Sialyllactose is one of the most abundant sialylated oligosaccharides in human milk oligosaccharides (HMOs), which plays an important role in the healthy development of infants and young children. However, its efficient and cheap production technology is still lacking presently. This study developed a two-step process employing multiple-strains for the production of sialyllactose. In the first step, two engineered strains, E. coli JM109(DE3)/ pET28a-BT0453 and JM109(DE3)/pET28a-nanA, were constructed to synthesize the intermediate N-acetylneuraminic acid. When the ratio of the biomass of the two engineered strains was 1:1 and the reaction time was 32 hours, the maximum yield of N-acetylneuraminic acid was 20.4 g/L. In the second step, E. coli JM109(DE3)/ pET28a-neuA, JM109(DE3)/ pET28a-nst and Baker's yeast were added to the above fermentation broth to synthesize 3'-sialyllactose (3'-SL). Using optimal conditions including 200 mmol/L N-acetyl-glucosamine and lactose, 150 g/L Baker's yeast, 20 mmol/L Mg²⁺, the maximum yield of 3'-SL in the fermentation broth reached 55.04 g/L after 24 hours of fermentation and the conversion rate of the substrate N-acetyl-glucosamine was 43.47%. This research provides an alternative technical route for economical production of 3'-SL.
Child ; Humans ; Child, Preschool ; N-Acetylneuraminic Acid ; Escherichia coli/genetics* ; Lactose ; Fermentation ; Saccharomyces cerevisiae ; Oligosaccharides ; Glucosamine

Objective To explore the diagnostic value of Young's modulus obtained by real-time shear wave elastography (SWE) for liver fibrosis in autoimmune hepatitis (AIH) patients. Methods A total of 75 AIH patients in the First Affiliated Hospital of Zhengzhou University from January 2013 to April 2022 were retrospectively enrolled. Scheuer scoring system was used to evaluate degrees of liver fibrosis (S0-S4). By using pathological examination of liver tissues as the golden standard, the receiver operating characteristic curve (ROC) was plotted and the area under the curve (AUC) was used to evaluate the diagnostic value of SWE for the significant fibrosis (≥S2), advanced liver fibrosis (≥S3), and liver cirrhosis (S4), respectively. Independent sample t test was used for comparison of continuous data with normal distribution between the two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and Bonferroni method was used for further comparison between two groups. The Spearman correlation coefficient was used for correlation analysis. The logistic regression analysis was used to predict the impact factors in diagnosis accuracy. Results The Young's modulus measured by SWE was statistically significant different among various fibrosis groups ( H =35.186, P < 0.001) although there was no statistical significance in patients' age and platelet, alanine aminotransferase, aspartate aminotransferase, total bilirubin, alkaline phosphatase, and glutamyl transpeptidase levels (all P > 0.05). The Young's modulus measurement was positively correlated with liver fibrosis ( r =0.675, P < 0.05). The AUCs of SWE in the diagnosis of≥S2, ≥S3, and S4 were 0.839, 0.820 and 0.898, respectively and the corresponding optimum cut-off values were 9.2, 10.9, and 14.4 kPa, respectively. The overall concordance rate of the liver Young' s modulus measurements vs . fibrosis stages was 57.33%. Moreover, the alkaline phosphatase level was an independent predictor for diagnostic accuracy of SWE for stage 0-1 fibrosis ( OR =1.009, 95% CI : 1.001-1.018, P =0.029). Conclusion The SWE possessed a diagnosis value for the significant fibrosis (≥S2), advanced liver fibrosis (≥S3) and liver cirrhosis (S4), although there was a low overall concordance rate in the liver Young's modulus measurements obtained using SWE vs. fibrosis stages.

Ankylosing spondylitis (AS) is a chronic inflammatory disease mainly affecting the sacroiliac joints, spine and peripheral joints. In China, standardized diagnosis and treatment of AS is still to be popularized. Based on the evidence and guidelines from China and other countries, Chinese Rheumatology Association developed standardization of diagnosis and treatment of AS. The purposes are: (1) to standardize the diagnosis and evaluation of AS; (2) to promote rational use of non-steroidal anti-inflammatory drugs, biological as well as traditional disease modifying anti-rheumatic drugs, so as to improve the patient′s quality of life.

