[Associations of parental thalassemia with preterm birth and low birth weight].

Objective: To investigate the association between the preterm birth and low birth weight and parental thalassemia. Methods: Pregnant women and their husbands receiving prenatal examination in local hospitals or maternal and child health centers in Jingxi and Debao in Guangxi from January to December 2017 were selected as study subjects. A total of 758 pregnant women with pregnancy outcomes and their husbands, who were both or alone diagnosed with thalassemia through thalassemia gene detection, were selected as case group and 758 pregnant women with pregnancy outcomes and their husbands, who were negative in thalassemia gene detection and hemoglobin electrophoresis test were selected as control groups. The case group were further divided into mother group, father group and both mother and farther group. Clinical and pregnancy outcome data of the study subjects were collected for the analysis on the association between parental thalassaemia and preterm birth or low birth weight by the independent sample t test, χ(2) test and Cox regression analysis. Results: The incidence of preterm birth in case group and control group was about 6.5% and 1.6% and the incidence of low birth weight in case group and control group was about 7.3% and 0.8%. After adjusting for possible confounding factors, Cox regression analysis results showed that mother suffering from thalassemia (aRR=3.45, 95%CI: 1.35-8.81, P=0.010), fathers suffering from thalassemia (aRR=4.93, 95%CI: 2.16-11.21, P<0.001) and both mother and farther suffering from thalassemia (aRR=5.13, 95%CI: 2.62-10.04, P<0.001) were associated with preterm birth. Mother suffering from thalassemia (aRR=12.98, 95%CI: 4.91-34.30, P<0.001), fathers suffering from thalassemia (aRR=9.40, 95%CI: 3.40-25.95, P<0.001) and both mother and farther suffering from thalassemia (aRR=10.74, 95%CI: 4.44-26.00, P<0.001) were associated with low birth weight. The newborn whose parent all suffered from thalassemia had higher risks for preterm birth (χ(2)=22.72, P<0.001)and low birth weight (χ(2)=34.03, P<0.001) compared with those only with mother or father suffering from thalassemia. Conclusion: Parental thalassaemia, including both sides and single side, might increase the risks of preterm birth and low birth weight for newborn, and the risks might be higher in newborn with both mother and father suffering from thalassaemia.

[Dose-response relationship between maternal hemoglobin concentration and preterm birth, in pregnant women].

Objective: To investigate the dose-response relationship between hemoglobin concentration and preterm birth, during pregnancy. Methods: With Zhuang ethnicity, a total of 12 780 pregnant women and their infants that admitted to Wuming、Pingguo、Jingxi、Debao、Longan and Tiandong hospitals, were recruited, in Guangxi Zhuang Autonomous Region, from January 2015 to December 2017. Non-conditional logistic regression method was used to analyze the effect of anemia on preterm birth during pregnancy. Dose-response relationship between hemoglobin concentration and preterm birth was explored, using the restrictive cubic spline model. Results: After excluding 2 053 pregnant women with hypertension or aged 35 years and over, results from the non-conditional logistic regression analysis showed that the risk of preterm birth in the anemia group was 1.29 times (OR=1.29, 95%CI: 1.04-1.59, P=0.019) of the non-anemia group in the first trimester. Data from the restricted cubic sample showed that there appeared nonlinear "L" dose-response relationship between hemoglobin concentration and preterm birth in the first trimester and "U" shape in the third trimester (non-linearity test P<0.001). Conclusion: There appeared nonlinear dose-response relationship between the hemoglobin concentration and preterm birth, both in the first and third trimesters.

[Bioinformatics on vascular invasion markers in hepatocellular carcinoma via Big-Data analysis].

