Background The early disorder of diabetic retinopathy (DR) is the damage of retinal neural cells induced by high glucose and lack of oxygen.Previous studies show that bushenhuoxue serum can enhance the activity of glutamine synthetase (GS) in Müller cells under hypoxia,and glutamate-mediated retinal excitotoxicity also can be reduced by bushenhuoxue serum intervention.However,whether the concentration of glycine can be increased by bushenhuoxue serum is not clear.Objective This study was to investigate the protective effects of bushenhuoxue serum on retinal ganglion cells (RGCs) under hypoxia.Methods The Sprague Dawley (SD) rat serum containing bushenhuoxue was prepared.The RGCs of newborn SD rats were purified and identified by a twostep immunopanning procedure.After 72 hours,all RGCs were cultured in 96-well plates and divided into four groups:normal control group (cultured in adult SD rats normal serum),bushenhuoxue group (cultured in bushenhuoxue serum),hypoxia group (cultured in 1 mmol/L sodium dithionite); hypoxia + bushenhuoxue intervention group (cultured in bushenhuoxue serum+sodium dithionite).Glutamate and glycine contents in the extracellular fluid were detected by L-8800 automatic amino acid analyzer,and the content of lactate dehydrogenase (LDH) was assayed using LDH kits in 24,48 and 72 hours after culture.Results Cultured cells showed the green fluorescence under the immnofluorescence microscope.The contents of glutamate,glycine and LDH in the extracellular fluid were (0.0805±0.0010)mg/L,(0.0554±0.001 5)mg/L and (1 626.03 ±122.10)μmol/(min · L) in the normal control group in 24 hours after culture,and those in the hypoxia group were (0.022 5±0.001 1) mg/L,(0.014 6±0.001 1)mg/L and (1 458.68±94.23)μμmol/(min · L),showing significant reducing in the hypoxia group (q =-3.53,P =0.00 ; q =-2.45,P =0.00 ; q =-2.98,P =0.02).Compared with the normal control group,LDH and glycine contents in the extracellular fluid were significant raised in the hypoxia group 48 hours after culture (q =2.55,P =0.01 ;q =4.48,P =0.00).Seventy two hours after culture,the glutamate and glycine contents in the hypoxia group were higher than those of the normal control group (q =2.45,P =0.00 ;q =3.72,P =0.00).In 48 and 72 hours of culture,the contents of glycine were (0.017 4±0.001 5) and (0.019 2±0.001 2) mg/L in the hypoxia+bushenhuoxue intervention group,which were significantly higher than (0.016 0±0.001 2) and (0.018 0±0.000 8) mg/L in the hypoxiagroup (q=2.28,P=0.04;q=2.33,P=0.03),but the LDH level were (1 632.94±264.31) and (1 875.00±137.45)μmol/(min · L) in the hypoxia+ bushenhuoxue intervention group,which were lower than (1 688.49 ± 112.86) and (2 267.86 ± 175.21) μmol/(min · L) of the hypoxia group (q =-2.95,P =0.02 ; q =-2.35,P=0.00).No significant differences were seen in the glutamate content 24,48 and 72 hours after culture (P=0.55,0.28,0.46).A positive correlation was seen between the glutamate content and glycine content in the extracellular fluid (Kendall coefficient =0.519,Spearman coefficient =0.696,both at P =0.000).Conclusions The release levels of glutamate and glycine increase in the hypoxia RGCs,which probably is a compensatory response of RGCs.Bushenhuoxue serum can protect RGCs against injury by increasing the release of glycine and decreasing the LDH leakage from RGCs.

Objective To study the middle ear mucosa of guinea pigs with tympanosclerosis by electron microscope and try to obtain some insights into the feature and pathogenesis of tympanosclerosis.Methods Eight healthy variegated guinea pigs were devided into 2 groups.Six(8 ears) guinea pigs were subjected to inoculation of 1×10~8/L of staphylococcus aureus solution 100 μl into the middle ear cavities under the microscope.All the guinea pigs were observed for more than 6 months with no farther treatment.For electron-microscopic studies,the mucosa tissues were taken from the tympanic mucosa in 6 guinea pigs (8 ears) with tympanosclerosis from various sites,while the middle ear mucosa of two healthy guinea pigs (4 ears) were taken as a control.Results Uhrastructural examination of the normal middle ear mucosa revealed a few collagen fibers,normal morphous of fibrocyte,rough endoplasmic reticulum and mitochondria,and there was no lysosome.However,the tympanosclerosis specimens showed that irregular deformation,elongation,and degeneration of fibrocytes and oval nucleus were darkly stained,lots of mitochondria and lysosomes gathered into the cytoplasm around the nuclear and cystic expansion of the endoplasmic reticulum.In the submucosa extracellular matrix,there were a large number of collagen fibers containing lots of amorphous high-density electron-rich body.Conclusion Electron-microscopic studies of the middle ear mucosa of guinea pigs with tympanosclerosis revealed evident proliferation of collagen fibers,and calcifications were seen in the structures such as extracellular matrix vesicle,lysosomes,myelin structures within lipid granules,which mainly in extracellular matrix vesicles.

Objective To study the synergistic antitumor effects of quercetin and cisplatin in human prostate cancer PC3 cells and LNCaP cells.Methods Twelve h after PC3 cells or LNCaP cells were seeded,different dose of quercetin (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L,160 μmol/L) or cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L),or quercetin (20 μmol/L) + cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L) were added for48 h and then the antiproliferative effects were detected with MTT assay.After incubated with quercetin (20 μmol/L) or cisplatin (0.05 μ mol/L),or quercetin (20 μ mol/L) + cisplatin (0.05 μmol/L) for48 h,cell cycle distribution and apoptosis of PC3 cells or LNCaP cells were detected by flow cytometer,PI and Annexin V staining.Protein expression was detected by Western blotting.Results After treatment with quercetin or cisplatin alone,the IC50 were (0.99 ± 0.13) μmol/L,(0.75 ± 0.09) μmol/L and (91.60 ± 6.10) μ mol/L,(72.90±4.70) μ mol/L for LNCaP cells or PC3 cells,respectively;The IC50 were (0.11±0.06)μ mol/L,(0.07±0.02) μmol/L for quercetin + cisplatin treatment (Compared with quercetin,P＜0.01 ;Compared with cisplatin,P＜0.05.After treatment with cisplatin or quercetin + cisplatin for 48 h,the S phase percent of LNCaP cells or PC3 cells were (22.4±2.7)％,(31.2±2.4)％ and (20.1±1.6)％,(31.0±2.5)％,respectively,(Compared with control,P＜0.05,however,treatment with quercetin alone has no significant difference (Compared with control,P＞0.05).After treatment with cisplatin or quercetin + cisplatin for 48 h,the apoptotic percent of LNCaP cells or PC3 cells were (14.8 ± 1.9) ％,(39.6 ± 3.1) ％ and (11.5± 1.2) ％,(34.1 ±3.3) ％,respectively,(compared with control,P ＜ 0.05,however,treatment with quercetin alone had no significant difference (compared with control,P＞0.05).After treatment with quercetin alone for 48 h,the activation of caspase-3,caspase-8 and caspase-9 were slightly increased,the expressions of Bax and p21 were up-regulated,the expressions of Bcl-2 and CDK2 were down-regulated.Furthermore,these effect of cisplatin and quercetin + cisplatin were significantly enhanced (compared with quercetin,P＜0.05;compared with quercetin,P＜0.01,respectively.Conclusions The combination modality with quercetin and cisplatin has a better treatment effect in vitro not only in androgen-dependent LNCaP cells but also in androgen-independent PC3 cells.

To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Decidua/metabolism ; Embryo Implantation/*physiology ; Endometrium/metabolism ; I-kappa B Proteins/*biosynthesis ; NF-kappa B/*biosynthesis ; Time Factors ; Uterus/*metabolism ; Uterus/physiology

