Background The pathogenesis of dry eye is complicated,hormone level is thought to be one of impact factors in the development of dry eye.The regulation of the synthesis process of metalloproteinases(MMPs) in tissue has been reported.However,the effects of hormone on expression of MMP-2 in lachrymal gland is not clear.Objective This study was to investigate the effects of estrogen and androgen on the expression of MMP-2 in lachrymal gland in ovariectomized rats,and explore the role of MMP-2 in dry eye.Methods Sixty-four 3-monthold clean female Wistar rats were randomized into control group(8 rats),sham operation group(8 rats)and experiment group(48 rats).Ovariectomy(OVX) was performed on the rats of experiment group,and only fat tissue of abdominal cavity was cut off in the rats of the sham operation group.After 5 months of OVX,the experimental rats were subdivided into model control group,vehicle group,estrogen and androgen systemic or topical utilization groups and 8 rats for each group.Six weeks after administration of the drugs,the lachrymal gland was obtained.The expression of MMP-2 mRNA in the lachrymal gland was detected by reverse transcription PCR(RT-PCR),β-actin mRNA was used as an internal control,and the expression of MMP-2 protein was detected by Western blot,GAPDH was used as protein loading control.The use and care of the rats complied with the ARVO Statement.Results The expression of MMP-2 mRNA was strongest in the systemic estrogen group and was weakest in the systemic androgen utilization group.A significant difference in the MMP-2 mRNA expression was found among the 8 groups(F=18.60,P＜0.01),and the MMP-2 mRNA was significantly higher in the model group than that of the normal control group(0.66±0.10vs.0.47±0.10)(q=3.01,P＜0.05).In addition,the MMP-2 mRNA was significantly higher in the systemic estrogen group compared with the model group (0.83 ±0.10 vs.0.66-0.10) (q =2.79,P＜0.05) ; while the expression of MMP-2 mRNA was significantly declined in the systemic androgen group in comparison with the model group(0.12±0.04 vs.0.66±0.10)(q=11.41,P＜0.01).The MMP-2 protein presented with a strongest expression in the systemic estrogen utilization group and a weakest expression in the systemic androgen utilization groups.The expression level of the MMP-2 protein in the lachrymal gland was significantly different among the 8 groups(F =7.28,P＜0.01).The MMP-2 in the model group was significantly higher than that of the normal group(0.55±0.13 vs.0.38±0.08) (q =2.39,P＜0.05),and that in the systemic estrogen group was increased in comparison with the model group(0.69±0.12 vs.0.55±0.13) (q =1.85,P＜0.05).However,the MMP-2 in the systemic androgen group was significantly lowed in comparison with the model group(0.27±0.07 vs.0.55±0.13) (q =4.32,P＜0.01).Conclusions Estrogen may up-regulate the expression of MMP-2 in lachrymal gland,but the effect of androgen is opposite.Hormone level may play an important role in the regulation of the function of lachrymal gland.

Acquired Immunodeficiency Syndrome ; immunology ; HIV Infections ; immunology ; Humans ; Medicine, Chinese Traditional ; T-Lymphocytes ; physiology

To evaluate the value of silicone intubation and ring-fixed method in canalicular laceration.
●METHODS: A retrospective study of 36 cases with laceration of lacrimal canaliculus. For all the patients, microscope was used to find the broken ends of the lacrimal canaliculus, and then straight insertion was performed to the distant and near broken ends through the upper and lower lacrimal points, then passed the dacryocyst to insert nasolacrimal canal and ended up in inferior nasal meatus. Thus, clinical data about forming annular support to treat laceration of lacrimal canaliculus was presented here.
● RESULTS: All 36 patients with traumatic canalicular laceration were anastomosed successfully and all patients healed without infection. The surgery went well, intubation was smoothly, all patients were followed - up for another 6mo after the stent was removed. The accidental fall off of the stent was not observed during the follow - up. During the follow - up 10 - 18mo after extubation, when the stent was removed, 32 cases (88. 9%) recovered to normal lacrimal drainage function;4 cases ( 11. 1%) remained epiphora with obstructed lacrimal passage. All cases were no lower eyelid ectropion.
●CONCLUSlON: For the patients with inferior canalicular laceration, the silicone intubation and ring-fixed method is effective, low irritation and less complication.