Objective: To investigate the biomarkers in hepatocellular carcinoma and their prognostic value via GEO (Gene Expression Omnibus) and TCGA (The Cancer Genome Atlas) database. Methods: Datasets of hepatocellular carcinoma were downloaded from GEO (GSE67140) and TCGA. MicroRNA in SNU423, SNU449, HepG2, Hep3B, SNU398 cell lines which had low or high invasion capabilities were investigated and verified, in 81 patients with and 91 without vascular invasion hepatocellular carcinoma. The prognostic value of these microRNAs were studied via TCGA database,obtained from 362 patients with hepatocellular carcinoma, through Kaplan-Meier and Multivariate Cox proportional hazard analysis. Target genes were analyzed by GO and KEGG. Results: Expressions of hsa-mir-1180, hsa-mir-149, hsa-mir-744 and hsa-mir-940 were all up regulated in high invasion capable cell lines (SNU423, SNU449) and vascular invasion patients with hepatocellular carcinoma (logFC>1, P<0.05). Results from the Survival analysis showed that hsa-mir-1180 (HR=1.623, 95% CI: 1.114-2.365, P=0.012), hsa-mir-149 (HR=2.400, 95% CI: 1.639-3.514) and hsa-mir-940 (HR=1.704, 95%CI: 1.188-2.443, P=0.004) were independent risk factors on the prognosis of patients with hepatocellular carcinoma (P<0.05). The mechanism might be related to factors as immune response, focal adhesion and adherence junction signaling pathways. Conclusion: With TCGA and GEO data mining, we found that hsa-mir-1180, hsa-mir-149, hsa-mir-744 and hsa-mir-940 were all highly related to the prognosis of hepatocellular carcinoma, that enabled it to be used to further study the biomarkers related to the prognosis of hepatocellular carcinoma.

[Impact and changes of maternal hemoglobin on birth weight in pregnant women of Zhuang Nationality, in Guangxi].

Objective: To investigate the hemoglobin (Hb) levels during pregnancy and Hb changes from early pregnancy and association with birth weight on infants. Methods: Mothers of Zhuang Nationality who participated in the pregnancy care program and delivered at the Pingguo County Hospital from May 2013 to May 2015 were included in this study. Retrospective analysis was applied to collect data of health care and pregnancy outcomes. Multiple regression analysis and unconditional logistic regression model were used for data analysis. Results: The mean birth weight appeared as (313 5.92±435.84) grams. The Hb levels at early pregnancy showed significantly positive association with birth weight. Results from our study demonstrated that when Hb levels increased + 1 g/dl at early pregnancy, birth weight would increase 17.61(95% CI: 0.60-34.67) grams, in the adjusted model. The Hb levels at late pregnancy were significantly inversely associated with birth weight. Our findings suggested that when Hb levels increased + 1 g/dl at late pregnancy, birth weight would reduce 19.61(95% CI: -37.53 --1.70) grams in the adjusted model. Changes in Hb from early pregnancy stages were significantly inversely associated with birth weight after adjusting for confounders and Hb levels in the early pregnancy stages. The results also indicated that when Hb levels increased a + 1 g/dl from early to late pregnancy, the birth weight would decrease 32.63 g(95% CI: -48.93--16.32). Compared to the non-anemia group, the anemia group showed significantly increase of small-for-gestational-age (SGA)(OR=1.58, 95%CI: 1.08-2.32) in early pregnancy. Compared to women under the most reduction status, women with the least reduction had a significantly increase of SGA (OR= 1.87, 95% CI:1.24-2.81) among their infants. With the magnitude of reduction on Hb concentration during pregnancy, the risk of delivering babies with SGA showed a gradual trends of increase. Conclusion: Hb levels at early pregnancy were positively associated with birth weight, but the changes of Hb were inversely associated with birth weight at late pregnancy, in women of Zhuang Nationality. Anemia in early pregnancy and the low amplitude of decreased Hb concentration during pregnancy were both risk factors for newborns under less gestational ages.

[Outcomes of pregnancy among women with alpha-thalassemia minor: A retrospective study of Pingguo county in Guangxi Zhuang Autonomous Region].

Objective: To investigate the association between the value of α-thalassemia minor and the outcomes in pregnant women. Methods: A total of 445 pregnant women with α-thalassemia minor were selected as thalassemia group in the Pingguo County Maternal and Child Health Hospital of Guangxi from January 2011 to December 2015, with ratio of 1∶4 healthy pregnant women was randomly recruited as non-thalassemia group. Clinical characteristics and pregnancy outcomes of the two groups were retrospectively analyzed using methods including t test, χ(2) test, and logistic regression model and ROC curve. Results: There were no significant differences noticed in factors as age, BMI, gestational age and educational level of the two groups. Hemoglobin of the thalassemia group was significantly lower than that of the non-thalassemia group (P<0.001). Differences on parity, ethnicities or occupation were statistically significant. Results from univariate analysis showed that the proportions of low birth weight, small for date infant and 1 min Apgar score<7 were higher in the thalassemia group, but the ratio of adverse pregnancy outcomes was comparable on parameters as preterm birth, stillbirth, macrosomia. Findings from the unconditional logistic regression showed that pregnancy complicated with α-thalassemia minor appeared a risk for both newborns with low birth weight (aOR=2.29, 95%CI: 1.32-3.95) and small for date infant (aOR= 2.11, 95%CI: 1.16-3.84). The ROC curve showed that α-thalassemia minor combined with multiple indicators presented a certain predictive value on neonatal birth weight. Conclusion: Pregnancy complicated with α-thalassemia minor was likely to increase the risk of birth weight loss in newborns, suggesting that prenatal care for pregnant women with thalassemia be strengthened, in order to reduce the incidence of adverse pregnancy outcomes.

