-glaucoma medications was reduced from 2 (1, 3) before operation to 0 (0, 0) after operation.The complete success rate of surgery was 80% (40/50), and the qualified success rate was 94% (47/50). Surgical complications mainly included hyphema in 7 eyes, IOP spike in 7 eyes, and corneal edema in 3 eyes.No vision-threatening complication occurred. Conclusions:PEI+ GSL+ GT is preliminarily effective and safe for advanced PACG by reducing IOP and application of anti-glaucoma medications with few complications.

Objective:To analyze the epidemiological and clinical characteristics of children with brucellosis, and to provide a practical basis and experience for clinical diagnosis and treatment of brucellosis.Methods:Retrospective analysis was used to collect clinical data of children with brucellosis diagnosed at the Children's Hospital Affiliated to Zhengzhou University from May 2015 to October 2017, and their epidemiological characteristics, clinical manifestations, laboratory tests, and clinical diagnosis and treatment were summarized.Results:A total of 24 children were included, including 14 males and 10 females, with an average age of 6 years (1 year 2 months to 12 years old). Except February, onset throughout the year, higher incidence was from May to July (14 cases, 58.33%). The exposure history of the children was mainly exposure to cattle and sheep and consumption of beef and mutton (18 cases, 75.00%). The main clinical manifestations were fever in 24 cases (100.00%), bone and joint pain in 14 cases (58.33%), hepatomegaly in 9 cases (37.50%), splenomegaly in 7 cases (29.17%). Tube agglutination test (SAT) was positive in 21 cases (87.50%), weakly positive in 1 case (4.17%) and negative in 2 cases (8.33%). Brucella was detected in all 24 cases by microbial culture, and in 18 cases (75.00%) by blood culture. Eighteen cases (75.00%) had liver dysfunction. Thirteen cases were misdiagnosed, and the misdiagnosis rate was as high as 54.17%. Twenty-two cases had been cured after treatment, 2 cases relapsed and recovered after continued treatment. Conclusions:Children with brucellosis have diverse epidemiology and clinical features, and are easily misdiagnosed. For children with fever, bone and joint pain and exposure history, pediatricians should be alert to the possibility of brucellosis, conduct microbiological and serological tests, in order to timely, accurate and standardized diagnosis and treatment of children with brucellosis.

Zinc oxide quantum dots (ZnO QDs) were synthesized by gel-sol method and employed as the transdermal aloesin (Alo) carriers. ZnO QDs were surface-functionalized with amino using aminopropyltriethoxysilane (APTES). Alo was covalently bonded on the surface of ZnO QDs via N,N'-carbonyldiimidazole to obtain Alo NPs, which were characterized by transmission electron microscope (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR) and thermal gravimetric analyzer (TGA). TEM images showed that ZnO QDs were analogously sphere and monodisperse with a reasonably narrow size distribution, of which was around 4 nm. The size of Alo NPs increased to around 8 nm due to the surface modification. The intense bands at around 3 400 cm and 1 200 cm in the FTIR spectrum of Alo NPs from the vibration of -OH indicated the linkage of Alo on the surface of ZnO QDs. The results of TGA analysis showed that the mass ratio of ZnO QDs and Alo were 39.27% and 35.14%, respectively. The penetration of Alo NPs was much higher than raw Alo according to the passive penetration experiments with Franz-type diffusion cells instrument using full-thickness cavy skin, which manifested the improvement of the penetration for Alo delivered by ZnO QDs. The pH-controlled drug release behavior was investigated. At pH 7.4, only a small amount of Alo (1.45% ± 0.21%) had been released after 2 h. In contrast, as incubation at pH 5.0 of which pH was similar to endosomal environment, Alo was released very fast (87.63% ± 0.46% in 2 h) from Alo NPs, confirming that Alo NPs could response to the pH and realize the intracellular drug release. The inhibitory effect of Alo NPs on tyrosinase was in a dose dependent manner. When the concentration of Alo NPs was 12.5 μg/mL, the inhibition rate was up to 40.32% ± 1.57%. All the results show that the Alo NPs hold a great potential in transdermal tyrosinase inhibition.
Administration, Cutaneous ; Animals ; Chromones ; administration & dosage ; Drug Delivery Systems ; Glucosides ; administration & dosage ; Guinea Pigs ; Monophenol Monooxygenase ; metabolism ; Nanoparticles ; Quantum Dots ; Zinc Oxide

Objective To investigate the prokaryotic expression and immunoreactivity of BspE,a type Ⅳ secretion protein of Brucella,and the effect of recombinant protein BspE on cytokines.Methods According to the BspE gene of Brucella M5-90 published in GenBank,the gene fragments were synthesized by a company and then ligated into PUC57 vector for sequencing.The sequenced gene was cloned into a prokaryotic expression vector pET-28α and transformed.Induced expression was performed in E.coli DE3 competent cells.The obtained target protein was purified by a Ni-NTA affinity column,and its reactogenicity was analyzed by Western blotting.Mouse RAW264.7 cells were treated with 25 g/L BspE recombinant protein for 12,24,48 h,and the control group was treated with the same amount of BSA instead of BspE,and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-1β level.Results The recombinant expresed plasmid of pET-28α-BspE was successfully obtained.The results of Western blotting showed a single band with a relative molecular mass of about 30.1 × 103,and the recombinant protein BspE had good reactogenicity,and IL-1β levels (ng/L)were significantly elevated by the recombinant protein BspE (12 h:43.27 ± 2.13 vs 30.24 ± 1.66,24 h:57.78 ± 3.44 vs 41.22 ± 1.22,48 h:72.52 ± 3.04 vs 46.77 ± 2.75,t =8.38,7.86,10.89,P ＜ 0.05).Conclusions BspE recombinant protein has better immunoreactivity and can increase the expression level of IL-1β in mouse macrophages.This study provides a scientific basis for the role of effector proteins in the pathogenesis of Brucella.

A high performance liquid chromatography tandem mass spectrometric ( HPLC-MS/MS ) method was developed for the determination of seven perfluorinated alkyl acidin ( C4-C10 ) and perfluorooctane sulfonate in water. After the particulate was removed by leaching, surrogate standard was added, then the sample was loading to a pre-conditioned WAX cartridge for purification, and then the eluent was concentrated and analyzed by HPLC-MS/MS. Due to the situation that the fluoride polymer was unavoidable to be used in the LC system, a delay column was employed and the perfluorooctanoic acid ( PFOA ) of interference was departed from the PFOA in sample. The method detection limit ( MDL) of PFOA was 0. 8 ng/L, and the lowest quantitative concentration (LQC) was 3. 2 ng/L. For other compounds, the MDL was ranged from 0. 2 to 1. 2 ng/L, and the LQC was 0. 8-4. 8 ng/L. This method also had good reproducibility, for six duplicated samples, the relative standard deviations ( RSD ) of all target compounds were less than 16%. And the recoveries of target compounds at six spiked matrix samples ranged from 87% to 129%, and the RSD were less than 15%. Because of the connection of delay column, the background was well controlled, and a relatively lower MDL were obtained.

Objective To renovate angiography in identifying portal vein anatomy during transjugular intrahepatic portosystemic shunt (TIPS) procedures,saving the time of TIPS procedures,decreasing the risk of the complications of the post-procedure.MethodsThe difference between the Wedge hepatic venography with Carbon Dioxide in 6 cases and Inferior Mesenteric artery angiography in 7 cases during TIPS procedures were compared in the identification of portal vein anatomy.The quality of images,their effects on the procedures,the complications and the recovery post-procedure were evaluated.Results Using CO2,the portal veins were opacified in all 6 cases.TIPS procedures succeeded in all cases except 1 case because of poor coagulation function.Using Inferior Mesenteric artery angiography,the portal veins were opacified in al1 7 cases.TIPS procedure succeeded in all cases except 1 case because of chronic portal occlusion.Puncture-site hematoma occurred in 1 case after TIPS procedure.ConclusionWedge hepatic venography with Carbon Dioxide is superior,safer and more convenient than Inferior Mesenteric Artery angiography in identifying portal vein anatomy during TIPS.

