[Serum metabonomics study on Cr (â ¥ ) subchronic exposure rats based on UPLC-Q-TOF-MS/MS platform].

Objective: To ananlyze the toxic effects and mechanisms of Cr (Ⅵ) subchronic exposure based on metabonomics techniques. Methods: Twenty-nine female Sprague-Dawley rats were randomly divided into control group, low dose group and high dose group, 10, 9, 10, respectively. The control group, low dose group and high dose group were treated with 0, 10, 50 mg/L Cr (Ⅵ) for 90 days respec tively. The serum samples of rats with different dose of Cr (Ⅵ) treatment were detected Using UPLC-Q-TOF-MS/MS technique and data was analyzed by PCA, PLS-DA and OPLS-DA to compare with metabolic profile in different Cr (Ⅵ) dose treatments. Pathway analysis was performed using MetaboAnalyst 4.0 software. Results: UPLC-Q- TOF-MS/MS has stable detection performance and reliable experimental data. The control group, low Cr (Ⅵ) and high Cr (Ⅵ ) metabolic profiles of rats serum differences was obviously, and there is significant difference of serum metabolic profile among rats treated with different dose of Cr (Ⅵ) . 18 differential metabolites were screened between Cr (Ⅵ) low dose group and control group, 23 differential metabolites between Cr (Ⅵ) high dose group and control group. Compared to control group, there were 13 differential metabolites in both Cr (Ⅵ) high dose group and Cr (Ⅵ ) low dose group, such as 3-Hydroxy-11Z-octadecenoylcarnitine, Anserine, Farnesyl pyrophosphate, Linoleoyl ethanolamid e, Linoleyl carnitine, Lithocholate 3-O-glucuronide, LysoPC [20∶2(11Z, 14Z) ], LysoPC[20∶3 (5Z, 8Z, 11Z) ], LysoPC[22∶2(13Z, 16Z) ], PG[16∶0/22∶5(7Z, 10Z, 13Z, 16Z, 19Z) ], PI[18∶1 (11Z) /20∶4(5Z, 8Z, 11Z, 14Z) ], PI[20∶3(5Z, 8Z, 11Z) /18∶0], Serotonin. These differential metabolites were related to Glycerophospholipid metabolism, Tryptophan metabolism, Pentose and glucuronate interconversions, Terpenoid backbone biosynthesis. Conclusion: Cr (Ⅵ) subchronic exposure could induce the significant difference of serum metabolic profile. The differential metabolites induced by Cr (Ⅵ) subchronic ex- posure were mainly related to amino acid and lipid metabolism.

The expression changes and correlation analysis of high mobility group box-1 and tissue factor in the serum of rats with sepsis.

OBJECTIVE: To investigate the expression changes of high mobility group box-1 (HMGB-1) and tissue factor (TF) and their correlation in the serum of sepsis rat models. MATERIALS AND METHODS: 30 rats were divided into the sham-operated group, 15 rats were in the control group. The cecal ligation and puncture method was used to make the animal model with abdominal infection induced by sepsis. There were 15 rats in the sepsis group among which they were divided into 3 subgroups at different time points after modeling (after 6 hours, 12 hours, 24 hours). Cardiac function indicators of the rats in each subgroup were monitored, including heart rate (HR), left ventricular end-diastolic pressure (LVEDP) and left ventricular developed pressure (LVDP), and enzyme-linked immunosorbent assay (ELISA) was used to test the changes of the expression levels of HMGB-1 and TF in the serum of the rats after 6 hours, 12 hours, 24 hours. Pearson correlation analysis was used to analyze the correlation between HMGB-1 and TF. RESULTS: HR and LVEDP of the rats in the sepsis group were significantly higher than those of the rats in the control group. The differences were statistically significant (p<0.050). LVDP of the rats in the sepsis group was markedly lower than that of the rats in the control group. The differences were statistically significant (p<0.050). The expressions of HMGB-1 and TF of the rats in the subgroups of the sepsis group were higher than those of the rats in the control group after 6 hours, 12 hours, 24 hours; the expression levels of HMGB-1 and TF of the rats with sepsis increased with time. The differences were statistically significant (p<0.050). When the expressions of HMGB-1 and TF of the rats in the sepsis group were compared with each other within the group the differences were significantly different (p<0.050). The expressions of HMGB-1 and TF in the subgroups at the 24th hour were significantly higher than those at the 6th hour. The differences were statistically significant (p<0.050). The differences of the expression of TF of the rats in the control group were not statistically significant (p>0.050). There was a significant positive correlation between HMGB-1 and TF of the rats in the sepsis group (r=0.772, p=0.002). CONCLUSIONS: The expression levels of HMGB-1 and TF of the rats with sepsis gradually increased with time, and the level of HMGB-1 was positively correlated with the level of TF.

