Objective To investigate the autophagy effect and the crosstalk with apoptosis by UV in A375 cells.Methods GFP-RFP-LC3 lentivirus were used to evaluate the effect of autophagy after being irradiated with UV of different doses (0、10、30、50 mJ/cm2).After being treated with 30 mJ/cm2 irradiation,the apoptosis rate of A375 with or without autophagy inducer was evaluated by annexin V-FITC/PI with flow cytometry.Western blot was used to distinguish the biomarker of autophagy (BECN,LC3) and apoptosis (Caspase 3,9).Results After being irradiated with 10 mJ/cm2 or 30 rnJ/cm2 UV,the autophagosome was observed at 6 h and rich at 9 h.However,the dots of autophagy had been abundant continually from 3 h with 50 mJ/cm2.The ability of inducing autophagy of UVB is stronger than UVA.UVA and UVB showed synergistic effect in autophagy with the dose of 30 mJ/cm2.The Caspase 3 and Caspase 9 proteins were downregulated after 30 mJ/cm2 UV irradiation with autophagy biomarkers increasing,whereas the apoptosis biomarkers were enriched with the inhibition of autophagy.Conclusions UV can induce autophagy with more significant effect of UVB.Autophagy paly protective role by delaying apoptosis after 30 mJ/cm2 UV irradiation in A375.

Objective Study on the relations between Rh hemolytic disease of the newborn (HDN) and the influencing factors of producing anti-D. Methods D antigens of 32 RhD-negative pregnant women and their newborns are determined by indirect antiglobulin test (IAT) and absorption/elution test. With polymerase chain reaction (PCR) and direct genomic DNA sequencing, we detect the RHD gene in weak D pregnant women identified serologically, and we analyzed the situation of fetomaternal hemorrhage (FMH) of the D-negative women with more than 2 gestations with flow cytometry. Results Among 32 pregnant women of RhD-negative detected by first test, there are 18 pregnant women with two and more pregnancy. In these 18 pregnant women, 3 cases are identified as D el phenotype, 1 case is designated as D category VI type III, the rest 14 cases are truly D-negative pregnant women. Among the truly D-negative multi-pregnant women, 2 produce anti-D in sera and 13 are detected fetal erythrocytes in their peripheral blood by flow cytometry. However there are no anti-D detected in sera of D-negative first-pregnant women. Conclusion No anti-D allo-immune response were observed in all first-time pregnant women. In multi-pregnant women, however, 14.3% produce anti-D and result in HDN of Rh.

OBJECTIVE: To investigate the effect of ultraviolet B( UVB) on autophagy and apoptosis in human epidermal melanoma A375 cells. METHODS: i) A375 cells at logarithmic growth phase were exposed to UVB at doses of 10. 0 and15. 0 m J/cm~2. Then cells were collected at time point of 3,6,9 and 12 hours after irradiation. The effect of UVB on cell autophagy was observed by monodansylcadaverine staining and the effect of UVB on cell apoptosis was observed by acridine orange/ethidium bromide staining. ii) A375 cells of 10. 0 m J/cm~2 group and 15. 0 m J/cm~2 group were exposed to corresponding dose of UVB irradiation. Then cells were collected at time point of 18,24,36 and 48 hours after irradiation,and cell survival rate was examined using CCK-8 assay. iii) A375 cells were irradiated with UVB at doses of 10. 0 and15. 0 m J/cm~2 and then cells were collected at time point of 3,6,9 and 12 hours after irradiation. After that,A375 cells were irradiated at doses of 2. 5,5. 0,7. 5,10. 0 and 15. 0 m J/cm~2 of UVB,then cells were collected at time point of 9 hours after irradiation. The expressions of B-lymphoblastoma-2( Bcl-2),Bcl-2 related X protein( Bax),Bcl-2 interacting protein( Beclin-1) and microtubule-associated protein 1 light chain 3( LC3) Ⅱ were detected by Western blotting. A375 cells with no UVB irradiation were set as the control( pseudo-irradiation) in each experiment. RESULTS: i) Both autophagy and apoptosis of A375 cells induced by UVB irradiation at doses of 10. 0 and 15. 0 m J/cm~2 increased with time after irradiation. The effect on autophagy decreased at 12 hours time point with 15. 0 m J/cm~2 UVB irradiation. ii) The cell viability increased with time after irradiation in the 10. 0 and 15. 0 m J/cm~2 groups( P < 0. 05). From 18-48 hours after irradiation,the cell viability of the 10. 0 and 15. 0 m J/cm~2 groups was lower than that of the control group( P < 0. 05).From 24-48 hours after irradiation,the cell viability of the 15. 0 m J/cm~2 group was lower than that of the 10. 0 m J/cm~2 group( P < 0. 05). iii) The relative expression of Beclin-1 and LC3 Ⅱ protein at the 10. 0 m J/cm~2 group increased with time after 0-12 hours irradiation( P < 0. 05). The above changes of the 15. 0 m J/cm~2 group were observed within 0 to 9 hours,and the above two autophagy-related proteins were significantly decreased at the 12 hours time point( P < 0. 05).The relative expression of Bcl-2 protein at the 10. 0 and 15. 0 m J/cm~2 groups decreased with increasing time from 3 to 12 hours after irradiation( P < 0. 05). The relative expression of Bax protein increased with time from 0 to 12 hours after irradiation( P < 0. 05). The relative expression of Beclin-1 and LC3 protein in cells at 0. 0-10. 0 m J/cm~2,and the relative expression of Bax protein in cells at 0. 0-15. 0 m J/cm~2 increased with increase of irradiation dose( P < 0. 05). The relative expression of Bcl-2 protein decreased with increase of irradiation dose at 5. 0-15. 0 m J/cm~2( P < 0. 05). CONCLUSION: Autophagy and apoptosis of A375 cells can be induced by UVB irradiation at doses of 10. 0 and 15. 0 m J/cm~2. Autophagy induced by UVB irradiation at 10. 0 m J/cm~2 partially resisted the induction of apoptosis by UVB and enhanced cell viability. 15. 0 m J/cm~2 UVB-induced autophagy was insufficient to exert the above-mentioned effects,and the induction of apoptosis was the dominant effect.