Meta-analysis demonstrates lack of a relationship between XRCC1-399 gene polymorphisms and susceptibility to hepatocellular carcinoma.

XRCC1-399 allele polymorphisms have been reported to be associated with susceptibility to hepatocellular carcinoma (HCC), but the conclusions of the various studies have been inconsistent. We conducted a meta-analysis of available studies to determine whether XRCC1-399 alleles influence susceptibility to hepatocellular carcinoma. We searched English-language databases, including PubMed, Medline and Embase, using terms such as "hepatocellular carcinoma" (or "HCC"), "X-ray repair cross-complementing group 1" (or "XRCC1") and "genetic polymorphism" (or "SNP"), among others; we also searched Chinese-language databases, including CNKI, VIP, Wanfang Data, and CBM, using terms such as "ganai", "ganxibaoai", "ganzhongliu", "duotaixing", and "X-xian xiufu jiaocha hubu jiyin 1". Eight independent studies, including 1604 HCC cases and 2185 controls, were included. The pooled odds ratio for XRCC1-399 was 0.99 (95% confidence interval = 0.75-1.31). We conclude that XRCC1- 399 gene polymorphisms are unrelated to risk for HCC.

Short communication: Pheromonicin-SA affects mRNA expression of toll-like receptors, cytokines, and lactoferrin by Staphylococcus aureus-infected bovine mammary epithelial cells.

Pheromonicin-SA (Ph-SA) is a newly developed, engineered multidomain peptide that has a bactericidal effect against Staphylococcus aureus. The objective of this study was to characterize innate immune responses by Staph. aureus-stimulated bovine mammary epithelial cells (BMEC) following treatment with Ph-SA. Primary BMEC from one lactating Holstein cow were isolated and exposed to Staph. aureus for 2 h, and then treated with rifampicin or Ph-SA. Total RNA was isolated from BMEC at 0, 2, 6, 12, and 24 h postinfection, and the mRNA expression of selected genes, including toll-like receptor (TLR)2 and TLR4, IL-1ß, IL-6, IL-8, tumor necrosis factor α (TNF-α), and lactoferrin, was quantified by real-time PCR. In the rifampicin group, increases in the expression of mRNA for TNF-α, IL-1ß, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection and in the expression of mRNA for TLR2 but not for TLR4 at 12 h postinfection. In the Ph-SA group, increases in the mRNA expression of TLR2, TNF-α, IL-1ß, IL-6, IL-8, and lactoferrin were observed at 6 h postinfection, and an increase in TLR4 mRNA expression was observed at 24 h postinfection. At 24 h postinfection, the mRNA expression of TLR4, TNF-α, IL-1ß, IL-6, IL-8, and lactoferrin was higher in the Ph-SA group than in the rifampicin group. In conclusion, Ph-SA might promote the expression of mRNA for TLR2, TLR4, the pro-inflammatory cytokines IL-1, IL-6, and TNF-α, the chemotactic factor IL-8, and lactoferrin in Staph. aureus-infected BMEC. Moreover, Ph-SA may be of value as an antibiotic in promoting innate immune responses by Staph. aureus-infected bovine mammary epithelial cells.

[Analysis of synovial membrane apoptosis in different arthropathy].

OBJECTIVE: To study the relationship between synovial membrane apoptosis and different arthropathy for the pathogenesis of cartilage lesion in arthropathy. METHODS: Synovial specimens (28 cases) were divided into control group (9 cases), osteoarthritis group (7 cases), dislocation group (5 cases), meniscus injury group (5 cases) and arthropyosis group (2 cases). Synovial histological sections were stained with methylene blue, and examined under microscope. DNA was extracted from synovial specimens, and analysed by gel electrophoresis. RESULTS: DNA ladder pattern was the most clear in the arthropyosis group, moderately clear in the dislocation group (especially in a case of 20 years-old shoulder dislocation), less clear in the osteoarthritis group, the least clear in the meniscus injury group, and was not seen in the control group. CONCLUSION: It can be concluded that the increased apoptosis is related to causes, course and seriously of arthropathy, and it might be a pathogenesis of cartilage lesion in arthropathy.