Factors influencing the long-term results of autologous microvascular submandibular gland transplantation for severe dry eye disease.

We assessed long-term outcomes of autologous microvascular submandibular gland (SMG) transplantation for severe dry eye disease and investigated factors influencing long-term results. From August 1999 to January 2015, 185 patients (200 eyes) with severe dry eye received SMG transplantation. Subjective assessments and ophthalmologic evaluations were performed before and after transplantation. Follow-up results showed successful transplantation in 180 of 200 eyes (success rate: 90%), resulting in marked symptomatic relief of xerophthalmia. Surgery failed due to vascular thrombosis (15 glands) and duct obstruction (5 glands). Follow-up data were available for 163 eyes. Epiphora occurred in 98 (60.1%) eyes and was effectively managed by surgical reduction of graft, topical atropine gel and botulinum toxin injection. Wharton's duct obstruction occurred in 16 (10.6%) eyes and was treated by duct reconstruction. Subjective satisfaction was achieved in 143 (87.7%) eyes. Mean score of fluorescent staining reduced from 11.25±1.42 to 7.25±3.37. Postoperative best-corrected visual acuity improved in 85 (56.3%) eyes. Our clinical experience proved that SMG transplantation is effective and grants long-term improvement in severe dry eye. Secretory function of transplanted SMGs remains active and stable. Blood vessel thrombosis, Wharton's duct obstruction, and epiphora are primary factors influencing results.

Low expression of lncRNA MEG3 promotes the progression of oral squamous cell carcinoma by targeting miR-21.

OBJECTIVE: The aim of this study was to explore whether maternally expressed gene 3 (MEG3) could facilitate the proliferation and migration of oral squamous cell carcinoma (OSCC) cells by selectively binding to miR-21, thereby participating in the progression of OSCC. PATIENTS AND METHODS: The expression levels of MEG3 and miR-21 in OSCC tissues and normal control tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The effects of MEG3 and miR-21 on cell proliferation and migration were examined by cell counting kit-8 (CCK-8), transwell, and scratch assay, respectively. Meanwhile, cell cycle was detected using flow cytometry. The binding relationship between miR-21 and MEG3 was confirmed by dual luciferase assay. In addition, MEG3 and miR-21 were simultaneously knock-down to figure out whether MEG3 could regulate the proliferation and migration of OSCC cells through targeted binding to miR-21. RESULTS: QRT-PCR results indicated that MEG3 expression in OSCC tissues was remarkably lower than that of normal control tissues. However, the expression of miR-21 was significantly higher in OSCC tissues. Meanwhile, it was found that inhibiting MEG3 expression in OSCC cell lines could significantly promote cell proliferation and migration, while the simultaneous inhibition of miR-21 showed the opposite effect. Dual Luciferase assay results revealed that MEG3 could selectively bind to miR-21. In addition, we demonstrated that the knockdown of MEG3 in Tca-8113 and CAL-27 cells partially reversed the inhibitory effect of downregulated-miR-21 on cell proliferation and migration. These results further suggested that MEG3 might regulate OSCC cell proliferation via selectively binding to miR-21. CONCLUSIONS: Low expression of MEG3 can promote the proliferation and migration of OSCC cells through targeted binding to miR-21.

[Expression of PAI-2 mRNA in peripheral blood leucocytes and regulation by sGC activator in pulmonary hypertension].

OBJECTIVE: To investigate the mRNA expression level of plasminogen activator inhibitor-2 (PAI-2) in peripheral blood leucocytes and regulation by soluble guanylate cyclase (sGC) activator in pulmonary hypertension. METHODS: The human pulmonary arterial smooth muscle cells were treated with sGC activator Cinaciguat. The mRNA and protein expression levels of PAI-2 were detected with Real-time PCR and Western blot. The fresh blood samples of 8 patients with pulmonary arterial hypertension (PAH) (collected at the China-Japan Friendship Hospital from November 2014 to March 2015), 16 patients with chronic thromboembolic pulmonary hypertension (CTEPH) (collected at the China-Japan Friendship Hospital from November 2014 to March 2015), 24 age- and gender- matched healthy controls (collected at Beijing Hospital in March 2015) were treated with Cinaciguat for 8 hours. Then RNA of peripheral leukocytes was extracted and performed with reverse transcription and Real-time PCR to detect the mRNA level of PAI-2, which was compared between healthy controls and patients with pulmonary hypertension, before and after the treatment of Cinaciguat. At last, the correlation of PAI-2 mRNA level and the clinic severity of pulmonary hypertension were identified. RESULTS: The mRNA and protein expression levels of PAI-2 were promoted by Cinaciguat in human pulmonary arterial smooth muscle cells. The baseline mRNA level of PAI-2 in peripheral leukocytes was significantly lower in PAH patients compared to the healthy controls (0.201±0.152, 0.660±0.440, P=0.021). There was no significant difference in the mRNA expression level of PAI-2 between the CTEPH patients and controls (0.428±0.364, 0.769±0.682, P=0.152). After Cinaciguat treatment, the mRNA expression levels of PAI-2 were up-regulated in PAH patients and CTEPH patients (1.352±1.127, 1.203±1.008), there was no significant difference in the mRNA expression level of PAI-2 among the PAH patients, CTEPH patients and controls (P=0.130, P=0.534). The baseline mRNA level of PAI-2 was negative correlated with echocardiography-estimated systolic pulmonary arterial pressure (sPAP) (r=-0.744, P=0.034). CONCLUSIONS: The mRNA expression level of PAI-2 is significantly down-regulated in peripheral blood leucocytes and up-regulated by sGC activator in pulmonary hypertension. PAI-2 could be used as potential biomarker of pulmonary vascular remodeling in PAH.