Objective To analyze the pathogenic characteristics and resistance of severe lung infection patients in Suining area, and to provide a reference for early clinical intervention. Methods A total of 359 patients with pulmonary infection were selected in Suining city from December 2019 to December 2020.The patients were divided into mild group (231 cases) and severe group (128 cases) according to the severity of nosocomial pneumonia.Sputum samples were collected for pathogen identification and drug sensitivity test. Multivariate logistic regression was used to analyze the influencing factors of severe pulmonary infection. Results There were 128 cases of severe pneumonia in 359 patients with traumatic brain injury. 147 strains of pathogenic bacteria were isolated, including 91 strains (61.90%) of gram-negative bacteria, mainly including 42 strains (28.57%) of Acinetobacter baumannii, 29 strains (19.73%) of Pseudomonas aeruginosa and 16 strains (10.88%) of Klebsiella pneumoniae. There were 37 gram-positive strains (25.17%), and 31 strains (21.09%) were staphylococcus aureus. A total of 19 strains of fungi (12.93%); Acinetobacter baumannii and Klebsiella pneumoniae were more sensitive to cefoperazone sodium sulbactam sodium and iaropenem, and more resistant to benzylbenzicillin, sulfamethoxazole/trimethoprazine and cefazolin. Pseudomonas aeruginosa had higher sensitivity to ceftazidine, piperacillin/tazobactam, and higher resistance to cefazolin sodium, ampicillin and other drugs. Staphylococcus aureus was highly sensitive to linezolid, vancomycin and teicoranin, and had high resistance to cefazolin and penicillin. There were statistically significant differences between the two groups in age, GCS score, combined basic diseases >2, coma time >24h, hypoproteinemia and invasive operation (P<0.05). Logistic regression analysis showed that GCS score, hypoproteinemia and invasive operation were independent risk factors for severe pulmonary infection in patients with traumatic brain injuries (P<0.05). Conclusion Suining patients with gram-negative bacteria, mainly Acinetobacter baumannii, Pseudomonas aeruginosa, aeruginosa, should be based on the pathogen susceptibility results of patients, for invasive operation, high GCS score should be positive treatment, and actively correct hypoproteinemia, can reduce the occurrence of severe lung infection.

Pelvic radiotherapy is a way for treatment of most pelvic tumors, of which the pelvic insufficiency fracture (PIF) is a long-term complication. In this review, research progress of pelvic insufficiency fracture is summarized and discussed. The pathogenesis of PIF is mainly about inhibition of osteoblasts and the risk factors of PIF include old age, postmenopausal status, absence of hormonal replacement therapy, high number of births, smoking history, low body mass index (BMI), concurrent rheumatoid arthritis, concurrent diabetes mellitus, intracavitary brachytherapy of the high dose rate (HDR-ICBT), high dose of radiotherapy, etc. Effective drugs for prevention or treatment of PIF have not been found yet. Delayed diagnosis and misdiagnosis of PIF can cause serious consequences. As a result, further studies are needed to guide clinical work.

【Objective】 To explore the effect of Genistein on the proliferation and cell cycle regulation of ovarian cancer cells. 【Methods】 Ovarian cancer OVCAR-5 cells were treated with Genistein. Cell counting and MTS assays were performed to determine the alterations of cell proliferation. Real-time PCR and Western blotting were conducted to examine the expression changes of key cell cycle regulators. 【Results】 Genistein significantly promoted the proliferation and viability of OVCAR-5 cells. After Genistein treatment, cellular mRNA and protein expression levels of cell cycle activators such as PCNA, Cyclin D1 and CDK4 were increased, but those of cell cycle inhibitors such as p21 and p27 were decreased. 【Conclusion】 Genistein can upregulate the proliferation and G1-S transition of ovarian cancer OVCAR-5 cells. The discrepancy may be caused by diverged experimental conditions and/or different ER expression patterns of cell lines. The findings may provide basic information for in-depth analysis of the role(s) and mechanisms by which genistein confers its effect on ovarian cancer cells.

Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.

Excess activation of cardiac fibroblasts inevitably induces cardiac fibrosis. Emodin has been used as a natural medicine against several chronic diseases. The objective of this study is to determine the effects of emodin on cardiac fibrosis and the underlying molecular mechanisms. Intragastric administration of emodin markedly decreased left ventricular wall thickness in a mouse model of pathological cardiac hypertrophy with excess fibrosis induced by transaortic constriction (TAC) and suppressed activation of cardiac fibroblasts induced by angiotensin II (AngII). Emodin upregulated expression of metastasis associated protein 3 (MTA3) and restored the MTA3 expression in the setting of cardiac fibrosis. Moreover, overexpression of MTA3 promoted cardiac fibrosis; in contrast, silence of MTA3 abrogated the inhibitory effect of emodin on fibroblast activation. Our findings unraveled the potential of emodin to alleviate cardiac fibrosis upregulating MTA3 and highlight the regulatory role of MTA3 in the development of cardiac fibrosis.