This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.
Bacillus subtilis ; metabolism ; Biotransformation ; Fermentation ; Flavanones ; metabolism ; Flavonoids ; metabolism ; Glucosides ; metabolism ; Soy Foods ; microbiology

BACKGROUNDIt is known that ultraviolet irradiation can affect cellular function through a number of signaling pathways. (-)-epigallocatechin-3-gallate (EGCG) is the major effective component in green tea and can offer protection from ultraviolet-induced damage. In this study, we investigated the protective mechanism of EGCG on human dermal fibroblasts damaged by ultraviolet A (UVA) in vitro.

METHODSTranscription factor Jun protein levels were measured by Western blot. Matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA were studied by reverse transcription-polymerase chain reaction (RT-PCR) analysis in conjunction with computer-assisted image analysis. MMP-1 and TIMP-1 proteins were quantified by enzyme-linked immunosorbent assay (ELISA).

RESULTSEGCG decreased transcription activity of Jun protein after induction by UVA. Both the mRNA and protein levels of MMP-1 were increased by UVA irradiation, while no significant changes were observed in TIMP-1 levels. The ratio of MMP-1 to TIMP-1 showed statistically significant differences compared with the control. EGCG decreased the ratio of MMP-1 to TIMP-1 by inhibiting UVA-induced MMP-1 expression (P < 0.05).

CONCLUSIONEGCG can protect human fibroblasts against UVA damage by downregulating the transcription activity of Jun protein and the expression of MMP-1. The ratio of MMP-1 to TIMP-1, rather than the levels of MMP-1 or TIMP-1 alone, may play a significant role in human skin photodamage.

Catechin ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Fibroblasts ; metabolism ; radiation effects ; Gene Expression Regulation ; drug effects ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; analysis ; RNA, Messenger ; analysis ; Radiation-Protective Agents ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Ultraviolet Rays

From original concept and literature of acupoint, the concept and clinical significance of ashi method is discussed, which clarifies that the essence of ashi method is to locate the acupoints by patients' sensitivity on force. The clinical application of heat-sensitive moxibustion has illustrated that positioning method of this therapy is based on the appearance of heat-sensitive moxibustion sensation. Although both types are based on patients' feeling, positioning method of heat-sensitive moxibustion stands on a new angle and uses a new method to locate acupoint. Therefore, it is believed that the positioning method of heat-sensitive moxibustion is the inheritance and development of ashi method.
Acupuncture Points ; China ; History, Ancient ; Humans ; Medicine in Literature ; Moxibustion ; history ; methods ; Sensation

Adolescent ; Child ; Collagen Type I ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Fibrillin-1 ; Fibrillins ; Heart Defects, Congenital ; metabolism ; pathology ; Humans ; Male ; Microfilament Proteins ; metabolism

OBJECTIVE Glioblastomas(GBM)are the most malignant brain tumors in humans and have a very poor prognosis. New therapeutics are urgently needed. Here, we reported 2-methoxy-6-acetyl-7-methyljuglone (MAM)-induced cell death in U87 and U251 glioma cancer cells. METHODS Cells were cultured and treated with MAM, the cell viability was determined by MTT assay and LDH assay. Intracellular reactive oxygen species (ROS) generation was observed by DCF fluorescence. The protein expression was determined by Western blotting. RESULTS MAM induced glioma cancer cell death without caspase activation. The cell death induced by MAM was attenuated by the pharmacological or genetic blockage of necroptosis signaling,including RIP1 inhibitor necrostatin-1s (Nec-1s)and siRNA-mediated gene silencing of RIP1 and RIP3,but was unaffected by caspase inhibitor z-vad-fmk or necrosis inhibitor 2-(1H-Indol-3-yl)-3-pentylamino-maleimide (IM54). MAM treated U87 and U251 glioma cancer cells induced RIP1/RIP3 complex formation, ROS level increased, ATP concentration decreased and loss of plasma membrane integrity, further confirmed this process was necroptosis.The essential role of ROS was confirmed by the protective effect of ROS scavenger NAC. Interestingly, MAM induced necroptosis both triggered by RIP1/RIP3 complex and ROS generation. Moreover, MAM induced necroptosis through cytosolic calcium (Ca2 +) accumulation and sustained c-Jun N-terminal kinase (JNK) activation. Both calcium chelator BAPTA-AM and JNK inhibitor SP600125 could attenuate cell death. Further, we found there exists a feedback loop between RIP1 and JNK activation.Finally,MAM induced necroptosis was inhibited by dicoumarol(a NQO1 inhibitor). Dicoumarol exposed glioma cancer cells were resistant to RIP1/RIP3 complex formation and ROS generation. MAM induced necroptosis was independent of MLKL. CONCLUSION MAM induced non-canonical necroptosis through the NQO1-dependent ROS and RIP1/RIP3 pathway.This study also provided new insights into the molecular regulation of necroptosis in human glioma cancer cells and a promising approach for GBM treatment.

The specimens of ductal carcinoma in situ (DCIS) with early invasion, and specimens collected by core needle biopsy (CNB) tend to contain limited amount of invasive component, so it is imperative to explore a new technique which can assess HER2 gene status accurately for the limited invasive cancer component in these specimens. Dual staining technique of combining immunohistochemistry (IHC) for myoepithelial cells and single or dual probe chromogenic in situ hybridization (CISH) for HER2 gene was performed on routinely processed paraffin sections from 20 cases diagnosed as having DCIS with invasive cancer. Among them, 10 had fluorescence in situ hybridization (FISH)-confirmed amplification of HER2 and 10 had FISH-confirmed non-amplification of HER2. We successfully detected HER2 genetic signals and myoepithelial IHC markers (SMM-HC or CK5/6) simultaneously on a single section in all 20 specimens. Myoepithelial markers and HER2 signals detected by dual staining assay were consistent with those by individual technique performed alone. HER2 gene amplification results determined by dual staining assay were 100% consistent with those of FISH. Dual staining technique which allows simultaneous detection of myoepithelial marker protein and cancerous HER2 gene is feasible, and it has potential to be used in clinical practice for effective determination of HER2 amplification in limited invasive component.
Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Chromogenic Compounds ; Female ; Gene Expression Profiling ; methods ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization, Fluorescence ; methods ; Neoplasm Invasiveness ; pathology ; physiopathology ; Receptor, ErbB-2 ; genetics ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity

This study was aimed to investigate the suitability of FVIII gene BclI (intron 18)and HindIII (intron 19) site polymorphism for genetic diagnosis of patients with hemophilia A (HA) and their families, and for detection of carriers. The FVIII gene bclI (intron 18) and HindIII (intron 19) site polymorphism on the X chromosome of 8 patients with HA and 45 families members were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), the pedigree of patients was drawn by means of PediDraw software online provided by China Genetic Counseling Network. The results indicated that combination detection of BclI and HindIII sites could provide the diagnosis information for 5 out of 8 HA families with diagnostic ratio of 62.57%, especially 2 HA families were accompanied by mutation of 2 sites. Besides, the definite diagnosis for 6 out of 11 suspicious carriers in 8 families could be made with diagnostic ratio of 54.5%. It is concluded that the combination detection of BclI and HindIII sites for analysis of HA patient family can elevate the diagnostic rate of HA patients and carriers.
DNA Mutational Analysis ; Factor VIII ; genetics ; Female ; Hemophilia A ; diagnosis ; genetics ; Humans ; Introns ; Male ; Pedigree ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic