No abstract available.
Leprosy*

Enzymatic synthesis is an important way to produce β-alanine, but the biological method is expensive generally because of the high price of substrate. In this paper, a two-step enzymatic cascade system was developed, combining L-aspartase from Escherichia coli DH5α and L-aspartate α-decarboxylase from Corynebacterium glutamicum. This system catalyzes Fumarate and ammonia to β-alanine. The optimal ratio of AspA and PanD was 1:80 (W/W), and the concentration of AspA was 10 μg/mL, at 37 ℃ and pH 7.0. When the concentration of Fumarate was 100 mmol/L, the reaction reached its equilibrium after 8 h, and the yield of β-alanine was 90 mmol/L (7 g/L). The yield of β-alanine can reach 126 mmol/L (9.8 g/L) when the concentration of Fumarate increased to 200 mmol/L. Extending reaction time, the conversion rate did not change obviously. Using this two-step enzymatic cascade system, β-alanine from cheaper substrate Fumarate can be obtained.

Although there are 125 predicted DNase Ⅱ-like family genes in the Trichinella spiralis genome, plancitoxin-1-like (Ts-Pt) contains the HKD motif, a typical conserved region of DNase Ⅱ, in N- and C-terminal. It is generally believed that histidine is the active site in DNase Ⅱ. To study the nuclease activity of recombinant Ts-Pt with mutations in the active site from T. spiralis, different fragments of the mutated Ts-Pt genes were cloned using overlap PCR technique and inserted into the expressing vector pET-28a(+), and transformed into Escherichia coli Rosseta (DE3). The fusion proteins were purified by Ni-NTA affinity chromatography and SDS-PAGE. Nuclease activity of the recombinant proteins was detected by agarose gel electrophoresis and nuclease-zymography. The recombinant plasmids harboring the mutated Ts-Pt genes were constructed and expressed as inclusive body in a prokaryotic expression system. After renaturation in vitro, the recombinant proteins had no nuclease activity according to agarose gel electrophoresis. However, the expressed proteins as inclusive body displayed the ability to degrade DNA after renaturation in gel. And the nuclease activity was not affected after subjected to mutation of active site in N- and C-termini of Ts-Pt. These results provide the basis to study the relationship between DNase Ⅱ-like protein family and infection of T. spiralis.

OBJECTIVETo compare the effects of flipping moxibustion ofmedicine combined with western medication and simple western medication for rheumatoid arthritis with cold dampnesssyndrome.

METHODSEighty patients were randomly assigned into an observation group and a control group, 40 cases in each group. Oral methotrexate (1 time a week) and sulfasalazine (twice a day except the day for methotrexate) were used in the two groups. Patients in the observation group were treated with flipping moxibustion ofmedicine, twice a week. The main acupoints were Shenshu (BL 23), Guanyuan (CV 4), Zusanli (ST 36), Yinlingquan (SP 9), and the matching acupoints were in the meridians related to the disease location. All the treatment was given for continuous 4 weeks. The TCM symptom score, visual analogue scale (VAS) score, blood sedimentation (ESR), rheumatoid factor (RF) and C-reactive protein (CRP) were observed before and after treatment. The effect was evaluated.

RESULTSThe total effect rate in the observation group was 95.0% (38/40), which was better than 77.5% (31/40) in the control group (<0.05). After treatment, the VAS score, TCM symptom score, RF, ESR, CRP levels decreased in the two groups (all<0.05), with better effects in the observation group (all<0.05). The adverse reactions in the observation group were lower than those in the control group.

CONCLUSIONFlipping moxibustion ofmedicine combined with western medication for rheumatoid arthritis with cold dampnesssyndrome are better than simple western medication, which are safer and more effective.

Euphorbia factor L2, a lathyrane diterpenoid isolated from caper euphorbia seed (the seeds ofL.), has been traditionally applied to treat cancer. This article focuses on the cytotoxic activity of Euphorbia factor L2 against lung carcinoma A549 cells and the mechanism by which apoptosis is induced. We analyzed the cytotoxicity and related mechanism of Euphorbia factor L2 with an MTT assay, an annexin V-FITC/PI test, a colorimetric assay, and immunoblotting. Euphorbia factor L2 showed potent cytotoxicity to A549 cells. Euphorbia factor L2 led to an increase in reactive oxygen species (ROS) generation, a loss of mitochondrial electrochemical potential, release of cytochromeactivation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase, suggesting that Euphorbia factor L2 induced apoptosis through a mitochondrial pathway. The cytotoxic activity of Euphorbia factor L2 in A549 cells and the related mechanisms of apoptotic induction provide support for the further investigation of caper euphorbia seeds.

