Objective Studying the status quo and constraints for rural healthcare service in grassroot rural healthcare units,for policy recommendations.Methods Using data from the fourth healthcare service investigation,by means of quantitative interview and qualitative interview,for an investigative interview of 348 township hospitals and 251 village clinics in 31 provinces in China.Results Deployment percentage of primary heahhcare services at township hospitals level is 49.1% (28.0/57),and that for village clinics is 60.6%(5.4/9).Conclusions The key to upgrading rural healthcare service system at grassroots level is to deploy better diagnostic equipments,upgrade the diagnostic competence of grassroots healthcare personnel and build a continuous service system for primary healthcare service.

OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.

METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.

RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.

CONCLUSIONYHN could inhibit C. albicans biofilms in rats.

Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.

METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.

RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.

CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.

METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.

RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.

CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.

Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology

Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents ; pharmacology ; Biofilms ; drug effects ; Candida albicans ; genetics ; physiology ; ultrastructure ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; HSP90 Heat-Shock Proteins ; genetics ; Microscopy, Electron, Scanning ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVETo investigate the effect of Shenfu Injection (SF, ginesenoside and aconite alkaloid) on the apoptosis of intestinal mucosal epithelial cells during ischemia-reperfusion in rats and its potential mechanisms.

METHODSIschemia-reperfusion model was established in rats. Twenty-four rats were divided into 3 groups with 8 rats in each, eg, ischemia-reperfusion (I/R) group, SF-treated group, and control group. In both SF and I/R groups, the superior mesenteric artery was closed with forceps for 1 hour and then reperfused for 2 hours. Either SF (3 ml/kg, SF group) or normal saline (I/R and control groups) was injected intravenously and continuously for 5 ml/kg with a micropump before the superior mesenteric artery was closed. The superior mesenteric artery was not closed for animals in control group. The expression of casapse-3 and Fas, and the level of TNF-alpha and pathological changes of the ileal mucosal tissue were assayed.

RESULTS(1) The number of apoptosis cells increased obviously in I/R group and was significantly higher than that in SF and control groups (P<0.05). (2) The expression of caspase-3, Fas, and TNF-alpha was significantly higher in I/R group than SF and control groups (P<0.01); however, there was not significant difference in the expression of capase-3 between control group and SF group. There was a positive correlation between the expression of caspase-3, Fas, and TNF-alpha, and the number of apoptosis cells. (3) Under light microscope, intestinal mucosal impairment was found milder in SF group than I/R group (P<0.05).

CONCLUSIONSSF can depress the apoptosis of intestinal mucosal epithelial cells during ischemia-reperfusion by restraining the expression of TNF-alpha, Fas, caspase-3, and accordingly alleviate the ischemia and reperfusion injury of intestinal mucosal epithelial cells.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; pathology ; physiology ; Female ; Intestinal Mucosa ; pathology ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; physiopathology ; Tumor Necrosis Factor-alpha ; metabolism ; fas Receptor ; metabolism

Objective To establish and evaluate a gene microarray for determination katG mutations of Mycobacterium tuberculosis isolates associated with resistance to isoniazid(INH).Methods A panel of probes were designed and gene chips were prepared by dotting.Mycobacterium tuberculosis isolates resistance to 5 drugs was determined by proportional dilution methods.Amplicons of Mycobacterium tuberculosis isolates were detected by our chip and sequenced.Results The drug resistance rate of the isolates to at least one of the anti-tuberculosis drugs was 70.8%(97/137).45 strains out 137 Mycobacterium tuberculosis isolates was resistant to INH(32.8%).katG was successfully amplified from 100% of the susceptible strains and 88.9%(40/45)resistant strains.4 of 45 INH resistant isolates' katG were deleted.27 of 40(67.5%) katG has been detected to have katG 315 codon mutations.The mutations were 315 AAC(Asn,13/40), ACC(Thr,6/40),ACA(Thr,4/40),ATC(Ile,2/40),AGC(Arg,2/40).The mutation rate of katG analyzed by gene chips we prepared were identical to katG sequencing.Conclusion The gene microarray techniques we developed for determination of Mycobacterium tuberculosis resistance to INH are specific, sensitive and may be used as an alternative in clinical laboratory.

Objective To retrospectively analyze and conclude the characteristics of imaging appearances of solid psedopapillary tumor of pancreas.Methods Among 7 cases with pathologically proved solid psedopapillary tumor of pancreas,6 cases underwent CT examinations of upper abdomen preoperatively,and the rest One had MRI examination.The mean age of these 7 cases(all female)was 30.7 years(range,14—44 years).Results The tumors were usually quite large in the largest diameter ranged from 2.8 to 15.9cm(mean largest diameter,7.9 cm);Tumors were all well demarcated,and 5 of them were of capsule on CT or MR imaging.All tumors were well-encapsulated on pathologic specimens, except for the capsule of 1 tumor was partially invaded;In 6 cases underwent CT examination,scattered, punctate and linear calcification were noted in the capsule of 2 tumors and the rim of another one;Except for 1 tumor was almost solid,the other 6 tumors contained both solid and cystic components;Scattered sheets of high attenuation shown in the cystic or solid parts on CT imaging in several cases and the high signal intensity on T_1-weighted MR imaging signified the possibility of bleeding in tumors,which then was testified by pathologic evaluation.Conclusion The solid psedopapillary tumor of pancreas has comparatively characteristic clinical and imaging features.

Objective To investigate the diagnostic value of prenatal ultrasonography in the fetal hemivertebra. Methods The ultrasonographic findings of three fetuses with hemivertebra diagnosed in our hospital were reviewed and compared with those of postnatal ultrasonography,other image modalities,and autopsy. Results In all fetuses,a distortion of the spine was observed where only one half of the vertebra could be identified. The parents opted for termination of the pregnancy in one case and the deformity was confirmed by autopsy. The other two fetuses were delivered and in one fetus the diagnosis was confirmed by radiological assessment. Conclusions Hemivertebra can be diagnosed accurately by second-trimester ultrasonography. The prognosis is mostly favorable when no other anomalies are associated. Meticulous examination may disclose the lesion and help decide the fate of pregnancy.

Objective To determine the diagnostic value of two and three dimensional ultrasound in detecting fetal hand malformations. Methods In the retrospectively analysis,the severe fetal hand malformations detected by prenatal ultrasound during the recent three years in our hospital were classified according to the prenatal sonographic characteristics,family history,karyotype analysis and autopsy results,etc.Results Fourteen fetuses with hand-anomaly were detected during the 16-28th week of gestation,both hands were affected in 10 cases,with the same morphology bilaterally in 8 cases.Thirteen cases had other abnormal sonographic findings.They were detected and classified into three categories,Wrist deformity (9 cases),three fetuses were associated with total absence of radius and radial clubhand,and six fetuses had palmar deviated hands,with various etiologies including familial hereditary arthrogryposis multiplex congenita,distal type 1 (AMCD1),amniotic band syndrome,body stalk anomaly,trisomy18 and micromelia.Hand (figer) hypoplasia or aphasia (3 cases),one hand was absent in one fetus without associated anomaly,absence of five fingers with ipsilateral multicystic dysplastic kidney in the second fetus,and the third fetus had split hand/foot malformation (SHFM).Overlapping fingers (4 cases),three of them were trisomy18,and two fetuses had both wrist deformity and overlapping fingers.Conclusions Prenatal two and three dimensional ultrasound play an important role in detecting and diagnosing severe type of fetal hand malformations.