Objective To investigate the effect of hyperoxia on growth of type Ⅱ alveolar epithelial cells (AECⅡ).MethodsLungs of fetal rats at 19 days of pregnancy were collected,and AEC Ⅱ was isolated and cultured by differential adherence method.Cells were randomly divided into air group and hyperoxia group.In air group,cells were cultured in 5％ CO2 incubator.And cells in hyperoxia group were cultured in 5％ CO2+95％ O2 incubator.The growth,activity,cell cycle,cell apoptosis of AEC Ⅱ were observed at 2,4,6 and 8 days of culture.The interaction between different time and groups were analyzed by ANOVA of factorial design.Comparison of means was done by two-sample independent t test and one-way analysis of variance.Bonferroni correction was used during the comparisons.Results(1) Cell growth situation:in hyperoxia group,cell number was decreased from2 hto 8 h [(7.29±0.43)×105/ml,(2.68±0.37)×105/ml,(0.23±0.10)×105/ml and (0.00±0.00) × 105/ml],and lower than those in air group [(10.41 ± 0.24) × 105/ml,(27.90±1.91) × 105/ml,(27.12±0.85) ×105/ml and (26.29±1.59) × 105/ml](t=10.992,38.912,94.166and 49.696,P=0.000 respectively). (2) Cell activity:the living cells ratio in hyperoxia group at 2 d[(79.00±0.71) ％],4 d [(52.80±1.14)％] and 6 d [(31.60±1.52)％] was lower than those [(97.00±0.71)％,(97.20±0.84)％ and (95.00±0.71)％] ir air group (t=31.213,70.519 and 84.722,P=0.000 respectively).(3) Cell cycle:the cell ratios of G1 phase and S phase in hyperoxia group at day 4 [(66.82±1.20) ％ and (27.31±1.16) ％] and day 6 [(70.22±1.27) ％ and (30.31±1.40) ％] were significantly higher than that at day 2 and that in air group (P＜0.05 respectively).(4) Cell apoptosis:in hyperoxia group,the cell ratio of Annexin-V+/PI- subgroup at 4 h was the highest [(23.89 ± 0.52)％],followed by those at day 2 and 6 [(21.32 ± 0.43)％ and (1.47 ±0.61)％].While the cell ratio of Annexin-V+/PI+ was the highest at 6 h [(53.92± 1.64)％],followed by those at 4 h and 2 h [(45.03±1.01)％ and (12.17±0.60)％],which were all different with those in air group(P＜0.05 respectively).ConclusionsHyperoxia might inhibit cell activity and cell cycle of AEC Ⅱ and promote apoptosis.

Objective To investigate the effects of intestinal ischemia-reperfusion(I/R)on cerebral microglial activation in rats.Methods One hundred and twenty-eight healthy male SD rats weighing 250-300 g were randomly allocated to one of two groups(n =64 each):group sham operation(group S)and intestinal I/R group.Intestinal I/R was produced by occlusion of superior mesenteric stery for 90 main followed by reperfusion.Sixteen animals were sacrificed at each of the 4 time points:2,6,24 and 48 h of reperfusion in each group.Their intestines were obtained for microscopic examination.Their brains were harvested for detection of microglial activation (by immuno-histochemistry).The reactive oxygen species(ROS),MDA and NO contents and SOD,nitric oxide synthase(NOS)and inducible nitric oxide synthase(iNOS)activities in the brain were measured.Results The microglia were in quiescent condition.Ibal staining was negative or light in group S.Intestinal I/R significantly increased intestinal Chiu score,cerebral microglial activation at 6,24 and 48 h of repeffusion which peaked at 24 h of reperfusion in group I/R as compared with group S.Cerebral ROS,MDA,NO levels and NOS,iNOS activities were significantly higher while SOD activity was significantly lower in group I/R than in group S.Concluslon Intestinal I/R can activate microglia and induce the release of nitrogen and oxygen free radicals resulting in cerebral injury.

Objective To investigate the effects of hydrogen inhalation on the brain injury after intestinal ischemia/reperfusion (I/R) in rats.Methods Fifty-four healthy male Sprague-Dawley rats aged 6-8 months,weighing 285-350 g,were randomly allocated to one of 3 groups (n =18 each):sham operation group (group S),intestinal I/R group (group I/R) and hydrogen inhalation group (group H2).Intestinal I/R was produced by occlusion of the superior mesenteric artery for 90 min followed by reperfusion.2％ hydrogen was inhaled for 3 h starting from the end of ischemia.The cognitive function was detected at 1,2 and 5 days of reperfusion using Morris water maze test.The animals were sacrificed after the test and brains were isolated for detection of the cerebral edema and morphology in brain tissues.The cerebral water content ((wet weight-dry weight)/ wet weight × 100％) was measured.The pathological changes in the prefrontal cortex was observed under light microscope.The neuronal apoptosis was detected by TUNEL.Results Compared with the S group,the number of normal neurons in the prefrontal cortex was significantly decreased,the latency and swimming distance were both prolonged,the frequency of crossing the original platform was decreased,and the cerebral water content and the number of apoptotic neurons were increased in groups I/R and H2 (P ＜ 0.05).Compared with I/R group,the number of normal neurons in the prefrontal cortex was significantly increased,the latency and swimming distance were both shortened,the frequency of crossing the original platform was increased,the cerebral water content and the nunber of apoptotic neurons were decreased in group H2 (P ＜ 0.05).The pathological changes were obvious in I/R group,however,they were significantly attenuated in H2 group.Conclusion H2 inhalation can reduce the brain damage and improve the cognitive dysfunction after intestinal I/R in rats.

OBJECTIVETo study the relative expression of the genes involved in artemisinin biosynthesis in different tissues including roots, stems, leaves and flowers of Artemisia annua, and establish the relationship between gene expression and artemisinin accumulation, eventually leading to discover the mainly effective genes involved in artemisinin biosynthesis.

METHODThe 7 functional genes involved in artemisinin biosynthesis were detected at the level of expression by using qRT-PCR, and simultaneously the content of artemisinin in the 4 investigated tissues was detected in parallel.

RESULTThe 3 genes including HMGR, DXR and FPS which were involved in the upstream pathway of artemisinin biosynthesis showed the highest expression levels in flowers, and the 4 functional genes including ADS, CYP71AV1, CPR and AAR which were involved in the artemisinin-specific biosynthetic pathway were found to be expressed in all the 4 detected tissues. The highest expression level of ADS was found in leaves, then followed by flowers, and the lowest expression level of ADS was found in roots and stems. CYP71AV1 had highest expression level in flowers and lowest in leaves. CPR showed highest expression level in flowers, and AAR had lower expression levels in the other 3 artemisinin-specific pathway genes in all the tissues. The highest content of artemisinin was found in leaves (0.343 mg x g(-1)), then followed by flowers (0.152 mg x g(-1)), roots (0.062 mg x g(-1)) and stems (0.060 mg x g(-1)).

CONCLUSIONIn the biosynthesis of artemisinin, the upstream genes including HMGR from the MVA pathway, DXR from the MEP pathway and the checkpoint gene FPS were much more active in flowers, and this suggested that flowers might be the tissues of artemisinin precursor biosynthesis, and further DXR contributed more to artemisinin biosynthesis. The positive correlation of ADS expression and artemisinin content in tissues demonstrated that ADS played a very important role in artemisinin biosynthesis, which was the ideal target for engineering the artemisinin biosynthetic pathway. In summary, the functional genes involved in artemisinin biosynthesis do not express at the same level but synergistically.

Artemisia annua ; chemistry ; genetics ; metabolism ; Artemisinins ; metabolism ; Plant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction

Catharanthine content and agronomic traits in major Catharanthus roseus varieties were analyzed. It was found that there existed great difference in catharanthine content and agronomic traits among the varieties. Catharanthine content was the highest in variety Pacifica Polka Dot (PPD), reaching 3.79 mg g(-1) dry leaf weight, and the lowest in variety Cooler Pink (CP) with only 0.9 mg g(-1) dry leaf weight. Correlation existed in certain extent between catharanthine content and agronomic traits in C. roseus. Path analysis showed that among all the agronomic traits analyzed, internodal distance positively affected catharanthine content at significant level (P<0.05), with the path coefficient being 1.473. This study provides useful information for high-catharanthine content C. roseus introduction and breeding.
Catharanthus ; anatomy & histology ; chemistry ; metabolism ; Plant Leaves ; anatomy & histology ; chemistry ; metabolism ; Vinca Alkaloids ; analysis ; metabolism

Objective To study the changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and explore their correlation with delayed hemolytic transfusion reactions(DHTR).Methods Serum samples from children with thalassemia who received blood transfusion treatment from June 2022 to April 2023(ob-servation group)and healthy children who underwent physical examination(control group)in our hospital were collected.The levels of serum immunoglobulins(IgG subtype,IgM,IgA,IgE and IgD)were detected using flow cytometry CBA multi-factor quantitative detection technology,and the differences between the two groups were compared.The children were divided into 4 groups according to different transfusion numbers:≤10 numbers,11-30 numbers,31-50 numbers and＞50 numbers,and the differences between different blood transfusion numbers and serum immunoglobulin levels in each group were compared using one-way analysis of variance(ANOVA).Children with thalassemia with DHTR were in the hemolysis group,and children with thalassemia who did not experience DHTR were in the non-hemolysis group.The changes in serum immunoglobulins(IgG subtypes,IgM,IgA,IgE and IgD)between the two groups were compared to explore the correlation between serum immunoglobulins in thalassemia children with repeated transfusion and DHTR.Results The levels of IgG1,IgG3,IgG4 and IgA in the observation group were significantly higher than those in the control group,with the increase of(2.07±2.12),(0.67±2.03),(0.30±0.37)and(6.04±11.40)mg/mL,respectively,while the level of IgD in observation group was significantly lower than that in the control group,with a decrease of(0.03±0.01)mg/mL,P＜0.05.No significant difference was noticed in IgG2,IgM and IgE between the groups(P＞0.05).IgG1 and IgG4 both significantly increased with the number of blood transfusions.The IgG1 in the 4 groups increased sequentially as(0.30±0.62),(0.41±0.51)and(3.60±3.48)mg/mL,and IgG4 increased sequentially as(0.12±0.13),(0.22±0.07)and(0.21±0.38)mg/mL.IgG2,IgM and IgD showed a significant decrease,with IgG 2,IgM,and IgD in four groups decreased as(0.91±1.50),(0.14±0.10)and(0.05±0.05)mg/mL,respectively,showing significant differences with the number of blood transfusions(P＜0.05).No sig-nificant difference was found in IgG3,IgA and IgE with different number of transfusions(P＞0.05).IgG1,IgG3 and IgG4 in the hemolysis group were significantly higher than those in the non-hemolysis group,with an increase of(4.44±3.41),(0.73±1.26)and(0.52±0.40),respectively(P＜0.05).IgD in the hemolysis group was significantly lower than that in the non-hemolysis group,with a decrease of(0.00±0.06)mg/mL,P＜0.05.No significance was noticed in IgG2,IgM,IgA and IgE between the hemolysis group and the non-hemolysis group(P＞0.05).Conclusion The serum immunoglobulin levels of children with thalassemia who undergo repeated blood transfusions are abnormal.There are differences in correlation between the number of blood transfusions and serum immunoglobulin levels among children with thalassemia who undergo repeated blood transfusions.The relevant serum immunoglobulins for DHTR in children with thalassemia who undergo repeated blood transfusions are IgG1,IgG3 and IgG4.

【Objective】 To analyze the effect of intravenous immunoglobulin(IVIG)-produced IgG antibody on the crossmatch incompatibility of neonates. 【Methods】 Blood type grouping, antibody screening, crossmatch, direct anti-globulin test, elution test, indirect antiglobulin test, and IVIG titer determination were conducted by microcolumn gel method. 【Results】 IgG anti-A were detected out in the elution test and free antibody test of 6 infants, and the titer of IgG anti-A contained in IVIG was 256, which led to the crossmatch incompatibility between infants and donors with the same type. 【Conclusion】 The hemolysis and crossmatch incompatibility in newborns, born to ABO-compatible mothers, may occur due to the IVIG-induced IgG antibodies. The O-type washed red blood cells should be selected for transfusion.

Objective To investigate the mechanism of action of integrin α4 (ITGA4) in liver fibrosis based on the anti-liver fibrosis effect of sticky sugar amino acid (SSAA) in rats. Methods A rat model of liver fibrosis was induced by intraperitoneal injection of CCl 4 , and then colchicine and low-, middle-, and high-dose SSAA were used for intervention, with blank control group and SSAA group as control. After 12 weeks of experimental intervention, serum and liver samples were collected to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and HE staining and Sirius Red staining were used to observe the pathological conditions of liver tissue; quantitative real-time PCR was used to measure the transcriptional level of ITGA4, integrin β1 (ITGB1), transforming growth factor-β1 (TGFβ1), alpha-smooth muscle actin (α-SMA), and TIMP2 in liver tissue; Western blot was used to measure the relative protein expression levels of ITGA4, ITGB1, TGFβ1, α-SMA, MMP2, TIMP1, and TIMP2; immunohistochemistry was used to observe the protein expression of TGFβ1 and α-SMA. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results There were significant increases in AST and ALT in the CCl 4 model group, and intervention with colchicine or low-, middle-, and high-dose SSAA reduced the levels of AST and ALT, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). HE staining and Sirius Red staining showed disordered structure of hepatic lobules and an increase in collagen fibers in the CCl 4 model group, and the structure of hepatic lobules was improved after intervention with colchicine or low-, middle-, and high-dose SSAA. The CCl 4 model group had significantly higher transcriptional levels of ITGA4, TGFβ1, α-SMA, and TIMP2 than the other groups, and there were significant reductions in the transcriptional levels of each factor after intervention with colchicine or SSAA, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). The CCl 4 model group had significantly higher protein expression levels of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and a significantly lower protein expression level of MMP2 than the other groups, and intervention with colchicine or SSAA inhibited the expression of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and promoted the expression of MMP2. Immunohistochemistry showed that the CCl 4 model group had significantly higher expression levels of TGFβ1 and α-SMA than the other groups, which was inhibited by intervention with colchicine or SSAA. The high-dose SSAA group had the most significant effect in reducing aminotransferases, improving lobular structure, and inhibiting the protein expression of liver fibrosis factors. Conclusion The high expression of ITGA4 in the liver is associated with the development of liver fibrosis, which is consistent with the increases in the expression of TGFβ1 and α-SMA. Inhibiting the expression of ITGA4 can provide more therapeutic targets for liver fibrosis and expand the anti-liver fibrosis mechanism of SSAA.

【Objective】 To investigate the changes in cellular immunity (peripheral blood lymphocyte subsets) and humoral immunity (serum immunoglobulin and ferritin) status among children with thalassemia who received repeated transfusions in Yunnan. 【Methods】 Forty-six children with thalassemia who underwent repeated blood transfusions from January 2020 to October 2022 were selected as the observation group. Forty children with thalassemia who did not receive blood transfusion were included in control group 1, and 46 healthy children underwent physical examination were included in control group 2. The differences in lymphocyte subsets, serum immunoglobulin levels and ferritin concentrations were compared among the three groups. 【Results】 For lymphocyte subsets: CD3⁺, CD4⁺ and CD4⁺/CD8⁺ in the observation group was lower than the control group 1 and 2: 57.60±8.36 vs 64.57±7.56 vs 66.58±5.65, 33.16±5.67 vs 38.62±8.36 vs 38.62±6.41 and 1.49±0.09 vs 2.32±0.15 vs 2.13±0.16, respectively; CD16⁺ CD56⁺ in the observation group was lower than the control group 2: 11.21±5.06 vs 16.70±7.92; CD8⁺ in the observation group was higher than control group 1 and control group 2: 26.63± 1.75 vs 20.60±1.43 vs 18.92±0.84; CD19⁺ in the observation group was higher than the control group 2: 24.06±6.42 vs 19.67 ±8.42, P<0.05, but no significant difference was noticed between the two control groups(P>0.05). For serum immunoglobulin and ferritin: IgG and ferritin in the observation group were higher than control group 1 and control group 2: 10.59±3.88 vs 7.02±3.88 vs 5.58±1.98 and 2 037.37±1 377.59 vs 72.63±56.71 vs 59.48±33.88. IgA in the observation group was higher than the control group 2: 1.06±0.92 vs 0.39±0.32(P<0.05), but no significant difference was noticed between the two control groups (P>0.05). The difference of IgM and IgE between the three groups was not significant (P > 0. 05). 【Conclusion】 The proportion of lymphocyte subsets in thalassemia children with repeated blood transfusion was imbalanced,and the level of immunoglobulin in humoral immunity was abnormal.