Background C57BL/6.NOD-Aec1Aec2 mouse is considered to be an idea model for the study of the pathogenesis of Sj(o)gren syndrome,but the cause of dry eye in these mice is unclear.Objective This study was to investigate the histopathological change of the ocular surfaces of C57BL/6.NOD-Aec1Aec2 mice,and to determine whether dry eye is developed spontaneously in these mice.Methods Forty-five clean C57BL/6.NOD-Aec1 Aec2 mice were used as the experiment group and forty-five C57BL/6J mice(both male and female)were used as the control group in this study.Detection of fasting blood-glucose,Schirmer' s test Ⅰ (S Ⅰ t),lissamine green staining and scoring of the corneal and conjunctival epithelium in the mice were performed at the age of 4,8,12,16 and 20weeks.Five mice from each group were sacrificed and their corneas were obtained to measure the central corneal epithelium thickness and to count the number of conjunctival goblet cells.In addition,lymphocyte infiltration in the lacrimal gland of the mice was examined with hematoxylin-eosin staining.The uhrastructure of the corneal epithelial cells and microvilli were assessed by scanning electron microscopy.The use and care of the mice were approved by the Experimental Animal Care Committee of the Third Military Medical University.Results No sign of dry eye was seen in both the 4-week-old C57BL/6.NOD-Aec1Aec2 mice and 4-week-old C57BL/6J mice.The S Ⅰ t values in 8-week-old,12-week-old,16-week-old and 20 week-old mice from the experiment group were (2.7 ±0.9) mm,(2.5 ±0.8) mm,(1.8±0.6) mm and (1.9± 0.1) mm,respectively,showing a significant reduction in comparison with those of the control mice of the same age(all P＜0.01).The amount of lissamine green staining in the C57BL/6.NOD-Aec1Aec2 mice gradually increased with age,showing elevated scores in 12-week-old,16-week-old and 20-week-old mice in the experiment group(all P＜0.01).The central corneal epithelium thicknesses were(20.18±3.75)μm,(17.01 ±5.25) μm,(14.19±5.72) μm and(12.00±3.25) μm in the 8-week-old,12-week-old,16-week-old and 20-week-old C57BL/6.NOD-Aec1Aec2 mice,respectively,which were significantly lower than those of the C57BL/6Jmice of the same age (all P＜0.01).The numbers of conjunctival goblet cells were (8.2±2.4),(6.2±2.1),(6.1 ±2.2) and (4.1 ± 2.0) in the 8-week-old,12-week-old,16-week-old and 20-week-old C57 BL/6.NOD-Aec 1Aec2 mice,respectively,showing a gradual decrease with age and a significant decline in comparison with those of the C57BL/6Jmice of the same age(all P＜0.01).Lymphocyte infiltration in the lacrimal gland and destruction of gland ducts were seen by hematoxylin-eosin staining,and acinar abnormality aggravated with aging.Reduction of corneal epithelial cells and the number of microvilli were distinguished with aging under the scanning electron microscope.The fasting bloodglucose levels of the two groups were both less than 6.0 mmol/L,and no significant difference was found between them at any age(P=0.637,0.610,0.163,0.086,0.938).Conclusions C57BL/6.NOD-Aec1Aec2 mice develop dry eye spontaneously with aging.The course of disease and characteristics of dry eye in C57BL/6.NOD-Aec1Aec2mice is similar to human dry eye.The C57BL/6NOD-Aec1 Aec2 mouse is the perfect model to study the pathogenesis of dry eye.