Objective Wnt signaling plays an important role in the development and progression of renal cell carcinoma (RCC).This study aimed to evaluate the effects of the Wnt signaling inhibitor 15-oxospiramilactone on the proliferation , migration, cell apoptosis, and cycles of the human RCC cell line 786-0, and to investigate the possible mechanisms of this small molecule acting on RCC in ivtro. Methods We treated 786-0 cells with DMSO ( blank control group ) and 15-oxospiramilactone at the concentrations of1.25μmol/L (low 15 -OSL), 2.5μmol/L (medium 15-OSL), and 5μmol/L (high 15-OSL), respectively, for 72 hours.Then we observed the changes in the proliferation and migration of the 786-0 cells by MTT and scratch-wound assay and determined their apopto-sis and cycles by Annexin V-FITC/PI assay and flow cytometry . Results 15-oxospiramilactone significantly inhibited the growth of the 7860-cells, with the IC 50of 1.088 μmol/L at 72 hours, and decreased their migration distance (P<0.05).After 36 hours of treatment, the apoptosis rates of the 786-0 cells in the low, medium, and high 15-OSL groups were (12.17 ±0.56), (18.54 ± 1.07), and (50.74 ±1.28) %, respectively, significantly increased as compared with (7.85 ±0.42) %in the blank control group (P<0.05), and in an obviously concentration-dependent manner.15-oxospiramilactone remarkably reduced the number of cells in the G0/G1 phase and increased that in the G 2/M phase (P<0.05). Conclusion 15-oxospiramilactone can significantly inhibit the pro -liferation and migration and induce the apoptosis of 786-0cells in vitro.It may be a potential anti-RCC agent.

OBJECTIVETo determine the frequency of RHD1227A allele in Chinese Hans.

METHODSFor a total of 890403 ethnic Han blood donors, the D antigen was determined with a saline method and indirect antiglobulin test. The RHD1227A allele and number and type of zygosity of RHD gene were determined with PCR sequence specific primer (PCR-SSP). Allelic frequency was calculated through statistics.

RESULTSIn total 2385 donors were found to be Rh-negative, 108 individuals were found to be weakly positive for D antigen (including weak D and partial D phenotypes). The remaining 887 910 individuals were Rh-positive. Among the Rh-negative individuals, 516 were found with RHD1227A. Among these, 467 were RHD1227A/d and 49 were RHD1227A/RHD1227A. Two of 108 D antigen weak-positive individuals were found as RHD1227A/RHD+. In addition, 8 of 1073 random Rh-positive samples were found to be RHD1227A/RHD+. The allele frequency of RHD1227A in the population was calculated as 0.004 036. The figure should be 0.006 682 if calculated based on the detected rate of the allele in Rh-negative individuals, and 0.007 884 if calculated based on the reported average phenotype rate of DEL in Rh-negative individuals.

