The role of serum metrics in anti- VEGF treatment for macular edema induced by retinal vein occlusion.

AIM: To evaluate association between pretreatment serum metrics and best corrected visual acuity ( BCVA) of patients with macular edema secondary to retinal vein occlusion and its subtypes after intravitreal ranibizumab or conbercept implant. METHODS: This prospective research included 201 patients(201 eyes) who were diagnosed with macular edema secondary to retinal vein occlusion at Heibei Eye Hospital between January 2020 and January 2021, who all received intravitreal anti- vascular endothelial growth factor treatment. Serum metrics were measured before the first treatment, and correlations between BCVA and each of four parameters- platelets, neutrophil- to- lymphocyte ratio(NLR), platelet- to- lymphocyte ratio(PLR) and monocyte- to- lymphocyte ratio(MLR)- were analyzed to identify predictors of effective intravitreal injection treatment outcomes. RESULTS: The mean platelets was significantly different in the effective and ineffective group for RVO-ME (273.02 ± 41.49 × 109/L,214.54 ± 44.08 × 109/L P < 0.01),BRVO-ME (269.43 ± 49.52 × 109/L,214.72 ± 40.42 × 109/L P < 0.01), and CRVO-ME (262.32 ± 32.41 × 109/L,209.27 ± 42 0.91 × 109/L P < 0.01). The cutoff value of the platelets was 266.500, the area under the curve was 0.857,and the sensitivity and specificity were 59.8% and 93.6%, respectively. The mean PLR was significantly different in the effective and ineffective group for RVO-ME (154.66 ± 49.60, 122.77± 44.63 P < 0.01),BRVO-ME (152.24 ± 54.99, 124.72 ± 41.46 P = 0.003), and CRVO-ME (152.06±44.23, 118.67 ± 41.80 P = 0.001). The cutoff value of the platelets was 126.734, the area under the curve was 0.699, and the sensitivity and specificity were 70.7% and 63.3%, respectively. There were no statistical differencies between the effective and ineffective group(RVO- ME and its subtypes) in NLR and MLR. CONCLUSION: Higher pretreatment platelets and PLR were associated with BCVA in patients with RVO- ME and its subtypes who were treated with anti- VEGF drugs. The platelets and PLR may be used as predictive and prognostic tools for effective intravitreal injection treatment outcomes.

[Expression and catalysis of glucokinase of Thermoanaerobacter tengcongensis at different temperatures].

According to analysis of proteomic profiling for Thermoanaerobacter tencongensis, TTE0090 could be a novel gene of glucokianse (GLK), though no GLK gene was annotated in the genomic data. With the methods of cloning and expression in vitro, the recombinant TTE0090 was successfully expressed and purified. The recombinant TTE0090 exhibited the catalysis of GLK, even at high temperatures. Detection of expression levels and catalysis of TTE0090 in vivo was furthermore carried out at different temperatures. The expression of TTE0090 was attenuated during the culture temperature elevated; however, the specific activity was positively correlated to temperature raised. This leads a possibility that the metabolic capacity of glycolysis in T. tencongensis is relatively constant at different temperatures. All the results herein demonstrate that TTE0090 is a novel gene of GLK. The studies on TTE0090 and its protein product, thus, may deepen our understanding of the adaptation mechanism of thermophilic bacteria living in harsh environment.

[Cloning, expression and characterization of the 6-phosphofructokinase from Thermoanaerobacter tengcongensis].