Objective To comparatively analyze the difference and characteristics of high mobility group box-1 protein(HMGB1) level with the levels in the patients with different severities of acute biliary tract infection (ABTI) to provide reference for its clinical diagnosis and treatment .Methods One hundred cases of ABTI in our hospital were divided into the mild group (48 cases) ,moder-ate group (29 cases) and severe group (23 cases) according to the severity of the disease .The HMGB1 detection results were com-pared among 3 groups and the differences in different disease types ,sex and age were analyzed .Results (1)The HMGB1 level had statistically significant difference among 3 groups (P<0 .05) ,moreover the mild group< moderate group< severe group ;(2)in the detection results ,the HMGB1 level in the mild group and moderate group had no statistical difference between males and fe-males(P>0 .05) ,but in the severe group ,the HMGB1 level in males was significantly higher than that in females (P<0 .05);(3) the HMGB1 level in the mild group had no statistical difference among difference age groups (P> 0 .05) ,while in the moderate group and severe group ,the HMGB1 level in the patients aged > 60 years old was significantly higher than that in the patients aged ≤60 years old(P<0 .05);(4) in the above 3 groups ,the HMGB1 level in the patients with acute cholecystitis was signifi-cantly higher than that in the patients with acute cholangitis (P<0 .05) .Conclusion The study results analysis indicates that the severe the ABTI disease condition ,the serum HMGB1 level is also accordingly and relatively increased ,in the patients with different severity degrees of ABTI ,the serum HMGB1 level has significant differences in age ,sex and disease type ,which prompts that the HMGB1 level can be used as the laboratory index for predicting and reflecting the ABTI severity and can be paid attention to .

Objective To explore the effect of curcumine on the proliferation and heparanase expression in blood vessel endothelium cells isolated from ovarian cancer tissues. Methods Blood vessel endothelium cells were obtained from pathologically identified ovarian cancer tissues by primary culture. The control group was the normal blood vessel endothelium cells. The experimental groups was treated with curcumine at 2,10,50 and 250 ?g/ml for 24,48 h and 72 h respectively. The cytostasis rate and cell cycle changes of the primarily cultured cells were detected by MTT assay and flow cytometry respectively. The transcriptional and translation levels of heparanase were detected by RT-PCR and immunofluorescent staining. The MDA content and LDH activity in the supernatant were detected too. Results In the curcumine groups,the cell proliferation was inhibited in a dose-dependent and time-dependent manner ( P

Objective To observe the histopathologic and ultrastructural characteristics of cornea and adjacent conjunctiva in Mooren’s ulcer. Methods The samples of limbal and central cornea and adjacent bulbar conjunctiva taken from active Mooren’s ulcer after lamellar keratoplasty were cut into paraffin sections and ultrathin sections and observed by light and transmission electron microscopy. The samples taken from patients of a Terrien’s marginal degeneration and a bacterial corneal ulcer were used as controls. Results Chronic inflammation including lymphocytes and plasma cells infiltrating existed in bulbar conjunctiva and sclera of Mooren’s ulcer. Limbal corneal epithelium, Bowman’s membrane, anterior stroma and adjacent superficial sclera melted and the inferior stromal collagen disorganized. The epithelial basement membrane of ulcer progressive edge had been destroyed while the epithelium and stroma kept quiescent. Lymphocytes infiltrated in conjunctiva and corneal epithelium of Terrien’s marginal degeneration with normal epithelial basement membrane, while Bowman’s membrane was destroyed. The epithelial basement membrane of bacterial corneal ulcer was intact. Conclusion Bulbar conjunctiva may act as a local lymph node of Mooren’s ulcer. Epithelial basement membrane of Mooren’s ulcer may have some abnormality as it was invaded first during ulcer progressing and it’s valuable to have a further study.

The characteristic fingerprint of conventional dairy Nanhanshuishi was established by X-ray diffraction (XRD), based on similarity of caculation on public peaks by MATLAB software, and the feasibility of new dairy technology of microwave method was explored between XRD and the dissolution rate in artificial simulation gastric juices. The result showed that similarity of shared peak in XRD of conventional dairy Nanhanshuishi was > 95%, This XRD characteristic fingerprint of conventional dairy Nanhanshuishi had strong specificity, could be used to provide a reference for identification and quality evaluation. This study also showed that the similarity of microware dairy products and conventional dairy products was good, and the sample of microwave 15 min was the best, and new dairy method by the microwave could replace the traditional method.
Animals ; Chemistry, Pharmaceutical ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Medicine, Tibetan Traditional ; Microwaves ; Milk ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; X-Ray Diffraction

