Associations Between Fundus Types and Clinical Manifestations in Patients with RDH12 Gene Mutations.

To study the associations between RDH12 gene mutations, fundus types, and clinical manifestations. In total, 46 patients with inherited eye diseases caused by RDH12 gene mutations were included in this study. High-throughput chip capture sequencing, Sanger sequencing, and gene panel detection were used to determine that RDH12 was the pathogenic gene. All patients underwent the following detailed ophthalmic examinations: visual acuity, visual field, intraocular pressure, fundus photography, electroretinography, and optical coherence tomography (OCT). Statistical analysis was used to evaluate the clinical phenotype. A total of 32 mutations were identified in 46 patients. The most common mutations were c.437T > A, c.184C > T, and c.524C > T; the corresponding amino acid changes were p.Val146Asp, p.Arg62Ter, and p.Ser175Leu. Of the 46 patients, retinitis pigmentosa (RP) was found in 31 (68.9%); leber congenital amaurosis (LVA) was found in 11 (24.4%); early onset of severe retinal dystrophy (EOSRD) was found in one (2.2%); cone rod dystrophy (CORD) was found in one (2.2%); and Stargardt disease was found in one (2.2%). There was a significant difference in best-corrected visual acuity among patients based on fundus type (p = 0.0124). Linear trend analysis showed that best-corrected visual acuity gradually decreased as the fundus type increased in severity. In addition, there was a significant difference in the incidence of night blindness among patients with different fundus types (p = 0.0429): types I and IV fundi were associated with the highest incidences of night blindness. RDH12 gene mutation can cause serious inherited retinal diseases, which primarily include RP and LCA. Combined with clinical symptoms and fundus types, the progression of the disease can be characterized and used to guide genetic diagnosis and gene therapy.

[Effect of polysaccharides from seeds of Vaccaria segetalis in alleviating urinary tract infection induced bladder injury by inhibiting NLRP3 inflammasome].

To study the mechanism of polysaccharides from seeds of Vaccaria segetalis( PSV) in the treatment of bacterial cystitis through the NLRP3 inflammasome pathway. The rat model of urinary tract infection was used and treated with PSV,and the urine and bladders were collected. The level of interleukin-10( IL-10) in rat urine was detected by enzyme linked immunosorbent assay( ELISA). Western blot and immunofluorescence staining were used to detect the expressions of sonic hedgehog( SHH) and NLRP3 inflammasome [NOD-like receptor thermoprotein domain 3( NLRP3),apoptosis associated speck like protein( ASC) and pro-caspase-1]. The expression of Toll-like receptor pathway was detected by RT-PCR. The death of 5637 cells induced by uropathogenic Escherichia coli( UPEC) and lactate dehydrogenase( LDH) release were evaluated using live/dead staining. The results showed that in the rat bladder,the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors were significantly up-regulated,and NLRP3 inflammasomes were significantly activated by UPEC infection. The administration with PSV could significantly increase the concentration of IL-10 in urine,inhibit the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors in bladder,and inhibit the activation of NLRP3 inflammasomes. A large number of 5637 cells were dead after UPEC infection and caused LDH production. PSV could significantly inhibit the death of 5637 cells and the release of LDH. In conclusion,PSV could inhibit the expression and activation of NLRP3 inflammasomes by inhibiting the Toll-like receptor pathway,thereby mitigating the bladder injury.

A novel cytotoxic flavonoid C-glycoside from premna fulva.

Premna fulva Craib, locally known as "Zhangu" in China, is a kind of traditional medicinal plant. A phytochemical investigation on this plant led to the isolation of a novel flavonoid glycoside along with three known analogues. The chemical structure of the new compound was determined by spectral and chemical analysis as apigenin 8-C-ß-D-xylopyranoside (1). Compound 1 showed weak inhibitory activities in vitro against to four tumor cell lines (HL-60, Bcap37, SMMC7721, and P388) with IC50 values of 12.58, 19.31, 31.02, and 48.19 µg/mL, respectively. The result might be helpful to explain the application of P. fulva in Traditional Chinese Medicine.[Figure: see text].

Aporphine Alkaloids from Illigera aromatica from Guangxi Province, China.

Three undescribed aporphine alkaloids laurodionine B (1), illigerine A (2), and N-formyl-laurolitsine (3) were isolated from the methanolic extracts of the Chinese medicinal plant, Illigera aromatica, together with three known analogues (4-6). The chemical structures of 1-6 were identified by spectroscopic methods including 1D and 2D NMR (1H, 13C, COSY, HSQC, and HMBC) and high resolution mass spectrometry (HRESIMS). Compounds 1-3 showed moderate inhibitory activities in vitro against two cultured tumor cell lines, Hela and SMMC7721, with IC50 values of 32.42-62.90 µM. Only compound 1 had in vitro cytotoxic activity against Bcap37 cells, with the IC50 value of 90.61 µM.

Effects of curcumin on interleukin-23 and interleukin-17 expression in rat retina after retinal ischemia-reperfusion injury.

OBJECTIVE: This study aimed to investigate the effect of curcumin on the retinal structure and the expressions of interleukin-23 (IL-23) and IL-17 in the rat retina after retinal ischemia-reperfusion injury (RIRI). METHODS: 150 Sprague-Dawley rats were randomly divided into RIRI group (MG), low-dose curcumin group (LDCG) and high-dose curcumin group (HDCG), (n = 50 per group). RIRI was generated by anterior chamber perfusion of normal saline to the right eye. The left eye served as a normal control group (NCG). Rats in LDCG and HDCG received an intraperitoneal injection of 20 mg/kg/d and 100 mg/kg/d curcumin respectively, at 30 min before RIRI and once daily after RIRI. RESULTS: The morphological changes in HDCG group were improved as compared to MG and LDCG groups. Immunohistochemistry showed that IL-23 and IL-17 were mainly expressed in the ganglion cell layer and the inner nuclear layer of the retina. Low IL-23 and IL-17 expressions were observed in NCG, but increased significantly in MG and LDCG groups. Western blot assay and ELISA also showed that IL-23 and IL-17 expressions increased significantly after RIRI (vs. NCG, P<0.01). Moreover, the IL-23 expression reached a peak at 24 h, whereas IL-17 expression peaked at 72 h after RIRI. Curcumin reduced IL-23 and IL-17 expressions significantly in a dose-dependent manner (vs. MG, P<0.01). CONCLUSION: The IL-23 and IL-17 expressions increase after RIRI and curcumin significantly reduces retinal IL-23 and IL-17 expressions in a dose-dependent manner and is able to prevent the RIRI induced damage to the retina.

Isolation of two new C-glucofuranosyl isoflavones from Pueraria lobata (Wild.) Ohwi with HPLC-MS guiding analysis.

A profiling analysis of the total isoflavone extract of the root of Pueraria lobata (Wild.) Ohwi was performed using HPLC coupled with ESI-ion trap mass spectrometry. A total of seven isoflavones were identified according to their retention times, UV, MS data, and comparing with the literature data. Among them, two proposed new compounds were isolated and their structures were determined to be 8-C-alpha-glucofuranosyl-7,4'-dihydroxyisoflavone and 8-C-beta-glucofuranosyl-7,4'-dihydroxyisoflavone, named as neopuerarin A (7) and neopuerarin B (6), on the basis of chemical and spectral analyses.

An HPLC/MS method for identifying major constituents in the hypocholesterolemic extracts of Chinese medicine formula 'Xue-Fu-Zhu-Yu decoction'.

In this paper, HPLC-ESI-MS technique was used to analyze the hypocholesterolemic extracts of 'Xue-Fu-Zhu-Yu decoction', a traditional Chinese medicine consisting of six crude drugs (i.e. Paeonia lactiflora, Ligusticum chuanxiong, Citrus aurantium, Carthamus tinctorius, Prunus persica and Bupleurum falcatum). A total of 17 compounds were identified and their plant derivations were also distinguished. Nine compounds among them were unambiguously determined as oxypaeoniflorin, amygdalin, albiflorin, paeoniflorin, ferulic acid, naringin, hesperidin, senkyunolide I and neohesperidin by comparing the retention times (tR), UV spectra and m/z values with those of the reference compounds. The other eight compounds were tentatively identified as prunasin, 6-hydroxy-kaempferol 3,6-diglucoside, 6-hydroxykaempferol 3-rutinoside-6-glucoside, galloylpaeoniflorin, 6-hydroxy-kaempferol 3-glucoside, anhydrosafflor yellow B, narirutin and kaempferol 3-rutinoside by MS2 spectra and the comparison of their UV spectra and MS spectra with literature data.

[Effects of Gardenia-aweto compound by different extraction method on antagonizing acute respiratory distress syndrome].

OBJECTIVE: To study the effect of Gardenia-Aweto compound (GAC) and two component on preventing acute respiratory distress syndrome (ARDS) by the rabbit model of ARDS induced by intravenous injection of oleic acid. To detect the efficiency component of GAC in preventing ARDS. METHOD: GAC was divided into two compounts, ethanol-soluble components (ESC) and ethanol-deposition components (EDC), based on polarity. Forty-three new zealand rabbits were randomly divided into five groups, the blank control group, the model group, the GAC groups, the ESC group, and the EDC group. The ARDS model was induced by intravenous injection of oleic acid. Dynamic changes of arterial blood gas, lung index, albumin in bronchoalveolar lavage fluid (BALF) in different groups and lung histological changes were observed and compared. RESULT: As compared with the blank group, in the model group, GAC group, ESC group, EDC group the arterial PO2 and oxygen saturation deprived continuously. While SO2 in GAC group at time points 30, 60, 90, 120 min (P < 0.05 or 0.01) and SO2 in ESC group at time points 30, 60, 90 min were higher than those in ARDS group. PO2 in ESC group at time points 30, 60 min (P < 0.05) were higher than those in ARDS group. The value of LI and W/D were higher in ARDS group than in sham group (P < 0.01), they were much lower in HD group than in ARDS group (P < 0.01). Concentration of BALF-albumin increased markedly in ARDS group and pre-treatment groups compared with sham group, but it was much lower in GAC group and ESC group, there was a significant difference between GAC group (P < 0.01), ESC group (P < 0.05) and ARDS group. The lung histological changes had been improved in GAC group and ESC group. But no significantly difference between above-mentioned parameters was found in comparison in the model group and in the EDC group. CONCLUSION: Preventive administration of GAC or ESC an protect the damaged lung function in ARDS rabbits induced by oleic acid. The efficiency component of GAC in preventing ARDS is ESC. GAC antagonizing ARDS may relate to its anti-inflammatory, immuno-modulatory, anti-oxidant and antithrombotic effects.

[Discovery of the main active constituents of siwu mixture for reversing the multidrug resistance of human erythrocyte leukemic cell line K562/ADM].

OBJECTIVE: To study the main active constituents of Siwu mixture for reversing the multidrug resistance (MDR) of human erythrocyte leukemic cell line K562/ADM. METHOD: Macroporous resin was used to separate Siwu mixture and an in vitro model was used to evaluate the reversal index of each fraction for discovery of active fraction. HPLC-MS was used for investigating the main active constituents of Siwu mixture. RESULT: The reversal index of the gross polysaccharide of Rehmannia glutinosa was significantly higher than that of blank control (P < 0.01). There was little difference between the reversal effect of the gross polysaccharide of Rehmannia glutinosa and that of the active fraction A (P > 0.05). CONCLUSION: The gross polysaccharide of Rehmannia glutinosa is one of the main active constituents of Siwu mixture.

[Coordination inhibition of the proliferation of MCF-7 by enhancing expression of p53,p21 and decreasing expression of c-myc].

To study the coordination inhibition of the proliferation of breast cancer cell line MCF-7 by enhancing expression of p53,p21 and decreasing expression of c-myc, as well as the relationship among these genes when they express in the cells, eukaryotic expression plasmids of sense p53,p21 and antisense c-myc were constructed first, different concentrations of three plasmids were determined and the combinations were designed according to factorial design. Cells were transfected by the combination plasimids, then the inhibition rate of proliferation of the transfected cells were tested. The results were analyzed using the methods of statistics including Jin's Q-test, Least significant difference (LSD), cluster analysis. It was shown that all of them (Sense p21,p53, antisense c-myc) could inhibit the proliferation of MCF-7 in different levels of concentration, while the degrees of inhibition were different among them. In the aspect of coordination inhibition, the proliferation of MCF-7 was inhibited much strongly by p21 coordination with antisense c-myc and p53 coordination with antisense c-myc, but p53 coordination with p21 did not show inhibition effect. Rusults of cluster analysis showed that the first combination of three genes showed the best coordination, the ninth combination had the highest rate of inhibition. In conclusion, we suppose that when p53 as a cancer suppressor gene and p21 as a CDK suppressor gene express highly, and the expression of c-myc as a cancer gene is inhibited meantime could obviously enhance the inhibition on the proliferation of MCF-7 cells by coordination effect.