The enzyme of 6-phosphofructokinase (PFK) is a key element in glycolysis, widely distributed in most eukaryote and prokaryote. Although the gene TTE1816 from Thermoanaerobacter tengcongenisis was annotated as a PFK based upon genomic analysis, its catalytic properties have to be examined experimentally. The cells of T. tengcongenisis were cultured at optimal temperature followed by the separation of the bacterial proteins with two-dimensional electrophoresis (2-DE). These 2-DE spots located around the theoretical values of pI and MW for TTE1816 were picked up and identified by mass spectrometry, suggesting that TTE1816 indeed expressed at such high temperature. Furthermore, TTE1816 was cloned into an expression vector and expressed soluble protein in E. coli BL-21 strain. The kinetic data revealed that the recombinant TTE1816 exhibited the catalysis to phosphorylate fructose-6-phosphate (F-6-P). Not only converting F-6-P to F-1,6-BP, does TTE1816 also catalyze the phosphorylation of glucose, fructose, mannose and glucose-6-phosphate(G-6-P) with optimal temperature at 60 degrees C . Interestingly, TTE1816 is capable to catalyze the reverse reaction as a bisphosphatase for dephosphorylation of F-1,6-BP under the reaction conditions with high concentrations of enzyme as well as substrates. The data reported herein demonstrate that a new member of PFK family has been identified in T. tengcongenisis.

Potential antitumor agent from the endophytic fungus Pestalotiopsis photiniae induces apoptosis via the mitochondrial pathway in HeLa cells.

4-(3',3'-Dimethylallyloxy)-5-methyl-6-methoxy-phthalide (DMMP) has previously been isolated from the endophytic fungus Pestalotiopsis photiniae. Although the cytotoxic activities of DMMP have been reported, little is known concerning the molecular mechanism of its cytotoxic effect. In the present study, we investigated the effect of DMMP on the growth of several types of cancer cell lines and investigated the mechanism of its antiproliferative effect. DMMP caused the growth inhibition of human cancer lines HeLa, MCF7 and MDA-MB-231, but had little antiproliferative effect on MRC5 normal lung cells. DMMP also significantly caused cell cycle arrest in the G1 phase and upregulated the cyclin-dependent kinase inhibitor p27KIPI protein in the HeLa cells. Moreover DMMP was able to induce marked nuclear apoptotic morphology in HeLa cells. DMMP induced apoptosis and loss of mitochondrial membrane potential (ΔΨm) in the HeLa cells. Although the activated forms of caspase-9 and -3 in HeLa cells were detected, pretreatment with caspase inhibitors (Ac-DEVD-CHO and Z-VAD-FMK) failed to attenuate DMMP-induced cell death. In addition, protein levels of the p53 family members, p53 and p73, were upregulated, and DMMP significantly increased the mRNA expression of pro-apoptotic Bcl-2 family genes (PUMA, NOXA, Bax, Bad and Bim). HPV E6-E7 mRNA levels were reduced. In conclusion, DMMP demonstrates potential for use in the treatment of cervical cancer.

Molecular cloning and expression patterns of copper/zinc superoxide dismutase and manganese superoxide dismutase in Musca domestica.

Superoxide dismutases (SODs) are metalloenzymes that represent one important line of defense against reactive oxygen species (ROS). In this paper, two novel SOD genes, MdSOD1 and MdSOD2, which putatively encode 261 and 214 amino acid residues respectively were identified and characterized from the housefly Musca domestica. The high similarity of MdSOD1 and MdSOD2 with SODs from other organisms indicated that they should be two new members of the SOD family. qPCR exhibited a universal expression of MdSOD1 and MdSOD2 detected in various tissues of housefly larva, including the fat body, gut, hemocyte and epidermis. Expression profiling reveals that MdSOD1 and MdSOD2 can be induced significantly via not only heat shock and cadmium (Cd) stress but also Escherichia coli and Staphylococcus aureus challenge. The two genes were cloned into the prokaryotic expression vector pET-28a to obtain the fusion proteins rMdSOD1 and rMdSOD2. Between them, the activity of rMdSOD2 was found by visual assay methods. ESI-LC-MS/MS analysis showed that three peptide fragments of the protein rMdSOD2 were identical to the corresponding sequence of M. domestica MdSOD2. MdSOD1 and MdSOD2 in housefly larvae were abrogated by feeding bacteria expressing dsRNA. High mortalities were observed in the larvae treated with dsRNA of SODs at heat shock, Cd stress and bacterial invasion. This phenomenon indicated that MdSOD1 and MdSOD2 are related to the survival of M. domestica under stress. This may provide new insights into the role of the two SOD genes in protecting M. domestica against both stress and bacterial invasion.