OBJECTIVETo explore the impact of neutrophil gelatinase-associated lipocalin (NGAL) knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC) on the proliferation, migration and apoptosis of human colon cancer cells.

METHODSNGAL siRNA was encapsulated by UAC and chitosan (CTS) respectively, and then was transfected into human colon cancer cell lines HT29. The NGAL mRNA was detected by real-time quantitative PCR (RT-QPCR). Relationships of NGAL gene silencing with the proliferation, migration and apoptosis of HT29 cell were analyzed.

RESULTSUnder the fluorescence microscope, the transfection efficiency of siRNA in UAC group was (37.52±7.17)%, which was significantly higher than (11.32±3.39)% in CTS group (t=6.102, P=0.005). Forty-eight hours after transfection, RT-QPCR examination showed that the level of NGAL mRNA expression was 0.350 in UAC group and 0.529 in CTS group with significant difference (t=-3.743, P=0.02), meanwhile both levels were significantly lower as compared to control group(F=163.538, P<0.001). Proliferation analysis revealed that after silencing NGAL gene, proliferation rate of UAC group and CTS group was slightly lower than control group, and no significant differences were found (F=9.520, P=0.438). However, migration assay demonstrated that the 24-hour migration rate of UAC group and CTS group was significantly lower than that of control group (F=6.756, P=0.029), meanwhile the migration rate of UAC group was slightly lower than that of CTS group [(77.90±7.14)% vs. (87.67±3.98)%, t=-1.704, P=0.164]. Apoptosis detection revealed that the apoptosis rate in UAC group was significantly higher than that in CTS group and the control group 2 days after transfection [(15.800±1.054)% vs. (12.900±0.656)%, (11.933±1.914)%, F=7.004, P=0.027].

CONCLUSIONSThe encapsulated ability and transfection efficiency of chitosan modified by urocanic acid elevate significantly. Silencing NGAL gene by UAC carrier can down-regulate the expression of NGAL mRNA in HT29 colon cell line, inhibit their migration and facilitate their apoptosis.

Objective To summarize our experiences in the diagnosis and prognosis of different subtypes of primary thyroid lymphoma (PTL). Methods The clinical data of 27 PTL patients who were treated in our hospital from January 1998 to December 2014 were retrospectively analyzed. The pathological types of these patients included B cell lymphoma unclassifiable (BCLU) (n=5),mucosa-associated lymphiod tissue lymphoma (MALT) (n=9),diffuse large B cell lymphoma (DLBCL) (n=12),and T cell lymphoma (n=1). Results Of all these 27 cases,the most common clinical symptom was painless swelling of the neck (n=21,77.8%). Of 7 patients who had received preoperative fine needle biopsy,lymphoma was suspected in 2 cases (28.6%). Among these 7 cases,the positive rate of suspicious lymphoma was 66.7% in 3 DLBCL patients,0 in 3 MALT patients,and 0 in 1 BCLU paitent. Also,25 patients underwent intraoperative frozen pathological examination,which revealed lymphoma or suspicious lymphoma in 16 cases (64.0%); in these patients,the positive rate was 66.7% for BCLU,77.8% for MALT,58.3% for DLBCL,and 0 for T-cell lymphomas. The overall survival was (89.3±12.4) months,and the overall 5-year survival rate was 61.6%. The estimated survival in symptomatic group was 31.6 months,which was significantly shorter than that in asymptomatic group (97.9 months) (P=0.032). Other factors including age,sex,tumor size,tumor stage,international prognostic index,tracheal compression,lactate dehydrogenase,residual tumor,and pathological type showed no significant effect on survival(all P>0.05). Conclusions DLBCL has the highest fine needle biopsy positive rate,MALT has the highest frozen pathological positive rate,and intraoperative frozen pathology has more malignant results than the preoperative fine needle biopsy in the diagnosis. The accompanying lymphoma symptoms may be an adverse prognostic factor.

