Analysis of choroidal thickness and blood perfusion in idiopathic macular hole eye / 中华眼底病杂志

Objective:To observe and analyze the macular choroidal thickness and choroidal blood perfusion (CBP) in eyes with idiopathic macular hole (IMH) and their correlation.Methods:A cross-sectional observational clinical study. From March 2019 to October 2021, 60 IMH patients with 60 eyes (IMH group) and 60 healthy volunteers with 60 eyes (control group) who consecutively visited Department of Ophthalmology of The First Affiliated Hospital of Zhengzhou University were included in the study. Among the 60 eyes in the IMH group, 8, 8, 15, and 29 eyes were at stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ, respectively. There was no significant difference in age, spherical equivalent power and axial length between the two groups ( t=1.327, 0.157, 0.542; P>0.05). The average macular choriodal thickness (AMCT) and CBP in different regions of the macular region of the examined eye were measured using a swept-frequency light source optical coherence tomography scanner. According to the zoning method for the treatment of diabetic retinopathy, the choroid within 6 mm of the fovea was divided into 3 concentric circles with the fovea as the center. They are the central area with a diameter of 1 mm, the inner ring area of 1-3 mm, and the outer ring area of 3-6 mm; the inner ring area and the outer ring area were divided into 4 areas by 2 radiations respectively, including the upper part of the inner superior (IS), the lower part of the inner inferior (Ⅱ ), and the nasal side of the inner nasal (IN), inner temporal (IT), outer superior (OS), outer inferior (OI), outer nasal (ON), outer temporal (OT), a total of 9 regions. The distribution characteristics of AMCT and CBP in different regions were observed. The correlation between AMCT and CBP was analyzed by Pearson correlation; the correlation between AMCT, CBP and IMH stage was analyzed by Spearman correlation. Results:Compared with the eyes of the control group, the AMCT of the affected eyes in the IMH group was significantly thinner in all areas of the macula, and the difference was statistically significant ( t=2.378, 4.641, 2.888, 3.390, 3.575, 4.870, 4.077, 4.946, 4.578; P<0.05). Compared with the control group, the CBP in the OS and OT regions of the affected eyes in the IMH group was significantly lower, the difference was statistically significant ( t=3.424, 4.516; P<0.05). The results of Pearson correlation analysis showed that there was a significant positive correlation between AMCT and CBP in the OT region ( r=0.314, P<0.001). Spearman correlation analysis showed that there was a significant positive correlation between AMCT and IMH staging in each region ( r=0.375, 0.374, 0.289, 0.379, 0.441, 0.392, 0.303, 0.341, 0.292; P<0.05). There was no significant correlation between CBP and IMH staging in IN, OI and OT regions ( r=-0.138, -0.016, -0.221; P>0.05); CBP and IMH staging in other regions were significantly negatively correlated ( r=-0.560, -0.390,-0.819, -0.692, -0.329, -0.587; P<0.05). Conclusions:The choroidal thickness in the macular region of the eyes with IMH is significantly thinner than that of the normal subjects; there is choroidal hypoperfusion in local areas. There is a significant positive correlation between local regional AMCT and CBP; IMH stage is higher, the trend of AMCT in each region is thickening, and the CBP in most regions decrease.

Analysis of choroidal thickness and blood perfusion in idiopathic macular hole eye / 中华眼底病杂志

