ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy， to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian， and meanwhile， the contents of four index components（geniposide， paeoniflorin， hesperidin and paeonol） were quantitatively analyzed by high performance liquid chromatography（HPLC）. Nonalcoholic fatty liver disease（NAFLD） rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group， model group， lovastatin group（1.8 mg·kg^-1）， combined decoction group（1.26 g·kg^-1） and single decoction group（1.18 g·kg^-1）. After successful modeling， lovastatin group， combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day， and the control group and model group were given equal amounts of normal saline by intragastric administration， after 4 weeks of administration， the serum and liver tissues were collected， and the contents of alanine aminotransferase（ALT）， aspartate aminotransferase（AST）， total cholesterol（TC）， triglyceride（TG）， low-density lipoprotein cholesterol（LDL-C） and high-density lipoprotein cholesterol（HDL-C） in serum of rats were detected， and the liver pathological examination was carried out by hematoxylin-eosin（HE） staining and oil red O staining， so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian， and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide， paeoniflorin， hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples（P<0.01）. The pharmacodynamic results showed that compared with the model group， both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats， reduce the serum levels of AST， ALT， TC， TG and LDL-C， increase the serum level of HDL-C， and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However， there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition， but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.

Objective To investigate the relationship between left ventricular hypertrophy(LVH)diagnosed by Peguero-Lo-Presti index and recurrence of paroxysmal atrial fibrillation(AF)after radiofrequency ablation.Methods A total of 652 patients with paroxysmal atrial fibrillation who underwent radiofrequency ablation were selected.According to Peguero-Lo-Presti index,patients were divided into the LVH group(167 cases)and the normal left ventricle group(485 cases).Baseline data were collected,and regular follow-up was performed at 3,6 and 12 months after radiofrequency catheter ablation.The recurrence of AF was assessed.Kaplan-Meier survival curve was used to analyze the recurrence rate of AF in the two groups.Cox proportional hazard model was used to assess risk factors for recurrent atrial fibrillation.Results The median follow-up time was 20.5(15.0,26.0)months.A total of 155 patients(23.8%)developed recurrence of AF,including 95 patients in the LVH group and 60 patients in the LVN group.The recurrence rate without AF was significantly lower in the LVH group than that in the LVN group(64.1%vs.80.4%,Log-rank χ2=26.361,P＜0.01).After adjusting for age,sex,body mass index,hypertension,diabetes,coronary heart disease,cardiac dysfunction,left anteroposterior and posterior atrial diameter,left ventricular end-diastolic diameter,and left ventricular ejection fraction,LVH diagnosed by Peguero-Lo-Presti index was still a risk factor for recurrent AF[HR(95%CI):2.359(1.663-3.345),P＜0.01].Conclusion In patients with paroxysmal AF,LVH diagnosed by Peguero-Lo-Presti index is a risk factor of AF recurrence after radiofrequency catheter ablation.

Objective:A case-control association analysis was performed to investigate if the single nucleotide polymorphisms (SNPs) of N-cadherin(CDH2) gene is implicated in schizophrenia in a Han Chinese population.Methods:A total of 528 patients with paranoid schizophrenia and 528 healthy controls were recruited from northern Henan province to analyze 25 SNPs located in CDH2 gene.The clinical symptoms of 267 first-episode schizophrenia patients were evaluated with positive and negative syndrome scale (PANSS), and the correlation between CDH2 gene and clinical symptoms was analyzed by SNPStats software online.Results:Allele frequencies of rs9951577 and rs1231268 were significantly correlated with schizophrenia( P<0.05), genotype frequency of rs1639387 was significantly correlated with schizophrenia( P=0.044). After gender classification, SNPs rs1789470 and rs28365328 were found to be significantly correlated with schizophrenia in female patients ( P=0.044, 0.019). In addition, the study found that CDH2 was correlated with the clinical characteristics of schizophrenia( P<0.05), and the negative factor score of patients between GG type rs1231268 and the other two genotypes (AG+ AA) ((21.12±8.41) vs (18.87±7.52)) was statistically significant ( P<0.05). Conclusion:CDH2 gene may be one of the susceptibility genes to SZ, and has definite correlation with clinical negative symptoms.

Leucine dehydrogenase (LDH) is the key rate-limiting enzyme in the production of L-2-aminobutyric acid (L-2-ABA). In this study, we modified the C-terminal Loop region of this enzyme to improve the specific enzyme activity and stability for efficient synthesis of L-2-ABA. Using molecular dynamics simulation of LDH, we analyzed the change of root mean square fluctuation (RMSF), rationally designed the Loop region with greatly fluctuated RMSF, and obtained a mutant EsLDHD2 with a specific enzyme activity 23.2% higher than that of the wild type. Since the rate of the threonine deaminase-catalyzed reaction converting L-threonine into 2-ketobutyrate was so fast, the multi-enzyme cascade catalysis system became unbalanced. Therefore, the LDH and the formate dehydrogenase were double copied in a new construct E. coli BL21/pACYCDuet-RM. Compared with E. coli BL21/pACYCDuet-RO, the molar conversion rate of L-2-ABA increased by 74.6%. The whole cell biotransformation conditions were optimized and the optimal pH, temperature and substrate concentration were 7.5, 35 °C and 80 g/L, respectively. Under these conditions, the molar conversion rate was higher than 99%. Finally, 80 g and 40 g L-threonine were consecutively fed into a 1 L reaction mixture under the optimal conversion conditions, producing 97.9 g L-2-ABA. Thus, this strategy provides a green and efficient synthesis of L-2-ABA, and has great industrial application potential.
Aminobutyrates ; Escherichia coli/genetics* ; Leucine Dehydrogenase/genetics* ; Threonine Dehydratase

5-aminolevulinic acid (5-ALA) plays an important role in the fields of medicine and agriculture. 5-ALA can be produced by engineered Escherichia coli and Corynebacterium glutamicum. We systematically engineered the C4 metabolic pathway of C. glutamicum to further improve its ability to produce 5-ALA. Firstly, the hemA gene encoding 5-ALA synthase (ALAS) from Rhodobacter capsulatus and Rhodopseudomonas palustris were heterologously expressed in C. glutamicum, respectively. The RphemA gene of R. palustris which showed relatively high enzyme activity was selected. Screening of the optimal ribosome binding site sequence RBS5 significantly increased the activity of RphemA. The ALAS activity of the recombinant strain reached (221.87±3.10) U/mg and 5-ALA production increased by 14.3%. Subsequently, knocking out genes encoding α-ketoglutarate dehydrogenase inhibitor protein (odhI) and succinate dehydrogenase (sdhA) increased the flux of succinyl CoA towards the production of 5-ALA. Moreover, inhibiting the expression of hemB by means of sRNA reduced the degradation of 5-ALA, while overexpressing the cysteine/O-acetylserine transporter eamA increased the output efficiency of intracellular 5-ALA. Shake flask fermentation using the engineered strain C. glutamicum 13032/∆odhI/∆sdhA-sRNAhemB- RBS5RphemA-eamA resulted in a yield of 11.90 g/L, which was 57% higher than that of the original strain. Fed-batch fermentation using the engineered strain in a 5 L fermenter produced 25.05 g/L of 5-ALA within 48 h, which is the highest reported-to-date yield of 5-ALA from glucose.
Aminolevulinic Acid/metabolism* ; Corynebacterium glutamicum/metabolism* ; Fermentation ; Metabolic Engineering ; Rhodobacter capsulatus/enzymology* ; Rhodopseudomonas/enzymology*

Objective To analyze the residents' iodine nutrition level of Yunfu City in Guangdong Province from 2009 to 2016,in order to provide evidence for making prevention strategy of iodine deficiency disorders.Methods Four surveys on salt iodine levels,four surveys on urinary iodine levels of children aged 8-10 years old,two surveys on urinary iodine levels of pregnant women and one survey on iodine content in drinking water were carried out according to the prevention and control planning for endemic disease in Guangdong Province in Yunfu City from 2009 to 2016.Salt iodine was determined by direct titration (GB/T 13025.7-1999,GB/T 13025.7-2012),urinary iodine was determined by arsenic cerium catalytic spectrophotometry (WS/T 107-2006,WS/T 107.1-2016),water iodine was determined by the national iodine deficiency disease reference laboratory recommended method.Results In 2009,2011,2015 and 2016,1 464,1 464,1 500,1 500 salt samples and 500,388,1 000,1 000 children were investigated,the medians of salt iodine levels were 30.3,28.1,24.8,and 24.8 mg/kg,respectively,and the medians of children urinary iodine levels were 208.3,188.3,143.1 and 165.2 μg/L,respectively.Five hundred,five hundred pregnant women were investigated in 2015 and 2016,and the medians of pregnant women urinary iodine levels were 101.0 and 96.4 μg/L,respectively.A total of 1 149 samples of drinking water were investigated in 2011-2016,the median of water iodine was 2.8 μg/L.Conclusion Yunfu is an iodine deficiency city,after adjusting the iodine content of edible salt in 2012,iodine nutrition in children is at an appropriate level,but the iodine intake level of pregnant women is relatively lower,so prevention and treatment of iodine deficiency disorders and urinary iodine monitoring should be strengthened in pregnant women.

