Background It is determined that riboflavin/ultraviolet A (UVA)-induced corneal collagen crosslinking is able to increase resistance of cornea against enzymatic digestion and has antimicrobial efficacy for various kinds of bacteria in vitro.However,its in vivo study is less now.Objective This study aimed to evaluate the efficacy of iontophoresis-mediated corneal collagen crosslinking combined with or without drugs for Staphylococcus aureus keratitis.Methods Bacterial keratitis models were induced by the interstromaly injection of Staphylococcus aureus suspension with concentration 2× 109/ml in the right eyes of 40 rabbits,and then the rabbits were randomly classified into the model group,gatifloxacin eye drops group,riboflavin/UVA corneal crosslinking group and drugs+ crosslinking group.The smearing of corneal surface was performed for the identification of bacteria 24 hours after injection.Iontophoresis-mediated riboflavin/UVA crosslinking was applied on the eyes of the riboflavin/UVA corneal crosslinking group and drugs+crosslinking group,and gatifloxacin eye drops was topically used 7 times per day on the eyes of the gatifloxacin eye drops group and drugs+crosslinking group.The corneal inflammation was examined and graded under the slit lamp biomicroscope before and after treatment.Ocular anterior segment optical coherence tomography(AS-OCT),corneal histopathology and ultrastructure were examined 14 days after treatment.The living environment of the experimental animals was maintained at 21 ℃ with a 12-hour light and dark cycle.Animals used in this study were treated in accordance with the Weifang Medical College Animal Experimentation Ethic Committee (AEEC) guidelines.The study protocol was approved by the AEEC.Results Corneal inflammation and ulcer were observed,but no significant difference was found in the inflammatory grade among the 4 groups 24 hours after injection (x2=0.293,P＞0.05).In the 14th day after injection,the corneal ulcer area was smaller and corneal edema was milder in the drugs+crosslinking group compared with the model group,gatifloxacin eye drops group and riboflavin/ UVA corneal crosslinking group,showing a significant difference in the inflammatory grade among them (x2 =38.710,P＜0.001).The cornea thickness values of ulcer zone were (428.1 ± 146.2) μm on the 14th postinjected day in the drugs+crosslinking group,which was evidently higher than those in the model group,gatifloxacin eye drops group and riboflavin/UVA corneal crosslinking group,with a significant difference among the 4 groups (F =8.310,P＜0.001).A lower degree of destruction of cornea collagen and less inflammatory cells were seen in the cornea tissue of the drugs+ crosslinking group by haematoxylin and eosin staining in comparison with other 3 groups,and normal keratocytes were much more in the drugs + crosslinking group than those in other treated groups.Conclusions Iontophoresismediated corneal collagen crosslinking can alleviate Staphylococcus aureus keratitis.The combination of crosslinking with drugs has a better effectiveness than the administration of gatifloxacin eye drops only or riboflavin/UVA corneal crosslinking only.

AIM: To establish an experimental model of high intraocular pressure in mice by laser photocoagulation and to prepare for future research.
METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure (IOP) to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain ( HE stain ) , cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling ( TUNEL ) was performed on the retinal sections to determine apoptosis rate.
RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk (P<0. 05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.
CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.

Yinchenzhufu decoction (YCZFD) is a classic formula for treating Yin Huang syndrome, which can improve liver injury caused by cholestasis. However, the mechanism of action of YCZFD still remains unclear. This article used network pharmacology, molecular docking, animal experiments, and molecular biology methods to explore the mechanism of YCZFD in treating liver injury caused by cholestasis. A mouse model of acute cholestasis induced by lithocholic acid was used to investigate the effects of YCZFD on liver injury. The experimental procedures described in this paper were reviewed and approved by the Ethical Committee at the Shanghai University of Traditional Chinese Medicine (approval NO. PZSHUTCM190823002). The results showed that YCZFD could reduce the levels of blood biochemical indicators and improve hepatocyte damage of cholestatic mice. Then, multiple databases were used to predict the corresponding targets of YCZFD active components on cholestatic liver injury. An intersection target protein-protein interaction (PPI) networks based on String database and Cytoscape software was used to demonstrate the possible core targets of YCZFD against cholestatic liver injury. The results indicated that core targets of YCZFD include tumor necrosis factor, interleukin-1β, non-receptor tyrosine kinase Src, interleukin-6, etc. GO (gene ontology) and KEGG (kyoto encyclopedia of genes and genomes) enrichment analysis indicated that YCZFD may regulate the tumor necrosis factor signaling pathway, nuclear factor-κB signaling pathway, bile secretion, and other related factors to ameliorate the cholestatic liver injury. AutoDockTools software was used to perform molecular docking verification on the core targets and components of YCZFD. To verify the results of network pharmacology, UPLC-MS/MS method was used to determine the effect of YCZFD on levels of bile acid profiles in mouse liver tissues. It was found that treatment with YCZFD significantly reduced the content of free bile acids, taurine bound bile acids, and total bile acids in the liver tissues of cholestatic mice. Then, results from real time PCR and Western blot also found that YCZFD can upregulate the expression of hepatic nuclear receptor farnesoid X receptor, metabolizing enzyme (UDP glucuronidase transferase 1a1), and efflux transporters (bile salt export pump, multidrug resistance-associated protein 2, multidrug resistance-associated protein 3, etc) in cholestasis mice, promote bile acid metabolism and excretion, and improve bile acid homeostasis. Moreover, YCZFD can also inhibit pyroptosis and inflammation by regulating NOD-like receptors 3 pathway, thereby inhibiting cholestatic liver injury.

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

Lambl's excrescences are filiform structures attached to the edges of cardiac valves and have been associated with stroke. Here we report two patients with Lambl's excrescences who developed migraine-like headaches followed by cerebral infarction. Their Lambl's excrescences were first identified by transesophageal echocardiography. One patient was given aspirin and another had surgery for debridement of excrescences. Their outcomes were good. The migraine-like headache might be the first symptom of cerebral ischemia and might be triggered by micro-emboli originated from Lambl's excrescences. Patients with Lambl's excrescences should be closely monitored and surgery should be considered in recurrent stroke cases.
Adult ; Headache ; diagnosis ; etiology ; Heart Valve Diseases ; complications ; diagnosis ; pathology ; Humans ; Male ; Middle Aged ; Stroke ; diagnosis ; etiology

The purpose of this paper is to establish big data platform for drug,linking the commonly used large databases and combining with examples to show the application of platform in pharmacokinetic/pharmacodynamic data,small molecular biological treatment target molecules,drugs,targets and adverse drug reactions.The drug big data platform is used to provide original clinical trial data for the pharmacokinetic/pharmacodynamic studies of breast cancer drug and provide data for other related researches.

Objective To observe the application of high-speed vitrectomy system in combined surgery of phacoemulsification and vitrectomy .Methods Eighty-one patients (86 eyes) aged over 60 years with degree Ⅱlens opacity and vitreous retinal disorders treated between October 2012 and March 2013 were enrolled .The pa-tients included 43 males (46 eyes) and 38 females (40 eyes), at a mean age of (72.1 ±10.5) years (60-83 years).All the patients received routine ophthalmological examination , A/B mode ultrasound, and intraocular lens degree testing .With a 23G minimally invasive incision , phacoemulsification and intraocular lens implanta-tion were performed after closed the intraocular perfusion .With the viscoelastic agent left in the anterior cham-ber, high-speed vitrectomy was conducted using Resight noncontact wide -angle lens system , finished by filling with silicone oil , C3 F8 , or intraocular infusion fluid .Visual acuity , fundus condition , and postoperative compli-cations of the patients were observed .Results The combined surgery was successfully performed in all the pa-tients, with a mean operation time of (48.0 ±12.6) minutes (35-65 minutes).Among the 86 eyes, vision was improved in 80 eyes (93.0%), unchanged in 5 eyes (5.8%), and deteriorated in 1 eye (1.2%).After the surgery , 10 eyes (11.6%) developed corneal edema , and 25 (29.1%) showed anterior chamber reaction , all mitigated by treatment after 1 week.No other complications were reported .Conclusions For the elderly pa-tients with degree II lens opacity and the need of vitrectomy , the high-speed vitrectomy combined with phacoemul-sification and intraocular lens implantation can rapidly improve eyesight and reduce the incidence of postoperative complications .

OBJECTIVETo investigate the clinicopathological characteristics, surgical treatment and prognosis of gastrointestinal stromal tumors(GIST).

METHODSThe clinicopathological data of 84 patients with GIST undergone resection between April 1997 and June 2008 were analyzed retrospectively, and the prognosis was evaluated.

RESULTSOut of 84 cases, 42 tumors located in stomach, 24 in small intestine, 18 in other sites. Tumor sizes ranged from 0.5 to 25 cm(average 5.6 cm). Positive rate of CD117 expression determined by immunohistochemical methods was 96.4%. Seventy-nine cases underwent complete tumor resection, while 5 cases received palliative resection or biopsy. Seventy-eight patients were followed up and their 1-, 3-, 5-year survival rates were 92.0%, 79.2%, 72.0% respectively. The Fletcher's classification of malignancy risk groups for GIST was related to the survival rates(P=0.001). The differences of survival rate among very low risk group, low risk group and high-risk group were significant(P=0.003, P=0.000).

CONCLUSIONSComplete tumor resection in the initial operation of GIST should be emphasized. The Fletcher's classification of malignancy risk groups for GIST is related to the survival rate. Extended surgical resection is required for GIST of higher malignancy risk.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrointestinal Stromal Tumors ; mortality ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate

OBJECTIVETo find a new and effective way for the transfection of adenovirus vectors encoding HSP70 cDNA, so as to provide another possible method in gene therapy against ischemia and cellular hypoxia after burn injury.

METHODSThe replicated defective adenovirus vectors encoding HSP70 cDNA were encapsulated. Its acid resistance and dissolution in intestinal fluid were tested in artificial gastric juice and intestinal fluid. The expression of HSP70 gene which was transfected by the microcapsules orally was detected by RT-PCR.

RESULTSThe encapsulated replicated defective adenovirus vectors were viable in vitro. They exhibited good resistant to acid (resolution ratio less than 10%) and dissolution in intestinal juice (resolution ratio higher than 50%). The HSP70 gene expression of the tested rats was significantly higher than control, but there was no difference in the quantity of HSP70 induced by sodium arsenite or adenovirus transfection through injection by vein.

CONCLUSIONThe encapsulation of adenovirus vectors can successfully keep the viability of the virus in vitro and protect the virus from harmful effect of acid and enzyme in the gastric juice. Its nice dissolution in intestinal juice should ensure its absorption by oral transfection. The expression of the HSP70 gene after oral intake of this preparation is as high as that with other traditional transfection methods. It is possible that in the future the encapsulated replication of defective adenovirus vectors encoding HSP70 cDNA can provide a safer, convenient and effective way for gene therapy for burn patients.

Adenoviridae ; genetics ; Animals ; Burns ; complications ; Capsules ; Genetic Therapy ; Genetic Vectors ; HSP70 Heat-Shock Proteins ; administration & dosage ; genetics ; Hypoxia ; etiology ; prevention & control ; Intestinal Mucosa ; metabolism ; Ischemia ; etiology ; prevention & control ; Male ; Myocardium ; metabolism ; Rats ; Rats, Wistar ; Transfection

OBJECTIVETo investigate the effects and prognosis of surgical treatment in primary gastrointestinal stromal tumors (GIST).

METHODSThe clinicopathological data of 73 patients with primary GIST underwent operation from April 1997 to December 2007 was retrospectively analyzed, and the prognosis was evaluated too.

RESULTSAmong the 73 cases, 68 cases received complete tumor resection, among which 12 cases underwent laparoscopic operation; while palliative resection and biopsy only were carried out in the other 5 cases. There was significant difference in survival rate between the two groups (P = 0.000). The 1-, 3-, 5-year survival rates of the 66 cases had been followed up was 91.0%, 78.2% and 74.1%, respectively. The malignancy risk grades of GIST was related to the survival rates on statistical analysis (P = 0.002). Significant differences were found in the survival rates between the patients with very low grade, low grade and high grade malignancy tumors (P = 0.012, 0.002).

CONCLUSIONSComplete tumor resection should be emphasized in primary GIST, and more attention should be paid to the initial surgical treatment. Extended surgical resection is required for tumors of higher malignancy risk. The indications of laparoscopic surgery in GIST should be selected with caution for tumor complete resection.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; surgery ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Analysis