Objective:To observe and analyze the macular choroidal thickness and choroidal blood perfusion (CBP) in eyes with idiopathic macular hole (IMH) and their correlation.Methods:A cross-sectional observational clinical study. From March 2019 to October 2021, 60 IMH patients with 60 eyes (IMH group) and 60 healthy volunteers with 60 eyes (control group) who consecutively visited Department of Ophthalmology of The First Affiliated Hospital of Zhengzhou University were included in the study. Among the 60 eyes in the IMH group, 8, 8, 15, and 29 eyes were at stage Ⅰ, Ⅱ, Ⅲ, and Ⅳ, respectively. There was no significant difference in age, spherical equivalent power and axial length between the two groups ( t=1.327, 0.157, 0.542; P>0.05). The average macular choriodal thickness (AMCT) and CBP in different regions of the macular region of the examined eye were measured using a swept-frequency light source optical coherence tomography scanner. According to the zoning method for the treatment of diabetic retinopathy, the choroid within 6 mm of the fovea was divided into 3 concentric circles with the fovea as the center. They are the central area with a diameter of 1 mm, the inner ring area of 1-3 mm, and the outer ring area of 3-6 mm; the inner ring area and the outer ring area were divided into 4 areas by 2 radiations respectively, including the upper part of the inner superior (IS), the lower part of the inner inferior (Ⅱ ), and the nasal side of the inner nasal (IN), inner temporal (IT), outer superior (OS), outer inferior (OI), outer nasal (ON), outer temporal (OT), a total of 9 regions. The distribution characteristics of AMCT and CBP in different regions were observed. The correlation between AMCT and CBP was analyzed by Pearson correlation; the correlation between AMCT, CBP and IMH stage was analyzed by Spearman correlation. Results:Compared with the eyes of the control group, the AMCT of the affected eyes in the IMH group was significantly thinner in all areas of the macula, and the difference was statistically significant ( t=2.378, 4.641, 2.888, 3.390, 3.575, 4.870, 4.077, 4.946, 4.578; P<0.05). Compared with the control group, the CBP in the OS and OT regions of the affected eyes in the IMH group was significantly lower, the difference was statistically significant ( t=3.424, 4.516; P<0.05). The results of Pearson correlation analysis showed that there was a significant positive correlation between AMCT and CBP in the OT region ( r=0.314, P<0.001). Spearman correlation analysis showed that there was a significant positive correlation between AMCT and IMH staging in each region ( r=0.375, 0.374, 0.289, 0.379, 0.441, 0.392, 0.303, 0.341, 0.292; P<0.05). There was no significant correlation between CBP and IMH staging in IN, OI and OT regions ( r=-0.138, -0.016, -0.221; P>0.05); CBP and IMH staging in other regions were significantly negatively correlated ( r=-0.560, -0.390,-0.819, -0.692, -0.329, -0.587; P<0.05). Conclusions:The choroidal thickness in the macular region of the eyes with IMH is significantly thinner than that of the normal subjects; there is choroidal hypoperfusion in local areas. There is a significant positive correlation between local regional AMCT and CBP; IMH stage is higher, the trend of AMCT in each region is thickening, and the CBP in most regions decrease.

Application effect analysis of artificial intelligence automatic diagnosis system for diabetic retinopathy in elderly diabetic patients in community and hospital / 中华眼底病杂志

Objective:To study the efficiency and difference of the artificial intelligence (AI) system based on fundus-reading in community and hospital scenarios in screening/diagnosing diabetic retinopathy (DR) among aged population, and further evaluate its application value.Methods:A combination of retrospective and prospective study. The clinical data of 1 608 elderly patients with diabetes were continuously treated in Henan Eye Hospital & Henan Eye Institute from July 2018 to March 2021, were collected. Among them, there were 659 males and 949 females; median age was 64 years old. From December 2018 to April 2019, 496 elderly diabetes patients were prospectively recruited in the community. Among them, there were 202 males and 294 female; median age was 62 years old. An ophthalmologist or a trained endocrinologist performed a non-mydriatic fundus color photographic examination in both eyes, and a 45° frontal radiograph was taken with the central fovea as the central posterior pole. The AI system was developed based on the deep learning YOLO source code, AI system based on the deep learning algorithm was applied in final diagnosis reporting by the"AI+manual-check" method. The diagnosis of DR were classified into 0-4 stage. The 2-4 stage patients were classified into referral DR group.Results:A total of 1 989 cases (94.5%, 1 989/2 104) were read by AI, of which 437 (88.1%, 437/496) and 1 552 (96.5%, 1 552/1 608) from the community and hospital, respectively. The reading rate of AI films from community sources was lower than that from hospital sources, and the difference was statistically significant ( χ2=51.612, P<0.001). The main reasons for poor image quality in the community were small pupil (47.1%, 24/51), cataract (19.6%, 10/51), and cataract combined with small pupil (21.6%, 11/51). The total negative rate of DR was 62.4% (1 241/1 989); among them, the community and hospital sources were 84.2% and 56.3%, respectively, and the AI diagnosis negative rate of community source was higher than that of hospital, and the difference was statistically significant ( χ2=113.108, P<0.001). AI diagnosis required referral to DR 20.2% (401/1 989). Among them, community and hospital sources were 6.4% and 24.0%, respectively. The rate of referral for DR for AI diagnosis from community sources was lower than that of hospitals, and the difference was statistically significant ( χ2=65.655, P<0.001). There was a statistically significant difference in the composition ratio of patients with different stages of DR diagnosed by AI from different sources ( χ2=13.435, P=0.001). Among them, community-derived patients were mainly DR without referral (52.2%, 36/69); hospital-derived patients were mainly DR requiring referral (54.9%, 373/679), and the detection rate of treated DR was higher (14.3%). The first rank of the order of the fundus lesions number automatically identified by AI was drusen (68.4%) and intraretinal hemorrhage (48.5%) in the communities and hospitals respectively. Conclusions:It is more suitable for early and negative DR screening for its high non-referral DR detection rate in the community. Whilst referral DR were mainly found in hospital scenario.

Analysis of BEST1 gene mutation and clinical phenotype in two families with Best vitelliform macular dystrophy and autosomal recessive bestrophinopathy / 中华眼底病杂志

Objective:To report the BEST1 gene mutations and clinical phenotypes in two pedigrees with Best vitelliform macular dystrophy (BVMD) and autosomal recessive bestrophinopathy (ARB). Methods:A retrospective clinical study. From November 2019 to March 2021, in the Department of Ophthalmology of The First Affiliated Hospital of Zhengzhou University, the BVMD family (4 patients and 6 family members) and the ARB family (2 patients, 2 family members), a total of 6 patients and 8 normal family members were included in the study. Detailed medical history was obtained; best corrected visual acuity, fundus color photography, electrophysiology, optical coherence tomography and fundus autofluorescence examination were performed. The clinical characteristics for all patients in the two families were analyzed. Three milliliter peripheral venous blood of all participants in the family was collected, and the whole genomic DNA was extracted with gene sequencing using next-generation sequencing technology based on targeted capture. Compared with the database to identify the pathogenicity mutation sites, suspected pathogenic mutation sites were selected, then mutations in other members in the family was assayed by Sanger sequencing.Results:In family 1, the proband was demonstrated as typical BVMD, other patients were multifocal vitelliform macular dystrophy. The DNA sequencing result showed that all the 4 patients carried heterozygous missense mutations in exon 3 of BEST1 gene: c.240C>G (p.F80L) (M1) and 2 members carried this mutation, but without clinical phenotype. M1 was a likely-pathogenic mutation reported for the first time. In family 2, the proband and the other patient were diagnosed as ARB. The DNA result showed that the 2 patients carried heterozygous missense mutations in exon 5 and exon 2 of BEST1 gene: c.584C>T (p.A195V) (M2)、c.139C>A (p.R47S) (M3), and a heterozygous frameshift mutation in exon 3 of BEST1 gene: c.235dupT (p.S79Ffs*153) (M4). M2 was a pathogenic mutation reported previously. M3 variant was of undetermined significance. M4 was a first reported pathogenic mutation. Conclusions:The BEST1 gene mutation is the main cause of BVMD and ARB. Different mutation sites have different clinical phenotypes. BVMD and ARB have genetic and clinical heterogeneity.

Analysis of USH2A gene mutation and clinical phenotype in families with Usher syndrome type 2 and retinitis pigmentosa / 中华眼底病杂志

Objective:To observe the gene mutations and clinical phenotypes in patients with Usher syndrome type 2 (USH2) and retinitis pigmentosa (RP).Methods:From August 2018 to January 2019, 4 patients and 11 normal family members from 3 families of USH2 and RP who visited Henan Eye Hospital were enrolled in the study. Detailed medical history was obtained and visual acuity, fundus color photography, OCT, visual field, full field ERG examination were performed. Among the three families, pedigree 1 was diagnosed with USH2, pedigree 2 and pedigree 3 were diagnosed with RP. The peripheral venous blood of patients and their family members were collected, and the whole genomic DNA was extracted. Targeted capture next generation sequencing analysis was performed on these members, and Sanger sequencing and family cosegregation were verified.Results:In the family F1, the proband had symptoms of RP and sensorineural deafness. Sequencing revealed two heterozygous frameshift variants: c.13877-13880 del AGAC (p. Q4626P) in exon 64 and c.798 del T (p. F266L) in exon 5 of USH2A. Both patients of family 2 and 3 showed RP signs without deafness. Two heterozygous variants c.15178T> C (p. S5060 P) in exon 70 and c.6986C> A (p. P2329H) in exon 37, and a pathogenic heterozygous variant c.5836C> T (p. R1946X) in exon 29 of USH2A were identified in family F2. A heterozygous missense variant c.14951C> T (p. P4984L) in exon 68 and a variant c.11156G> A (p. R3719H) in exon 57 of USH2A were found in family F3. The results of conservation analysis showed that the corresponding amino acid sites of USH2A p.Q4626P, p.F266L, p.S5060P, p.P2329H and p.P4984L were highly conserved in many species. Among these 7 pathogenic variants detected, M1-M4 and M6 were novel.Conclusions:Mutation USH2A gene are the main cause of USH2 and non-syndromic RP. Different variants affect protein translation and synthesis, consequently causing different clinical phenotypes.

Expression of metastasis associated genes nm23, p53 and S100A4 in conjunctival melanoma and their relationship with invasion and metastasis / 中华实验眼科杂志

Objective To study the significance of metastasis associated genes nm23,p53 and S100A4 in the development of conjunctival melanoma metastasis.Methods Conjunctival melanoma tissue specimens were collected from 42 cases of conjunctival melanoma patients in Henan Eye Hospital from July 2015 to November 2016,meanwhile 30 cases of conjunctival nevus tissue samples served as control group under the informed consent.The expressions of nm23,p53 and S100A4 were detected in conjunctival melanoma group and control group by Western blot and immunohistochemical method.The relationship between the clinical and pathological features of nm23,p53 and S100A4 with conjunctival melanoma patients with lymph nodes metastasis were analyzed.Results Western blot assay showed that the expression level of nm23 in conjunctival melanoma group was lower than that in the control group,with a significant difference between them (P＜0.05);the expression level of p53 and S100A4 in conjunctival melanoma group was significantly higher than those in the control group (both at P＜0.05).Immunohistochemistry showed that nm23 and S100A4 appeared claybank in cytoplasm,while p53 appeared red in cell nucleus.The positive rate of nm23 protein expression in conjunctival melanoma group was significantly lower than that in the control group (14.3％ vs.53.3％,P＜0.05).The positive rate of p53 expression in conjunctival melanoma group was significantly higher than that in the control group (54.8％ vs.6.7％,P＜0.05).The positive rate of S100A4 expression in the conjunctival melanoma group was significantly higher than that in the control group (59.5％ vs.6.7％,P＜0.05).There were no difference in nm23,p53 and S100A4 protein expression positive rate between various gender groups,various age groups and various ulcer groups (all at P＞0.05).There were siginificant differences in nm23,p53 and S100A4 protein expression positive rate between various sclera invasion groups and various distant metastasis groups (all at P＜0.05).Conclusions Expression of nm23,p53 and S100A4 may play an important role in the invasion and metastasis of conjunctival melanoma,which may be the main reference index for the diagnosis and treatment.

Removal of protein extract from moringa oleifera seed presscake on turbidity water / 中国生化药物杂志

Objective To study the removal effect of Moringa seedprotein on turbidity water. Methods The protein of Moringaoleifera seed was extracted by salting out and salting out methodand the protein concentration of Moringa oleifera seed wasdetermined by Coomassie blue staining;The removal effect of Moringa seed protein on turbidity water was determined by coagulation test.The contents of COD, ammonia nitrogen and nitrate nitrogen in the water were determined to determine the effect of Moringa seed protein on water quality. Results The experimental results showed that Moringa oleifera seed protein has good removal effect on high and medium turbidity water, and its removal effect is in a dose - dependent manner.The removal rate of 7 mg/L of Moringa seed protein to high and medium turbidity water reached 92.25 ％ and 64.71 ％ respectively. But the removal efficiency of low turbidity water was less than 7 mg/L, the removal rate of low turbidity water was only 31.91％.The results of determination of COD, ammonia nitrogen and nitrate nitrogen showed that the Moringa seed protein did not increase the content of organic matter in the water while removing turbidity effectively. Conclusion Moringa oleifera seed protein has a certain removal effect on turbidity water, among which the removal effect of high and medium turbidity water is strong, and the removal effect of low turbidity water is poor.Moringa oleifera seed protein had little effect on water quality.

Effects of adiponectin on proliferation, migration and tube formation of RF/6A cells / 眼科新进展

Objective To investigate the effects of adiponectin (APN) on proliferation,migration and tube formation of RF/6A cells,and explore the effects of APN on choroidal and retinal angiogenesis.Methods Well cultured RF/6A cells were randomly divided into the control group and three groups with different concentrations of recombinant adiponectin (5 pg · mL-1,50 pg · mL-1,500 pg · mL-1) for 1 hour.After 24 hours,cell proliferation,migration and tube formation were detected by MTT assay,wound scratch assay and seeding cells in matrigel,respectively.Results The cell proliferation in all APN groups was weaker than that of control group (P ＜ 0.05),the cell migration area (pixels) of all APN groups was significantly smaller than that of the control group (P ＜ 0.05),and the number of tube formation of all APN groups was significantly less than the control group (P ＜ 0.05).Cell proliferation,migration and tube formation decreased along with the increase of APN concentration.Conclusion APN can obviously inhibit the angiogenesis process of RF/6A cells.This inhibitive effect indicates the protective role of APN in choroidal and retinal angiogenesis.

Inhibition of gallnut extract on MRSA β-lactamase / 中国生化药物杂志

Objective Study on the inhibitory effect of gallnut extract extract on MRSA β-lactamase. Methods Determination of inhibitory effect of gallnut extract on MRSA3002 by TTC method. β-lactamase was repeated by freezing and thawing method . Synergistic effect of gallnut extract and gentamicin was detected by TTC. Results The MIC and MBC of MRSA3002 by gallnut extract were 8mg/mL and 32mg/mL.Gallnut extract can reduce strains of β-lactamase activity,the MRSA300224h 1/2MIC after the effect of gallnut extract, beta lactam enzyme activity inhibition compared with the control group there were significant differences (P<0.01),compared with the positive control group, the difference was not significant. Synergistic effect of gallnut extract and gentamicin can significantly reduce the MIC of MRSA3002. Conclusion Gallnut extract can reduce β-lactamase activity recovery sensitivity of drug-resistant bacteria.

The antimicrobial and antioxidant activity of the powder of Ganoderma lucidum / 中国生化药物杂志

Objective To investigate the antimicrobial and antioxidant activity of the water extract and ethanol extract of the powder of G.lucidum.Analysis and comparison of two kinds of antibacterial and antioxidative effects of the extracts.Methods G.lucidum powder was prepared by using a pulverizer and extracted with water and ethanol respectively.The inhibitory effects of water extract and alcohol extract of G.lucidum on the tested strains were determined by TTC method.The antioxidant activities of aqueous extract and ethanol extract of G.lucidum were determined by DPPH and FRAP.Results The minimum inhibitory concentration ( MIC) of the aqueous extracts and ethanol extracts of G.lucidum were 32 mg/mL and 16 mg/mL, respectively.The MIC for S.Aureus respectively was 64 mg/mL and 32 mg/mL, the MIC for M.luteus respectively was 32 mg/mL and 8 mg/mL, MIC for B.terom respectively was 64 mg/mL and 8 mg/mL, The MIC of B.subtilis respectively was 32 mg/mL and 8 mg/mL.The antioxidant activity of G.lucidum powder extract and ethanol extract showed good antioxidant activity, the scavenging radical capacities on DPPH of the water extract and ethanol extract were 56.22%and 20.67% at a concentration of 2 mg/mL ( P<0.5 ) , the FRAP value respectively were 3.52 and 3.77 mmol/mL. Conclusion The powder of G.lucidum has the effective antimicrobial activity and antioxidant activity.The two kinds of extracts extract of G.lucidum had good antibacterial activity and antioxidant activity.The ethanol extract of G.lucidum powder antimicrobial effect and total antioxidative ability in water extracts, aqueous extract of Ganoderma lucidum granules on DPPH free radical scavenging rate better than that of ethanol extract.

Effect of ethanolic extract of Omithogalum caudatum Ait on apoptosis of Candida albicans / 中国生化药物杂志

Objective To investigate the effect of Omithogalum caudatum Ait(OCA)on apoptosis of Candida albicans,illustrated the antifungal mechanism of OCA.Methods Annexin Ⅴ-FITC/PI double stainingwas used to detect the effect of OCA on the apoptosis of C.albicans;JC-1 and DCFH-DA staining were used to detectthe effect of OCA on mitochondrial membrane potential(MTP)and reactive oxygen species(ROS)of C.albicans.Results OCA had a good antifungal activity,the minimum inhibitory concentration(MIC)and the minimum fungicidal concentration(MFC)were 8mg/mL and 32mg/mL respectively.OCA could induceapoptosis of C.albicans,promote the reduction of MTP and increase of ROS.Conclusion OCA induced cell apoptosismainly through disrupting mitochondrial function.

Inhibition mechanism of gallnut on biofilm formation by methicillin-resistant Staphylococcus aureus / 中国生化药物杂志

Objective To investigate the inhibition mechanism of gallnut on biofilm formation by MRSA 41577.Methods TTC assay was used to detect inhibitory effects of biofilms formation and mature biofilms.The of PIA on biofilm formation was studied using Congo red agar method.Micro-Ultraviolet Spectrophotometer was used to detect inhibitory effects of the release of eDNA.The influence for Baicalein on icaA and cidA gene expression were detected by RT-PCR method.Results The inhibitory concentration (MIC) and minimum bactericidal concentration (MIC) of MRSA 41577 BF were 0.5 mg/mL and 1 mg/mL, respectively.The inhibitory effect of galla on MRSA 41577BF formation and mature BF was significantly inhibited.Inhibition of MRSA 41577,the MIC and MBC of mature BF were 4 mg/mL and 16 mg/mL.Congo red test results show that Galla can inhibit the synthesis of MRSA 41577 PIA, and the concentration was dose-dependent.The results showed that gallnut could inhibit the release of MRSA 41577 eDNA, and the release amount of eDNA was 3.61μg/OD595 and 11.91μg/OD595 , respectively, when the concentration of gall was 1/2MIC.The release of eDNA was reduced by 69.7% (P<0.01).The expression of icaA and cidA genes in the control group was 9.7% and 6.67%, respectively.The expression of icaA and cidA in the control group was significantly lower than that in the control group ( icaA and cidA, and cidA gene expression were 100%, the expression of icaA and cidA genes were reduced by 90.3%and 93.3%, respectively (P<0.01).Conclusion The inhibitory effect of gallnut on the biofilm of MRSA 41577 is mainly through inhibiting the expression of icaA and cidA genes, and then affecting the synthesis of PIA and the secretion of eDNA .

Compare anti-tumor activity and antioxidant activity of F.velutipes and A.auricula / 中国生化药物杂志

Objective To compare the anti-tumor activity and antioxidant activity of ethanol extracts of F.veluties and A.auricula.Methods Three human carcinoma cell lines, including MCF-7, HeLa and A375 were assessed by MTT assay to measure cell viability.The antioxidant activity was detected with a DPPH and RFAP assays.Results Two domestic fungus have different anti-tumor activity on the inhibition of MCF-7, HeLa and A375 cells at 25-400μg/mL concentration rage.F.velutipes was better than the A.auricula.The difference was statistically significant ( P<0.05 ).Compared with the control group, the inhibition of F.velutipes were 48.20%, 52.61%and 50.58%at the concentration of 400μg/mL, respectively (P<0.01).At the same concentration, the inhibition of A.auricula were 37.62%、50.21%and 41.59%, respectively (P<0.01).The ethanol extracts of two domestic fungus have significant antioxidant activity.F.velutipes was better than the A.auricula.The difference was statistically significant ( P<0.01 ).Compared with the control group, the scavenging radical capacities on DPPH of ethanol extracts of F.velutipes and A.auricula were 60.30%and 40.43%at the concentration of 1.6 mg/mL, respectively (P<0.01).At the same concentration FRAP value were 9.5 and 7.0 mmol/mL(P<0.01).Conclusion The F.velutipes and A.auricula both have strong anti-tumor and antioxidant activities, F.velutipes is better than A.auricula.

Inhibitory effects of andrographis paniculata and silybum marianum on the efflux pump of MRSA 41577 / 中国生化药物杂志

Objective To study the inhibitory effects of andrographis paniculata and silybum marianum on the efflux system of MRSA 41577. Methods Inhibitory effects of andrographis paniculata and silybum marianum on efflux system of MRSA 41577 was evaluated using fluorescence spectrophotometry.PCR was applied to detect the norA efflux gene.By RT-PCR method for detection of andrographis paniculata and silybum marianum influence of the expression of norA efflux gene.Results Andrographis paniculata and silybum marianum significantly increased the accumulation of ciprofloxacin in MRSA 41577 in a time-dependent manner.At 12 minute, andrographis paniculata and silybum marianum respectively increased ciprofloxacin in MRSA41577 by 49% and 76%( P <0.05 ) , which is superior to that of reserpine. Further mechanism studies indicated that andrographis paniculata and silybum marianumcould reduce the expression of norA in MRSA 41577.After incubated with andrographis paniculata and silybum marianum for 16 h, the relative expression of norA of MRSA41577 was respectively reduced by 35% and 42% ( P <0.05 ). Conclusion Andrographis paniculata and silybum marianumcould inhibit MRSA efflux system through reducing pathogen ’s expression of norA and NorA protein.