Genuine medicinal materials with special characteristics of Traditional Chinese Medicine (TCM), is recognized as high quality medicine. Both ancient records and modern research considered that the origin is an important reason for the formation of genuine medicinal materials. However, blindly transplanting of genuine medicinal materials has led to the quality decline and counterfeit medicines appeared in production or sale progress, which may increase the risk of accidents in TCM. Frequent accidents emerged in Chinese herbal affects its export. What's more, it is a great threat to the medication safety in TCM clinical. There is an urgent need to implement traceability systems of TCM, which could provide convenient information record and traceability of TCM circulation. This paper reviews a variety of technical methods for genuine medicinal materials traceability, and proposed the establishment of genuine medicinal materials traceability system based on two-dimensional code and network database.
Databases, Factual ; Drugs, Chinese Herbal ; chemistry ; economics ; standards ; Medicine, Chinese Traditional

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
Drugs, Chinese Herbal ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Plants, Medicinal ; genetics ; RNA ; Sequence Analysis, RNA ; Transcriptome

Objective To investigate the electrophysiological properties of layer Ⅱ/Ⅲ pyramidal neurons of the early postnatal stage rat visual cortex.Methods By using the whole cell patch-clamp recording technique combined with direct visualization of cells,we studied the passive and active electrophysiological properties as well as firing properties of layer Ⅱ/Ⅲ pyramidal neurons in acute rat visual cortical slices.Results Resting membrane potential,input resistance,membrane capacitance and membrane time constant of pyramidal neurons at postnatal days 11-13(P 11-13) were(-56.6?1.8)mV,(185.4?2.7)M?,(77.9?2.2)pF and(16.9?2.4)ms,respectively.Action potential amplitude and duration were(97.7?2.7)mV and(2.3?0.1)ms,respectively.Threshold potential was(-31.8?1.2)mV and afterdepolarizing potential was(-65.3?1.1)mV.When presented with long depolarizing current pulse of constant amplitude,most of the neurons exhibited pronounced adaptation of spike frequency.The steady-state firing frequency was(30.4?9.4)% of the first interval firing frequency.Conclusion The electrophysiological properties of layer Ⅱ/Ⅲ pyramidal neurons of the early postnatal stage rat visual cortex are not fully mature.Most of the neurons display regular firing patterns,but the degree of firing frequency adaptation is relatively small.

Background Intracameral or intracorneal administration of amphotericin B (AMB) can achieve significant therapeutic efficacy in the treatment of recalcitrant fungal keratitis in cases that do not respond to conventional antifungal therapy. However, the ocular pharmacokinetics of the two routes of administration is unclear.Objective The goal of this study was to investigate the level of amphotericin B in cornea and aqueous humor of rabbits after administration of AMB via three different routes. Methods Forty-five healthy domestic rabbits were randomly divided into three groups. 1% amphotericin B of 10 μg was intrastromally or intracamerally injected into 15 rabbits, respectively,in group A and group B. Topical 0. 25% amphotericin B was topically administered to the eyes with corneal epithelial debridement (group C). Experimental animals were sacrificed and the corneas and aqueous humor samples were obtained for the detection of levels of amphotericin B at 30 minutes,6 hours, 1 day,3 and 7 days by high-performance liquid chromatography (HPLC). Results Calibration curves were linear over the range of 0. 10-100. 00 mg/L. The concentration of 0. 10 mg/L was the lowest quantifiable limit. The recovery of amphotericin B ranged from 89. 1% -95.7% from aqueous humor samples and 81.4% -83.6% from the cornea samples. After a single injection,effective drug levels were achieved and maintained for 7 days in cornea in group A, exceeding the minimum inhibitory concentration at which 90% of isolates are inhibited (MIC90) for a wide spectrum of fungi and molds with significant differences in comparison with group B and group C ( P＜0. 05 ). Effective drug levels were achieved in the aqueous humor in group B at 30 minutes after a single injection, but drug levels decreased dramatically within 6 hours. The evident differences were found between group B and group A or group C (P＜ 0.05). A considerable amount of amphotericin B was detected in the cornea and aqueous humor in group C within 1 day.Conclusion Effective high drug levels can be reached in rabbit cornea and aqueous humor after intrastromal and intracameral injection, respectively. Penetration of topical amphotericin B was greatly elevated after epithelial debridement.

OBJECTIVETo study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution.

METHODSOverlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed.

RESULTSThe genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I.

CONCLUSIONThe nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.

Animals ; Cell Line ; China ; Cricetinae ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny