[The influence of knocking down the expression of low-density lipoprotein receptor associated proteins on the vascular abnormalities in hepatocellular carcinoma and its mechanisms].

Objectives: To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms. Methods: Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT). Results: Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated (r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 (r=-0.167, P=0.044), the level of serum CEA (r=-0.061, P=0.032), and the level of serum ALT(r=-0.147,P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 (r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT (r=0.164, P=0.029). Conclusion: Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

Curcumin improves experimentally induced colitis in mice by regulating follicular helper T cells and follicular regulatory T cells by inhibiting interleukin-21.

To determine whether curcumin (Cur) can treat mice with experimentally-induced colitis by regulating follicular helper T cells (Tfh) and follicular regulatory T cells (Tfr) by inhibiting interleukin (IL)-21. In this study, 40 male C57BL/6 mice were randomly grouped into four groups, i.e., normal, trinitrobenzene sulfonic acid (TNBS), TNBS + curcumin, and TNBS + anti-IL-21. Mice with experimental colitis were induced by 100 mg/kg TNBS. The mice in the TNBS + Cur group were treated with 100 mg/kg curcumin for seven days, and mice in the TNBS + anti-IL-21 group were treated with anti-IL-21 (150 µg/mouse) once per week, intraperitoneally, starting on the second day after establishing the experimental colitis model. On day eight, the therapeutic effect of curcumin was evaluated by colon mucosa damage index (CMDI), histological examination, and disease activity index (DAI). Furthermore, the number of CD4 + CXCR5 + PD-1 + Tfh and CD4 + CXCR5 + FoxP3 + Tfr cells were measured by flow cytometry. The mRNA and protein expression of IL-21, Bcl-6, FOXP3, ICOS, and PD-1 in colonic mucosa was detected by reverse transcription polymerase chain reaction and the Western blot technique. Compared with the TNBS group, the DAI, CMDI, histological score, the number of CD4 + CXCR5 + PD-1 + Tfh cells, the expression of IL-21, Bcl-6, ICOS, and PD-1 were significantly decreased in the TNBS + curcumin group and TNBS + anti-IL-21 group; body weight, number of CD4 + CXCR5 + FoxP3 + Tfr cells, and the expression of FoxP3 were observably elevated in the TNBS + curcumin group (all P < 0.05). Curcumin may have a potential therapeutic effect on mice with colitis treated experimentally through regulation of the balance of Tfh and Tfr cells via inhibiting the synthesis of IL-21.

[Visual outcome of cataract surgery and its influencing factors in China].

Objective: To explore the visual outcome of cataract surgery and its influencing factors in China. Methods: A case-control study. The information of 2 078 479 patients who underwent cataract extraction of 31 provinces (municipalities and autonomous regions) recorded in the National Cataract Surgery Information Reporting System during January to December, 2017 were analyzed. The postoperative visual outcomes of 3 days after surgery and 3 months after surgery were analyzed. The effects of age, gender, preoperative corrected visual acuity, past medical history, and surgical methods on the results of cataract surgery were analyzed. Surgery effect was divided into ideal effect, moderate effect and poor effect according to the best corrected visual acuity after operation. Wilcoxon rank sum test and multinomial logistic regression model were used for statistical analysis. Results: There were 1 197 942 female patients (57.64%) and 880 537 male patients (42.36%) in all 2 078 479 cases. The average age was (69±11) years old. The main type of cataract was age-related (1 928 440 cases; 92.78%). There were 1 608 385 cases, 1 126 961 cases and 389 020 cases with complete corrected visual acuity before surgery, 3 days after surgery and 3 months after surgery, respectively. The median difference of corrected visual acuity of preoperative versus three days after surgery, and preoperative versus three months after surgery were -0.48, -0.52 respectively (after logarithm of the minimum angle of resolution conversion). The interquartile ranges were 0.60 and 0.73 respectively. The differences were statistically significant (Z=551 599.30, 206 815.35; both P<0.01). Multivariate analysis showed that postoperative complications (moderate effect: OR=0.50, 95%CI: 0.46 to 0.55; ideal effect: OR=0.31, 95%CI: 0.28 to 0.33), preoperative pupil abnormal (moderate effect: OR=0.52, 95%CI: 0.44 to 0.61; ideal effect: OR=0.55, 95%CI: 0.47 to 0.63), preoperative intraocular pressure abnormalities (moderate effect: OR=0.86, 95%CI: 0.79 to 0.93; ideal effect: OR=0.45, 95%CI: 0.42 to 0.49), history of hypertension (moderate effect: OR=0.90, 95%CI: 0.85 to 0.95; ideal effect: OR=0.88, 95%CI: 0.83 to 0.93) and history of diabetes (moderate effect: OR=0.86, 95%CI: 0.80 to 0.93; ideal effect: OR=0.92, 95%CI: 0.86 to 0.99) were predictors of worse outcome. Good preoperative corrected visual acuity (0.2 to 0.4 for moderate effect: OR=2.20, 95%CI: 2.06 to 2.34; for ideal effect: OR=5.25, 95%CI: 4.94 to 5.59; 0.5 to 0.7 for moderate effect: OR=1.08, 95%CI: 0.96 to 1.21; for ideal effect: OR=6.35, 95%CI: 5.69 to 7.08; above 0.8 for moderate effect: OR=0.73, 95%CI: 0.60 to 0.89; for ideal effect: OR=6.58, 95%CI: 5.51 to 7.86), small incision extracapsular extraction (moderate effect: OR=3.19, 95%CI: 2.35 to 4.33; ideal effect: OR=6.79, 95%CI: 5.13 to 8.97) and phacoemulsification (moderate effect: OR=2.12, 95%CI: 1.60 to 2.81; ideal effect: OR=3.34, 95%CI: 2.59 to 4.32) were predictors of better outcome. Conclusions: Visual acuity of patients in China improve significantly after cataract surgery. However, some patients still suffer moderate visual impairment. The effect of surgery can be affected by many factors, such as preoperative corrected visual acuity, preoperative intraocular pressure, preoperative pupil status, surgical approach and postoperative complications, etc. (Chin J Ophthalmol, 2021, 57: 63-70).

[Effect of different plasma treated zirconia on the adhensive behaviour of human gingival fibroblasts].

OBJECTIVE: To evaluate the effect of different cold atmospheric plasma (CAP) treatment on the surface chemical and physical properties of zirconia and adhensive behaviour of human gingival fibroblasts (HGFs) cultured on zirconia disks. METHODS: The zirconia disks were divided into four groups and treated using helium, argon and mixture of argon and oxygen cold atmospheric plasma for 90 s or left untreated. The surface morphology, wettability and chemical elements were analyzed right after treatment. Human gingival fibroblasts were grown from biopsies obtained from a periodontally healthy human subject during periodontal surgery. HGFs were seeded on zirconia disk, and cells density was measured at the time point of 3 hours. Indirect immunofluorescence (IIF) was performed for morphometric examination at the time point of 3 hours. RESULTS: The crystallographic structure of zirconia was analyzed previously and the results suggested that it fitted the properties of zirconium yttrium oxide. After helium, argon and mixture of argon and oxygen cold atmospheric plasma treatment, the surface morphology and roughness of zirconia disks remained the same. The contact angle of zirconia decreased significantly(P<0.05)after CAP treatment: from 68.38° to 17.90°. After different CAP plasmas treatment, the atomic percentage of carbon on the outermost surface of the three groups decreased, as did the surface C/O ratio. The surface C/O ratio of zirconia decreased from 1.07 to 0.33. Fibroblasts density increased on CAP treated disks, especially the ones treated by mixture of argon and oxygen CAP(P<0.05). Cells of the three CAP plasma treatment groups spread better and had more protrusions, as well as larger surficial areas. CONCLUSION: Based on the results of this study after being treated by different kinds of CAP plasmas for 90 s, the surface wettability increased and the elements changed significantly with no changes in the tomography and roughness of the materials. The CAP treatment enhances the adhensive behavior of fibroblasts on zirconia by increasing the oxygen functional groups and promoting the cell density. Wettability of zirconia, an important physicochemical property, has a vital influence on the cell behaviors.

[Analysis on ophthalmic human resource allocation and service delivery at county level in Mainland China in 2014].

Objective: To master the status of ophthalmic human resource allocation and service delivery at county level in Mainland China, and to provide the basis for strengthening the construction of grassroots ophthalmic personnel team and formulating feasible medium and long-term eye health plans. Methods: All the medical institutions that provided ophthalmic service at county level or above were investigated by online reporting. This study only included medical institutions at county level. Descriptive statistics were used. Results: Three thousand three hundred and fifty-nine medical institutions were included, of which 1 463 hospitals had independent departments of ophthalmology (43.55%). Each medical institution has 3.93 ophthalmologists, 4.59 ophthalmic nurses, 0.30 full-time optometrist and 0.21 full-time optician. Ophthalmologists are mainly undergraduate, and the ratio of senior, intermediate and junior titles is 1.0∶1.7∶1.9. The ophthalmic nurses are mainly graduated from junior colleges or secondary schools, and the ratio of senior, intermediate and junior titles is 1.0∶7.3∶22.3. There were statistically differences in the distribution of job titles of ophthalmologists (χ(2)=106.21, P<0.01) and nurses (χ(2)=128.54, P<0.01) in the eastern, central and western regions. Of each ophthalmologist, the annual number of outpatient, outpatient surgery, inpatient and inpatient operation was 2 377.62, 109.79, 124.03, and 85.97 respectively. Four thousand one hundred and sixty ophthalmologists were able to carry out cataract surgeries independently, accounting for 31.50% of all ophthalmologists. The quantity for annual per capita cataract operation among ophthalmologists who can perform cataract surgery independently was 181.81 per capita. The average beds rate of utilization was 68.91%, and that was less than 60% in 52.99% of medical institutions. Conclusions: Compared with 1998, ophthalmic health personnel at county level in Mainland China has expanded rapidly. But cataract ability of ophthalmologists and the number of cataract surgeries still need to be improved. The structure of ophthalmic nursing staff is irrational. The full-time optometrists and opticians are extremely scarce. It is necessary to cultivate high-quality ophthalmic health personnel and improve beds rate of utilization at county level. (Chin J Ophthalmol, 2018, 54:929-934).

[The determination of activated partial thromboplastin time, coagulate time and thrombin time in patients with epistaxis of indeterminate cause].

OBJECTIVE: To explore the coagulation mechanism of indeterminate epistaxis. METHOD: 36 cases with epistaxis of indeterminate cause were studies by mean of detecting activated partial thromboplastin time (APTT), coagulate time (CT) and thrombin time. The results of first APTT, APTT after 8 min and CT were observed. RESULT: 1. The first APTT in epistaxis and normal group didn't show statistical difference (P > 0.05). But the APTT after 8 min in epistaxis were significantly different compared with the first APTT and that in normal group (P < 0.01). 2. The CT prolonged in 33.3% patients with epistaxis, which was higher significantly than that in the normal group (P < 0.01). (3) The epistaxis thrombin time (TT) was longer than that in normal group. CONCLUSION: The result suggest that during blood coagulation in indeterminate epistaxis, the activated factors in internal coagulating system may decompose more quickly than that in normal group. Antiagglutinating factors increase in blood.