Objective To investigate the protective effects of alpha-lipoie acid (ALA) against beta-cell damage in streptozotocin (STZ)-induced diabetes in rats. Methods Thirty SD rats were randomly divided into three groups: normal control (NC) group, STZ group and ALA + STZ group, with 10 rats in each group. mg/kg, intraperitoneal injection), till the end of the study (4 weeks later). Blood glucose were measured every 3 days after STZ injection. Malondialdehyde (MDA) and reduced glutathione (GSH) levels were measured in pancreatic homogenates. Pancreatic beta-cells were examined by immunohistocbemical methods, Results STZ induced a significant increase of the level of blood glucose. Body weight of rats in ALA + STZ group was (341±26)g, which significantly lower than (368±3)g in NC group, and high than (301±2)g in STZ group with stas(P < 0. 05). Meanwhile the MDA levels in STZ group and NC group were(1.22 ± 0. 14) and(0.57 ± 0.04)nmoL/mg prot, respectively, and there was significant difference between the two groups (P < 0.05) ; the GSH levels in STZ group and NC group were(16.54 ± 1.10) and(25.46 ± 0.62) mg/g prot (P < 0.05), respectively; degeneration of islet cells and decreased blood glucose were observed in STZ + ALA-pretreated rats; MDA level in pancreatic homogenates was(0.72 ± 0. 23)nmoL/mg prot, which was significantly lower than that in STZ group (P < 0.05) ; the GSH level was (35.33 ± 2.66) mg/g prot, which was significantly higher than that in STZ group (P < 0.05) ; increased staining of insulin and preservation of islet ceils functions were more obvious in the STZ + ALA-pretreated rats. Conclusions ALA exerted its protective effect through reducing the oxidative stress and preserving pancreatic beta-cell integrity.

BACKGROUND: Tissue-engineered skin plays an important role in the repair and reconstruction of large-area skin lesion. Little is known about the effects of Chinese medicine-loading tissue-engineered skin scaffold on cell adhesion and wound surface infection. OBJECTIVE: To screen the application concentration of shengji solution, which can promote tissue regeneration, using keratinocytes and fibroblasts, and to observe the effects of shengji solution-gelatin-chitosan drug-loading skin scaffold on cellular adhesion and biocompatibility. DESIGN, TIME AND SETTING: Taking keratinocytes and fibroblasts as subjects, the present randomized and controlled experiment was performed at the Laboratory of Cell Engineering, Orthopedic Institute, Tianjin Hospital between February and August 2005. MATERIALS: One healthy big-ear rabbit was included for harvesting skin seed cells. Shengji solution, Danggui (Radix Angelicee Sinensis) and Shengdi (rehmannia dride rhizome) extract (self-extracted, 1.5 g/mL), gelatin-chitosan scaffold and shengji solution-gelatin-chitosan scaffold were provided by Tianjin University, China. METHODS: Passage 3 keratinocytes and fibroblasts were harvested by dispase Ⅱ- trypsase- ethylenediamine tetraacetic acid(EDTA) digestion. The prepared cells were divided into 5 groups: control, shengji solution, Danggui, Shengdi, and epidermal growth factor (EGF). Common culture solution containing 0.1 volume fraction of fetal bovine serum, shengjisolution (5, 6, 8, and 12 g/L), Danggui extract (8 g/L), Shengdi extract (8 g/L), and EGF culture medium (10 μ g/L) were used in corresponding groups for cell culture. At 1, 3, 5, and 7 days, cellular proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. Keratinocytes and fibroblasts were cultured in different concentrations of shengji solution (50, 60, 80, and 120 g/L) and grouped according to different concentrations. At 7 and 14 days, cellular adhesion was observed by semi-quantitative method. At 4, 7, and 14 days, keratinocytes cultured by 60 and 80 g/L shengji solution were harvested for hematoxylin-eosin staining and subsequent scanning electron microscope observation. MAIN OUTCOME MEASURES: Effects of shengji solution on skin seed cell proliferation; effects of drug-loading scaffold on cellular adhesion; and cell-carrier biocompatibility. RESULTS: MIF detection results demonstrated that cells significantly proliferated after treatment of 5, 6, and 8 g/L shengji solution compared to the control group (P < 0.05). Obvious proliferation of passage 3 keratinocytes and fibroblasts was found on 60 and 80 g/L drug-loading scaffold. Keratinocytes on the drug-loading scaffold exhibited good cellular morphology and closely adhered to the scaffold. Shengji solution had no apparent toxic effects on cells. CONCLUSION: Shengji solution (60 and 80 g/L)-gelatin-chitosan scaffold can effectively promote the adhesion and proliferation of keratinocytes and fibroblasts.