Major transmembrane movement associated with colicin Ia channel gating.

Colicin Ia, a bacterial protein toxin of 626 amino acid residues, forms voltage-dependent channels in planar lipid bilayer membranes. We have exploited the high affinity binding of streptavidin to biotin to map the topology of the channel-forming domain (roughly 175 residues of the COOH-terminal end) with respect to the membrane. That is, we have determined, for the channel's open and closed states, which parts of this domain are exposed to the aqueous solutions on either side of the membrane and which are inserted into the bilayer. This was done by biotinylating cysteine residues introduced by site-directed mutagenesis, and monitoring by electrophysiological methods the effect of streptavidin addition on channel behavior. We have identified a region of at least 68 residues that flips back and forth across the membrane in association with channel opening and closing. This identification was based on our observations that for mutants biotinylated in this region, streptavidin added to the cis (colicin-containing) compartment interfered with channel opening, and trans streptavidin interfered with channel closing. (If biotin was linked to the colicin by a disulfide bond, the effects of streptavidin on channel closing could be reversed by detaching the streptavidin-biotin complex from the colicin, using a water-soluble reducing agent. This showed that the cysteine sulfur, not just the biotin, is exposed to the trans solution). The upstream and downstream segments flanking the translocated region move into and out of the bilayer during channel opening and closing, forming two transmembrane segments. Surprisingly, if any of several residues near the upstream end of the translocated region is held on the cis side by streptavidin, the colicin still forms voltage-dependent channels, indicating that a part of the protein that normally is fully translocated across the membrane can become the upstream transmembrane segment. Evidently, the identity of the upstream transmembrane segment is not crucial to channel formation, and several open channel structures can exist.

Identification of a translocated protein segment in a voltage-dependent channel.

Voltage-gated channels undergo a conformational change in response to changes in transmembrane voltage. Here we use site-directed biotinylation to create conformation-sensitive sites on colicin Ia, a bacteriocidal protein that forms a voltage-sensitive membrane channel, which can be monitored by electrophysiological methods. We investigated a model of gating developed for the partly homologous colicin E1 that is based on the insertion of regions of the protein into the membrane in response to cis-positive voltages. Site-directed cysteine mutagenesis, followed by chemical modification, was used to attach a biotin molecule covalently to a series of unique sites on colicin Ia. The modified protein was incorporated into planar lipid membranes, where the introduced biotin moiety served as a site to bind the water-soluble protein streptavidin, added to one side of the membrane or the other. Our results show that colicin gating is associated with the translocation across the membrane of a segment of the protein of at least 31 amino acids.

Site-specific biotinylation of colicin Ia. A probe for protein conformation in the membrane.

Channel-forming colicins are Escherichia coli proteins that form voltage-dependent channels in lipid bilayer membranes and are lethal to sensitive strains of E. coli. Experiments with colicin E1 have led to a model of voltage dependence based on the insertion of alpha-helical segments of the protein into the membrane in response to cis-positive voltages. This model was tested on the partly homologous colicin Ia protein, which offers certain advantages over colicin E1 as a model channel, it is active at neutral pH and exhibits comparatively well-defined single channel conductance. We describe here the creation of a specific probe for locating a particular amino acid residue on one side or the other of a planar lipid bilayer membrane, by using the biotin-streptavidin system. Site-directed mutagenesis was used to change lysine 544 of colicin Ia to cysteine. This placed a unique cysteine at a site expected, by homology to colicin E1, to cross the membrane from the cis to the trans side in association with the opening of the channel. This unique cysteine was biotinylated chemically, so that it could serve as a target for streptavidin. Incubation of the biotinylated mutant colicin with streptavidin blocked its killing activity, in vivo; incubation of wild-type colicin, which lacks cysteine, with streptavidin, did not affect its activity. Channels formed by the biotinylated mutant protein in planar lipid bilayers were abolished by streptavidin added to the cis side of the membrane, if the channels were closed, but not if they were open. Trans streptavidin had no effect on either open or closed channels. Thus, when the channel is closed, residue 544 of colicin Ia is accessible to cis streptavidin in the closed state, but the opening of the channel eliminates this accessibility.