Inhibitory effects of Sanguisorba officinalis L.on MRSA biofilms formation / 中国生化药物杂志

Objective To investigate inhibitory effects of Sanguisorba officinalis L.on biofilms of MRSA41577.Methods Congo red agar method and crystal violet semi-quantitative method were used to detect the biofilms-forming ability of tested strains; TTC assay was used to detect inhibitory effects of Sanguisorba officinalis L.on biofilms formation and mature biofilms of MRSA41577,as well as effects of Sanguisorba officinalis L.in combination with vancomycin on mature biofilms of MRSA41577.Results Sanguisorba officinalis L.showed significant inhibitory both on biofilms formation and mature biofilms, minimum inhibitory concentration(MIC) and minimal bactericidal concentration(MBC) of biofilms formation were 1 mg/mL and 8 mg/mL,MIC of mature biofilms was 4 mg/mL.The sensitivity of mature biofilms to vancomycin was greatly increased when Sanguisorba officinalis L.was combined with vancomycin with subinhibitory concentrations.Sanguisorba officinalis L.at 1/4 MIC can inhibit mature biofilms when combined with vancomycin at 4 μg/mL, while vancomycin didn't show inhibitory effects on mature biofilms when concentrations were below 64 μg/mL. Conclusion Sanguisorba officinalis L.has significant inhibitory on biofilms formation, the mechanism may be related to Sanguisorba officinalis L.destroyed biofilms and make vancomycin penetrate into the biofilms to finish the bactericidal activity.

Effect of emodin on mice immune cells and its hemolysis in vitro / 中国生化药物杂志

Objective To investigate effect of emodin on mice immune function and its hemolysis toxicity.Methods The mouse specific immune cells of T, B lymphocytes and nonspecific immune cell of macrophages and NK cells were prepared and incubated in vitro.The different immune cells were treated by emodin with different concentrations of 5,10,15 and 20μM, and DMSO as control group.The effect of emodin on immune cells function was detected by neutral red assay and MTT assay.The hemolysis test in vitro was conducted by emodin with different concentrations of 20, 40, 60 and 80μM, physiological saline as blank control group and sterile distilled water as positive control group, then the hemolysis toxicity of emodin was observed. Results There were no significant difference of T and B lymphocyte proliferation among control group, 5, 10, 15 and 20 μM group(F=0.009,P=1.000;F=0.003,P=1.000), the phagocytic ability of macrophages enhanced in each dose group and was concentration dependent(F=665.525,P=0.000), the proliferation rate of macrophages enhanced and was concentration dependent(F=134.812,P=0.000), the activity of NK cells enhanced and was concentration dependent(F=200.190,P=0.000).Hemolysis test results showed the hemolysis rate was less than 5% in the range of 20 to 80μM emodin.Conclusion Emodin could significantly promote the nonspecific immune cells activity.Within the concentration of experiment, emodin has no hemolysis toxicity.

Antibacterial activity and mechanism of baicalein / 药学学报

Baicalein (BAI) is an effective bactericide. The antibacterial activity and mechanism experiments were carried out by determining conductivity and content of macromolecules of membrane penetrability, the oxidative respiratory metabolism and protein synthesis changes and the inhibition of DNA topoisomerase activities. Electrical conductivity and the number of large molecules of BAI increased 2.48% and 1.8%, respectively, than that of the control. However, the membrane integrity did not destroyed by BAI directly. With BAI treatment, inhibition rates of activities for SDH and MDH were 56.2% and 57.4%, respectively, demonstrating that BAI could inhibit cell respiratory. After treated with BAI for 20 h, the total soluble content of proteins decreased by 42.83%. Moreover, the activities of DNA topoisomerase I and II were inhibited completely by 0.2 mmol x L(-1) BAI. These results indicated that BAI had obvious antibacterial activity on Staphylococcus aureus. The mechanism is that it could affect bacterial membrane penetrability, inhibit protein synthesis and influence SDH, MDH and DNA topoisomerase I and II activities to exert its antibacterial functions.