CONCLUSIONBy taking main influencing factors such as the RHD zygosity, the rate of RHD1227A and DEL phenotype may be determined. The allele frequency of RHD1227A in Chinese Hans is between 0.004 036 and 0.007 884.

Adolescent ; Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Exons ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Rh-Hr Blood-Group System ; genetics ; Young Adult

Objective To study the effect of PM2.5 combined with UVB treatment on human keratinocytes HaCaT. Methods The PM2.5 in the air was collected in Guangzhou and the metal ingredients were analyzed. The cells were divided into four groups:negative control group(NC group),simple PM2.5 treatment group with the concentration of IC50(PM2.5 group),simple UVB irradiation group with the dose of 30 mJ/cm2(UVB group)and PM2.5 combined with UVB treatment group(combined treatment group). The effects of different treatments on cell viability were measured by MTT assay and those of different treatments on apoptosis were detected by flow cytometry. The expressions of PARP and LC3 protein were detected by Western blot. Results The metal components in PM2.5 samples included Ca ,Zn ,Ba ,Al ,Cu ,Pb ,etc. After the treatment of PM2.5 on HaCaT cells ,we concluded that the IC50 was about 300 μg/mL. The inhibitory effect on cell viability after 24 h in different groups showed significant difference (P < 0.001) and the viability of the combined treatment group was the lowest (P < 0.001). Flow cytometry analysis showed that compared with that of NC group,the apoptosis rate of PM2.5 group(P < 0.01),UVB group(P < 0.01)and the combined treatment group(P < 0.01)increased,but the apoptosis rate in the combined treatment group was higher than that of PM2.5 group(P < 0.05),but lower than that in UVB group(P < 0.01). Western blot showed that the level of LC3-Ⅱ and PARP in another three groups was higher than that of NC group;PARP in the combined treatment group was lower than that in UVB group and LC3-Ⅱ increased compared with that in PM2.5 and UVB group. Conclusion PM2.5 can increase the harm of UVB on HaCaT cells and the main mechanism may be through increasing autophagy rather than apop-tosis.

Objective:To analyze the prognostic risk factors of patients with traumatic pancreatitis (TP) and establish an early combined prediction of multiple indicators model for TP.Methods:Patients admitted to the ICU of the First Affiliated Hospital of Zhengzhou University from June 2017 to June 2022 were collected retrospectively. Based on their prognosis, the patients were divided into two groups: the good prognosis group and the poor prognosis group. The general data such as sex, age, underlying diseases, Glasgow Coma Scale (GCS), acute physiology and chronic health evaluationⅡ (APACHEⅡ), injury severity score (ISS), bedside index for severity in acute pancreatitis (BISAP), and clinical test indices such as blood routine, blood coagulation, blood gas analysis, and liver and kidney function at admission were compared between the two groups. Univariate analysis and multivariate logistic regression analysis were used to screen the early independent predictors of poor prognosis of TP, and the prediction model of TP was established by combining all of the independent indicators. The receiver operating characteristic (ROC) curve of each independent predictor and prediction model was drawn, and the area under the curve (AUC), sensitivity, specificity, and optimal cut-off value were calculated to examine the diagnostic impact of each independent predictor and the combined prediction model.Results:There were statistically significant differences in the complication rate of mental disorders, GCS, APACHE II, combined craniocerebral injury, combined chest injury, activated partial thromboplastin time, fibrin(pro)degradation products, lactate, aspartate aminotransferase, glomerular filtration rate, amylase, lipase, NT-proBNP, myoglobin, procalcitonin, ISS, and BISAP between the good and poor prognosis groups (all P<0.05). Multivariate logistic regression analysis showed that lactate ( OR=1.636, 95% CI: 1.046-2.559), lipase ( OR=1.005, 95% CI: 1.001-1.008), and ISS ( OR=1.161, 95% CI: 1.064-1.266) were independent risk factors influencing the prognosis of patients with TP. Based on the risk factors listed above, a prediction model was created: Logit P=-9.260+0.492×lactate+0.005×lipase+0.149×ISS, and the ROC curve was plotted. The AUC curve of the prediction model was 0.96 (95% CI: 0.91-1.00). Conclusions:Lactate, lipase, and ISS are early independent risk factors associated with the prognosis of TP. Their combined multi-indicator prediction model has an excellent clinical prediction effect, which can provide a clinical reference for early prediction and treatment of TP.

Objective:To investigate the application value of dual-source CT urography with stellar photon detectors in the diagnosis of gout.Methods:Forty patients who were diagnosed with gout according to American College of Rheumatology Guideline for the Diagnosis of Gout and received treatment between April 2018 and May 2020 were included in the observation group. Forty patients who were concurrently diagnosed with osteoarthritis and received treatment in the same hospital were included in the control group. All patients underwent dual-source CT urography with stellar photon detectors and corresponding biochemical index detection. Blood levels of uric acid, urea nitrogen, creatinine, total cholesterol, and triglyceride were compared between the observation and control groups.Results:Blood levels of uric acid, creatinine, urea nitrogen, total cholesterol, and triglyceride in the observation group were (519.38 ± 97.91) μmol/L, (110.21 ± 18.29) μmol/L, (12.21 ± 3.29) mmol/L, (6.49 ± 1.22) mmol/L, (3.45 ± 1.89) mmol/L, respectively, which were significantly higher than those in the control group (310.45 ± 61.40) μmol/L, (86.22 ± 13.12) μmol/L, (6.82 ± 1.75) mmol/L, (4.75 ± 0.56) mmol/L, (1.98 ± 0.85) mmol/L, respectively ( t = 11.43, 6.741, 9.148, 8.198, 4.486, all P < 0.05). Dual-source CT urography with stellar photon detectors revealed that urate crystals (color coded as green) were detected in 3 and 36 patients from the control and observation groups, respectively, with the detection rate of 7.5% (3/40) and 90% (36/40), respectively. There was significant difference in urate crystal detection rate between the observation and control groups ( χ2 = 24.993, P < 0.05). In the control group, no obvious destruction of bone, tendon and ligament were observed, urate deposition, total volume of (1.023 ± 0.83) cm 3, was found in feet and knee joint of a small number of patients. In the observation group, there were 30 patients with uric acid crystals and bone destruction in the metatarsophalangeal joint ( n = 6), distal tibia ( n = 7), distal fibula ( n = 3), proximal talus ( n = 4), proximal calcaneus ( n = 6), and wrist joint ( n = 4). There were 20 patients with ligament or tendon damage, involving deltoid ligament ( n = 2), Achilles tendon ( n = 10), and extensor and flexor tendon ( n = 53). Total volume of uric acid crystals was (32.22 ± 5.83) cm 3. The volume of uric acid crystals deposited in the hand, elbow, feet and knee was (8.00 ± 4.92) cm 3, (5.32 ± 2.75) cm 3, (36.00 ± 15.54) cm 3, and (13.31 ± 9.14) cm 3, respectively. Conclusion:Dual-source CT urography with stellar photon detectors has a high sensitivity in the diagnosis of gout, can accurately locate and quantify uric acid crystals and is of high application value in the diagnosis of gout.

Objective To quantitatively explore the influence of knife sharpness on forearm wounds in knife slash cases. Methods The finite element models of the upper limb and knives with 3 degrees of sharpness (with sharp blade, blunt blade, wide blade) were developed based on human CT images and prototype of slash knife. The slash by 3 kinds of knives on the forearm at velocity of 4 m/s and duration of 10 ms was simulated, so as to analyze changes in contact forces, wound dimensions and energy. Results During the slash by knives with sharp, blunt, wide blade, the blades reached the ulna at about 65, 85, 95 ms, respectively. The corresponding slash forces were 846, 1 064 and 1 865 N; the wound lengths were 135.64, 105.47 and 99.23 mm; the wound depths were 38.77, 27.81 and 18.74 mm. With the sharpness of blade decreasing, the wound formation was slowed, the length and depth decreased and the slash force increased. The model system for slash knife with sharp blade had obviously greater total energy and inner energy, but smaller kinetic energy, compared with slash knife with blunt blade and wide blade. Conclusions The method for quantitatively assessing wound formation in knife slash upon the forearm was developed. The research findings deepen the understanding of biomechanical mechanism of wound formation by knife slash, and provide new scientific means for forensic investigation and court trial of knife slash cases.