Objective To evaluate the potential cerebral cortical volume alterations in type 2 diabetes mellitus (T2DM) patients with mild cognitive impairment (MCI) compared with T2DM patients and healthy controls,and to observe the correlations with the scores of neuropsychological scales.Methods Cortical volume based on high-resolution MR T1WI data from 30 healthy controls (HC),30 T2DM patients and 30 T2DM with MCI patients were evaluated with FreeSurfer software and compared with variance analysis.The correlations between cerebral cortical volume which had statistical difference and the scores of neuropsychological scales were analyzed.Results There were significant differences in auditory verbal learning test (AVLT) scores,complex figure test-delayed recall (20 min) scores,digit symbol-coding subtest scores,MoCA scores and higher trail-making test-A scores,as well as trail-making test-B scores between T2DM and T2DM with MCI patients (all P＜0.05).Compared with T2DM patients,cortical volume of left entorhinal cortex,left lateral orbitofrontal gyrus,left posterior cingulate gyrus and the right lateral orbitofrontal gyrus,right pars orbitalis,right insula reduced in T2DM with MCI patients (all P＜0.05).In T2DM with MCI patients,AVLT scores were positively correlated with volume of the left entorhinal cortex (r=0.452,P=0.018).Conclusion Several cortical volume reductions are exhibited in T2DM patients with MCI.The volume of the left entorhinal cortex may be a potential biomarker to diagnose and evaluate MCI in T2DM.

OBJECTIVETo investigate the effect and safety of acupuncture with regulating menstruation to promote pregnancy for diminished ovarian reverse (DOR).

METHODSAccording to prospective case series, 46 patients were observed and finally 40 cases were included. The acupoints were ① Baihui (GV 20), Shenting (GV 24), Guanyuan (CV 4) and bilateral Benshen (GB 13), Huangshu (KI 16), Dahe (KI 12), Luanchao (Extra), Zusanli (ST 36), Sanyinjiao (SP 6), Taixi (KI 3), Taichong (LR 3) and ② bilateral Shenshu (BL 23) and Ciliao (BL 32). The points in the two groups were used alternately. Acupuncture was given for 3 courses, 12 times as a course and 3 times a week. Before and after treatment, and 3 months after treatment, follicle-stimulating hormone (FSH), follicle-stimulating hormone/luteinizing hormone (FSH/LH), estradiol (E), antral follicle count (AFC) and TCM symptom score were observed. The safety was evaluated.

RESULTSCompared with before treatment, the levels of FSH, FSH/LH decreased, and the levels of Eand AFC increased after treatment and at follow-up (all<0.05). And the TCM symptom scores were significantly lower than those before treatment (both<0.05). The rate of pregnancy after treatment was 15% (6/40). There was no infection and organ injury.

CONCLUSIONAcupuncture with regulating menstruation to promote pregnancy can safely improve the ovarian reserve of patients with DOR.

BACKGROUND:Drug therapy can partly reduce and delay the progress of Alzheimer’s disease, but only 30%with the single drug treatment obtain clinical cure. OBJECTIVE:To study the therapeutic effect of bone marrow mesenchymal stem cel s transplantation for rats with Alzheimer’s disease. METHODS:Amyloidβ-protein was injected into the hippocampus of Sprague-Dawley rats to construct the model of Alzheimer’s disease. And bone marrow stromal stem cel s were transplanted into the hippocampus of the rat models. RESULTS AND CONCLUSION:At 2 weeks after modeling, compared with the control group, the escape latency in the model and experimental groups was significantly longer (P<0.05), which indicating that Alzheimer’s disease models were successful y established. At 4 weeks after cel transplantation, compared with the model group, the average escape latency in the experimental group was significantly decreased, but retention time on the platform quadrant was significantly prolonged (P<0.05). Besides, at 4 weeks after cel transplantation, expression of choline acetyltransferase in the experimental group was significantly higher than that in the model group (P<0.05). In conclusion, bone marrow mesenchymal stem cel s cannot only differentiate and survive in the hippocampus of rats with Alzheimer’s disease, but also improve the learning and memory ability.

BACKGROUND:Drug treatment for senile dementia has unsatisfactory outcomes although to a certain extent it can reduce and delay the progression of Alzheimer’s disease. Stem cel transplantation is a new attempt for the treatment of senile dementia.
OBJECTIVE:To observe the effect of bone marrow mesenchymal stem cel transplantation on the behavior of senile dementia rats.
METHODS: Rat models of senile dementia were made in 20 Sprague-Dawley rats that were given continuous 60-day gavage of aluminium chloride solution. Then, model rats were randomized into model group treated with normal saline injection and experimental group treated with hippocampal injection of bone marrow mesenchymal stem cels, respectively. Another 10 rats undergoing normal feeding served as control group. Learning and memory ability of rats were tested by Morris water maze, and superoxide dismutase activity and malondialdehyde content in brain tissues of rats were measured by colorimetric method at 4 weeks after cel transplantation.
RESULTS AND CONCLUSION:Compared with the model group, the escape latency was shortened and the cross-platform frequency was increased in the experimental group (P < 0.05), and moreover, significantly elevated superoxide dismutase activity and reduced malondialdehyde content in the brain tissues of rats were found in the experimental group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cel transplantation contributes to behavior improvement in senile dementia rats by improving the learning and memory ability.