As a kind of intervention measures of traditional Chinese medicine, acupuncture-moxibustion is highly adopted on global clinical practice. Even though the global clinical trial registration system was established more than 10 years ago, the proportion of acupuncture-moxibustion clinical trial registration is still very low; and it is very problematic on the methodological quality and report quality in the published acupuncture-moxibustion clinical trials. In order to manage particularly the acupuncture-moxibustion clinical trials, China Academy of Chinese Medical Sciences, collaborated with China Association of Acupuncture and Moxibustion and World Federation of Acupuncture Societies, established the Acupuncture-Moxibustion Clinical Trail Registry (AMCTR). AMCTR is a secondary registry platform affiliated to the Chinese Clinical Trial Registry (ChiCTR) and WHO International Clinical Trials Registry Platform (ICTRP), specifically for the acceptance and management of clinical trials in the field of acupuncture and moxibustion. It is a nonprofit academic organization, located in China Academy of Chinese Medical Sciences.

OBJECTIVETo correlate the clinical features of patients with acute myeloid leukemia (AML) with mutations of FLT3-ITD, NPM1, CEBPA, c-KIT, DNMT3A and ND4 genes as well as chromosomal aberrations.

METHODSSomatic mutations of aforementioned genes in 412 newly diagnosed AML patients were detected with PCR and direct sequencing. All patients were also subjected to R-banding chromosomal analysis. The results were correlated with the clinical features and prognosis of the patients.

RESULTSThe mutation rates of FLT3-ITD, NPM1, CEBPA, c-KIT, DNMT3A and ND4 were 9.0% (26/289), 19.1% (50/262), 18.9% (34/180), 3.4% (7/208), 6.6% (9/137) and 6.9% (4/58), respectively. Patients with poor prognosis based on genetic mutations had lower blood platelet count than those with intermediate and good prognosis (P=0.001 and P=0.001, respectively). None of the three groups attained median overall survival (OS) (P> 0.05). The complete remission (CR) was similar among the three groups (P> 0.05). For patients with different prognosis based on cytogenetic findings, white blood cell count in those with intermediate prognosis was higher than those with good and poor prognosis (P< 0.001 and P=0.004, respectively), while the blood platelet count of the intermediate group was higher than that of the group with good prognosis (P=0.018). No significant difference was found among the three groups in terms of hemoglobin level (P> 0.05). The group with poor prognosis has attained shorter OS compared with those with good and intermediate prognosis (P< 0.001 and P=0.003, respectively). However, the CR rate of the group with good prognosis was higher than that of the intermediate group (P=0.001). For the group with intermediate prognosis, presence of genetic mutations did not correlate with the clinic characteristics such as white blood cell count, blood platelet count, hemoglobin level, OS and CR rate (P> 0.05 for all comparisons).

CONCLUSIONGenetic mutations combined with cytogenetic analysis can facilitate the prognosis and personalized treatment for patients with AML.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukemia, Myeloid, Acute ; genetics ; mortality ; Male ; Middle Aged ; Mutation ; Prognosis ; Young Adult

Members of the genus Anaplasma are important emerging tick-borne pathogens in both humans and animals in tropical and subtropical areas. Here, we investigated the presence of Anaplasma spp. in 621 sheep and 710 goats from six provinces of China. Polymerase chain reaction (PCR) and DNA sequencing were conducted to determine the prevalence of Anaplasma (A.) phagocytophilum, A. ovis and A. bovis targeting the 16S ribosomal RNA or the major surface protein 4 gene. PCR revealed Anaplasma in 39.0% (240/621) of sheep and 45.5% (323/710) of goats. The most frequently detected species was A. ovis (88/621, 14.2% for sheep; 129/710, 18.2% for goats), followed by A. bovis (60/621, 9.7% for sheep; 74/710, 10.4% for goats) and A. phagocytophilum (33/621, 5.3% for sheep; 15/710, 2.1% for goats). Additionally, eight sheep and 20 goats were found to be infected with three pathogens simultaneously. DNA sequencing confirmed the presence of these three Anaplasma species in the investigated areas, and phylogenetic analysis indicated that there was geographic segregation to a certain extent, as well as a relationship between the host and cluster of A. ovis. The results of the present study provide valuable data that helps understand the epidemiology of anaplasmosis in ruminants from China.
Anaplasma ovis ; Anaplasma phagocytophilum ; Anaplasma* ; Anaplasmosis ; Animals ; China* ; Epidemiology ; Goats* ; Humans ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S ; Ruminants ; Sequence Analysis, DNA ; Sheep*