[Association between OR2T3 gene and pulmonary arterial hypertension].

OBJECTIVE: To explore the pathogenic genes of pulmonary arterial hypertension (PAH) and validate the association between OR2T3 gene and PAH. METHODS: Whole exome sequencing was performed in four patients and one healthy person as control in two pulmonary arterial hypertension pedigree; patient-specific variations were screened by bioinformatics methods and comparison between groups. To further analyze the association between these variations and PAH, Sanger sequencing was used to analyze the genotype of patient-specific variations of 30 patients with idiopathic PAH, 90 healthy people and 30 patients with chronic thromboembolic pulmonary hypertension. RESULTS: The preliminary findings of whole exome sequencing were 57 variations may be associated with PAH; Among them, there were 6 AG heterozygotes due to OR2T3rs148748995 in the 30 idiopathic PAH patients, while no G allele carrier was found in other healthy people of two pulmonary arterial hypertension pedigree (AⅠ-1, AⅡ-3, BⅡ-1) and 90 normal control, and the difference was statistically significant (P<0.05). The variation also didn't exist in 30 chronic thromboembolic pulmonary hypertension patients. CONCLUSION: OR2T3 gene may be the pathogenic gene of PAH and OR2T3rs148748995 could have a role in the development of PAH.

Microvascular autologous submandibular gland transfer in severe cases of keratoconjunctivitis sicca.

The objective is to evaluate the technique of microvascular autologous submandibular gland transfer for the treatment of severe keratoconjunctivitis sicca. From August 1999 to April 2002, 38 patients with severe keratoconjunctivitis sicca were treated by autologous submandibular gland transfer to the temporal region of the skull. The related vessels were anastomosed to the superficial temporal artery and vein. When the vein was too small, venous bridging was applied. Prior to cutting off the gland, the facial artery was preserved and infused with heparin in normal saline after the gland had been freed to allow inspection of the blood oozing from the three veins. This would be helpful in the selection of a relevant vein for anastomosis. Wharton's duct was transplanted to the upper lateral conjunctiva fornix, and the gland was left denervated. Postoperative scintigraphy with Tc99m pertechnetate, follow-up studies, and management of complications were performed. The transplantations were successful in 33 cases, their symptoms of xerophthalmia disappeared. The discomfort resulting from bright light and wind was also relieved. These patients could stop applying artificial tears. In five patients the transplanted glands did not survive. Epiphora occurred in eight cases. They were successfully treated by reducing the size of the graft. Obliteration of Wharton's duct took place in two cases and was treated by the reconstruction of the duct or duct orifice. Microvascular autologous submandibular gland transfer is a lasting and effective solution for severe cases of keratoconjunctivitis sicca.

National epidemiological survey of blindness and low vision in China.

According to the National Sample Survey of Blindness and Low Vision, the prevalence of blindness in China was 0.43%, resulting chiefly from cataract (41.06%), corneal diseases (15.38%), trachoma (10.87%), and glaucoma (8.80%); and the prevalence of low vision in China was 0.58%, of which the main causes were cataract (49.83%), ametropia/amblyopia (14.98%), trachoma (9.55%), corneal diseases (8.48%), chorioretinal diseases (6.27%), etc. Among children under 14 years of age, the leading cause of blindness and low vision was heredity (48.46%). Among elderly of 60 years and over, the leading cause of blindness and low vision was cataract (73.13%).

Second-order neural nets for constrained optimization.

Analog neural nets for constrained optimization are proposed as an analogue of Newton's algorithm in numerical analysis. The neural model is globally stable and can converge to the constrained stationary points. Nonlinear neurons are introduced into the net, making it possible to solve optimization problems where the variables take discrete values, i.e., combinatorial optimization.