Objective To investigate the effects of mild hypothermia on post-resuscitation neurological outcome after ventricular fibrillation (VF) in rabbits.Methods Forty-five adult New Zealand rabbits were induced VF by direct current of electricity.The rabbits were randomly(random number) divided into following groups:normothermic resuscitation group (NR),mild hypothermia prearrest group (HP),mild hypothermia resuscitation 30 min group (HRe30),mild hypothermia resuscitation 90 min group (HRe90),normothermic sham group (NS),and hypothermia sham group (HS).The rabbits of NR group were observed for 600 min in room temperature after restoration of spontaneous circulation (ROSC).The mild hypothermia was induced by surface cooling,and maintained for 600 min after the aimed low temperature reached.The arterial blood samples were collected for determining neuron-specific enolase (NSE) and thioredoxin (Trx) and the mean arterial pressure (MAP),left ventricular end-diastolic pressure (LVEDP) and left ventricular pressure raise and fall rate (±dp/dtmax) were observed at 15 min before CA,and 30 min,60 min,120 min,360 min and 600 min after ROSC.After the animals were sacrificed at 600 min after ROSC,the whole brain of animals was harvested and observed under light microscope to calculate the apoptotic index of the hippocampal CA1 neurons by using TUNEL method.One-way ANOVA was used to determine the statistical significance between two groups,a two-tailed value of P＜0.05 was considered statistically significant.Results (1) Hemodynamically compared with normal temperature groups,HR was lower in hypothermia groups.Compared with NR,HRe30,and HRe90 group,LVEDP was higher in HP group at 30 min after ROSC(3.4±0.8 vs.4.6±1.0,4.1±0.5,4.3±0.2,F=9.85,P=0.019).In Hp group,the level of +dp/dtmax was higher than that in NR,HRe30 and HRe90 groups at 30 min and 120 min after ROSC.In HP group,the level of-dp/dtmax was higher than that of NR group at 30 min,60 min,120 min,360 min and 600 min after ROSC.(2) Serologically compared with HP,HRe30 and HRe90 group,NSE levels were higher in NR group at 60 min,120 min and 360 min after ROSC.Compared with NR,HRe30,and HRe90 group,Trx levels in NR group were lower at 60 min,120 min,360 min and 600 min after ROSC.Compared with HP group,Trx levels in HRe30 and HRe90 groups were higher at 60 min,120 min,360 min and 600 min after ROSC.(3) Pathologically compared with NR group,histopathological changes in hippocampus CA1 area were milder found in HP,HRe30 and HRe90 groups.AI (％) was lower in HP,HRe30 and HRe90 groups than that in NR group[(62.25±10.43)％ vs.(20.61±5.02)％,(25.08±3.92)％,(30.33±7.15)％,P=0.001].Concusions This study shows that hypothermia should be initiated as soon as possible,and especially early intra-arrest cooling appears to be significantly better than post-ROSC cooling and normothermia.

Objective To analyze the relationship between the posterior circulation cerebral infarction and posterior embryonic cerebral artery,and to explore the possibility of posterior embryonic cerebral artery to be predictive indicator for the occurrence and development of the posterior circulation infarction.Methods A total of 2341 posterior circulation cerebral infarction patients in our hospital were recruited,and its location was confirmed in infarction circulation blood supply range and existence condition of posterior embryonic cerebral artery was explored by head MRI and DWI.The correlation between posterior embryonic cerebral artery and posterior circulation cerebral infarction,arteriosclerosis.Results Among the 2341 patients,there were 1012(41.63％) patients with posterior circulation infarction,including 314 cases with left posterior circulation infarction,295 cases with right posterior circulation infarction,403 cases with bilateral posterior circulation infarction.There were 578 (23.78％) patients with embryonic posterior cerebral artery,including 179 cases with left posterior cerebral artery,257 cases with right posterior cerebral artery,and 142 cases with bilateral posterior cerebral artery.There were 1193 (49.1％) patients with cerebral arteriosclerosis.The incidences of cerebral arteriosclerosis and posterior circulation cerebral infarction showed significant difference in the embryo posterior cerebral artery and non-embryo posterior cerebral artery patients (P ＜ 0.05),but no significant difference in posterior embryonic cerebral artery patients with different locations (P ＞ 0.05).Conclusion Embryonic posterior cerebral artery has a significant correlation with posterior circulation cerebral infarction,and can significantly increase the incidence of posterior circulation cerebral infarction.