BACKGROUND:Airborne pollen is the most important factor to induce the seasonal allergic diseases.The production and dispersal of pollen are closely correlated with the meteorological factors.OBJECTIVE:To observe the correlation between the seven meteorological factors (air pressure,air temperature,humidity,precipitation,wind speed,evaporation capacity and sunshine) and the airborne pollen in Nanchang so as to provide theoretic evidences for preventing and treating anaphylactic disease in that region.DESIGN:Observational experiment. SETTING:Department of Allergic Reaction,Jiangxi Medical College.MATERIALS:An investigation on airbome pollen was performed for a year by Durham gravity method.Data of airborne pollen and the seven meteorological factors in urban area of Nanchang city were collected.METHODS:The top of the 4th experimental building in the southern yard of Jiangxi Medical College located in the center of Nanchang city was taken as exposed point.The pollen was surveyed by Durham gravity method from April 1st,2000 to March 31 st,2001.Meteorological data of the seven factors were collected every day and supplied by the Weather Bureau of Jiangxi Province.The correlation between meteorological factors and airborne pollen was analyzed by the multivariate Jinear stepwise regression analysis with SAS 6.12.MAIN OUTCOME MEASURES:The type,amount,seasonal distribution of airborne pollen and the correlation between the seven meteorological factors and the airborne pollen in urban area of Nanchang city. RESULTS:There were airborne pollens in Nanchang city all the year round,which included about 47 types.The main pollen season of Cupressaceae was in March,Pinaceae in March and April, and Cunninghamia R.Br in March,Chenopodium-Amaranthus and Ambrosia from August to November, Artemisia from September to November and Humulus in September.Gramineae had been scattering all seasons but mostly from June to October.In the correlation analysis between dispersal of airbome pollen and seven meteorological factors, multivariate linear stepwise regression analysis was conducted in some major polien counting in steady dispersal period and seven meteorological factors. The positive correlation was found in Pinaceae with air pressure,air temperature,wind speed and sunlight,and the negative correlation was found with precipitation and evaporation capacity.The positive correlation was found in Cupressaceae with air pressure,air temperature and sunlight,and negative correlation was found with humidity,precipitation,wind speed and evaporation capacity.The positive correlation was found in Ambrosia with air pressure,air temperature and wind speed,while negative correlation was found with humidity,precipitation,evaporation capacity and sunshine.The positive correlation was found in Artemisia with air pressure,air temperature and wind speed,while negative correlation was found with humidity,precipitation, evaporation capacity and sunshine.The positive correlation was found in Gramineae with air pressure and air temperature,while negative correlation was found with humidity,precipitation and evaporation capacity.However,if using count of total pollen in a year or half a year to analyze the relationship,the correlated factors were much fewer than that mentioned above.CONCLUSION:The dispersal of airborne pollen is related with the seven meteorological factors.It is better to use the grain amount of single pollen in its main pollen season to analyze the relationship with meteorological factors.

Objective To analyze the effect of hand hygiene compliance on the incidence of ventilator-associated pneumo-nia(VAP )in children,and explore the role of hand hygiene in the prevention and control of VAP .Methods Hand hygiene compliance among HCWs in a pediatric intensive care unit (PICU)was intervened,hand hygiene compliance rate of HCWs and incidence of VAP in children with mechanical ventilation from March 1,to October 31,2012 (before intervention)and from January 1,to August 31,2013 (after intervention)were compared.Results Hand hygiene compliance rate after in-tervention was higher than before intervention(37.31%[97/260]vs 24.17% [58/240])(χ2 =10.08,P <0.01);the con-sumption of hand washing liquid and alcohol-based handrub enhanced from 3.48mL /bed day before intervention to 4.49 mL/bed day after intervention,which increasing by 29.02%.The incidence of VAP after intervention was lower than be-fore intervention (2.21‰[5/2 261]vs 6.50‰[13/2 001])(χ2 =4.67,P <0.05).Conclusion Implementation of com-prehensive measures and improvement of HCWs’hand hygiene compliance can reduce the incidence of VAP in chil-dren.

Objective To explore hereditary phenomenon in the process of pregnancy and childbirth .Methods Pregnancy and delivery data of pregnant women and their mothers conformed to the standard was collected from January 2013 to December 2013 for correlation analysis .Results An association existed between pregnant women and their mothers in gestational age ,weight gain during pregnancy ,child′s birth weight ,duration of labor ,delayed pregnancy experience .Conclusion Hereditary exists in the process of pregnancy and childbirth .

OBJECTIVETo investigate the clinical value of Paroxetine combined with mid-frequency electrical pulse acupoint stimulation (EPAS) in the treatment of premature ejaculation (PE).

METHODSTotally 69 PE patients were equally assigned to receive oral Paroxetine 20 mg/d, mid-frequency EPAS, or oral Paroxetine 10 mg/d combined with mid-frequency EPAS (P + EPAS) , all for 8 weeks. We obtained the intravaginal ejaculation latency time (IELT) and Chinese Index of Premature Ejaculation (CIPE-5) scores of the patients before and after treatment, and compared adverse reactions among the three groups of patients.

RESULTSOne patient of the Paroxetine group gave up treatment because of abdominal pain and nausea. Compared with the baseline, the patients in the Paroxetine, EPAS, and P + EPAS groups all showed markedly increased IELT ([0.92 ± 0.11] vs [4.07 ± 0.11] min, P < 0.01; [0.92 ± 0.12] VS [2.78 ± 0.17] min P < 0.05; [0.91 ± 0.09] vs [5.31 ± 0.13], P < 0.01) and decreased CIPE-5 scores (12.5 ± 3.0 vs 22.0 ± 2.1, P < 0.01; 12.8 ± 2.9 vs 19.5 ± 1.9, P > 0.05; 13.1 ± 2.8 vs 25.2 ± 2.1, P 0.01), with statistically significant differences between the P + EPAS group and the other two (P < 0.05). The total effectiveness rate was 95.7% in the P + EPAS group, remarkably higher than in the Paroxetine (72.7%, P < 0.05) and the EPAS group (47.8, P < 0.01).

CONCLUSIONOral Paroxetine combined with mid-frequency EPAS has a higher safety and efficacy than either Paroxetine or EPAS alone in the treatment of PE.

Acupuncture Points ; Aged ; Combined Modality Therapy ; methods ; Ejaculation ; Electroacupuncture ; methods ; Humans ; Male ; Paroxetine ; therapeutic use ; Premature Ejaculation ; therapy ; Serotonin Uptake Inhibitors ; therapeutic use ; Treatment Outcome

OBJECTIVE To investigate the prevalence of the antimicrobial and disinfectant-resistant genes from the clinical isolates of meticillin-resistant Staphylococcus aureus(MRSA) and meticillin-resistant Staphylococcus haemolyticus(MRSH).METHODS The identification and antimicrobial susceptibility test of MRSA and MRSH isolates were determined by MicroScan auto SCAN4.PCR was used for detecting antimicrobial and disinfectant-resistant genes.The minimal inhibitory concentration(MIC)and minimal bactericidal concentration(MBC)of benzalkonium bromide to staphylococci that carried qacA/B gene were determined by broth dilution method.RESULTS The positive rates of qacA/B,aac(6′)/aph(2″),aph(3′)-Ⅲ,ant(4′,4″),TEM,erm,and tetM from 24 MRSH strains were 37.5%,87.5%,33.3%,29.2%,95.8%,and 94.4%;and 91.7%.respectively,and 30.6%,91.7%,72.2%,8.3%,100%,94.4% and 91.7%,respectively.The MIC of benzalkonium bromide for MRSH and MRSA were both 32-128 mg/L,MBC for MRSH was 256-512 mg/L and for MRSA was 512-1024 mg/L.The MIC and MBC of benzalkonium bromide for standard strain ATCC25923 were 16 mg/L and 32mg/L,respectively.CONCLUSIONS Antimicrobial and disinfectant-resistant genes are commonly prevalent in MRSA and MRSH isolates.

Purpose To investigate the clinicopathological characteristics and treatment of combined hepatocellular carcinoma-cholangio carcinoma (cHCC-CC).Methods 24 cases of cHCC-CC were collected.The clinical pathological characteristics,imaging,immunophenotyping and clinical features were retrospectively analyzed and reviewed the literature.Results There were 18 males and 6 females in 24 cases of cHCC-CC.The age ranged from 36 to 68 years (mean age was 54.38).Tumour location:right hepatic lobe in 15 cases,left hepatic lobe in 6 cases,both left lobe and right hepatic lobe in 1 case,hepatic caudate and left lateral lobe in 1 case,diffuse nodular liver tumors in 1 case.Grossly,the texture and color of tumor was related to the composition of tumor.Microscopically,classic cHCC-CC had two areas composed of hepatocellular carcinoma area and cholangiocar cinoma area of mixed distribution or migration distribution.3 cases were cHCC-CC with stem cell properties (cholangiolocellular carcinoma type,CLC type).Immunohistochemical staining revealed that HCC like area mainly expressed CD10,CK8,Hepatocyte and CD10,cHCC-CC area expressed CK7 and EMA.CLC type expressed the immunophenotypic features of intermediate type of HCC.Conclusion The clinical manifestations of cHCC-CC are not specific,the preoperative diagnosis is difficult,and it should be combined with pathological characteristics,imaging features and immunophenotype diagnosis.