Total RNA was extracted from leaf of Suaeda hetroptera kitag, then the CMO ( choline monooxygenase) cDNA was amplified using the reverse transcriptase polymerase chain reaction ( RT-PCR) method and cloned into pMD-T-simple vector. The positive clones from the Blue/White Screen were sequenced. After confirming its validity, the CMO gene fragment was cloned into pBI121 vector. Double enzyme restriction and PCR analysis indicated that the pBI121/CMO recombinant plasmid was successfully constructed.

OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Association of lumbar facet tropism and orientation with lumbar disc herniation is an issue of concern.OBJECTIVE: To overview the association of lumbar facet tropism and orientation with lumbar disc herniation and to clarify the existing problems.METHODS: The articles related to the association of lumbar facet tropism and orientation with lumbar disc herniation published between January 1967 and January 2017 were retrieved from CNKI and PubMed databases by computer. The keywords were "lumbar facet joints, lumbar disc herniation, facet orientation and tropism" in Chinese and English, respectively. Totally 42 articles were enrolled based on the inclusion and exclusion criteria.RESULTS AND CONCLUSION: (1) The association of lumbar facet tropism and orientation with lumbar disc herniation remains unclear. The relationship between the orientation of lumbar disc herniation and lumbar facet tropism is also controversial. (2) The main problems to drive controversy are: inappropriate control group and lacking normal lumbar facet tropism and orientation data; the large age range of subjects;the uncertain standard of measuring lumbar facet tropism.

To determine whether lentinan could upregulate the expression of human-beta-defensin-2(HBD-2) in pulmonary epithelial cells (SPC-A-1), we stimulated pulmonary epithelial cells with lentinan and detected the expression of HBD-2mRNA by RT-PCR test. The results demonstrated that the expression of HBD-2mRNA in SPC-A-1 could be induced by lentinan in a concentration and time-dependent manner.
Adjuvants, Immunologic ; pharmacology ; Epithelial Cells ; metabolism ; Humans ; Lentinan ; pharmacology ; Lung ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; beta-Defensins ; genetics ; metabolism

AIMTo observe the effects of sinomenine on the immune functions and apoptosis of murine lymphocyte as well as on human synovial fibroblast proliferation.

METHODSBoth in vivo and in vitro tests were adopted. The lymphocyte proliferation induced by mitogens was assayed by MTT method. Spleen T lymphocyte subtypes were tested with flow cytometry. Spleen lymphocyte apoptosis was analyzed by flow cytometry and DNA ladder methods. In vitro test was adopted to observe the effects of sinomenine on the proliferation of human fibroblast of rheumatoid arthritis.

RESULTSSinomenine can inhibit the proliferation of mouse lymphocytes induced by ConA, LPS and anti-CD3 mAb but not PMA in vitro, and inhibit the proliferation induced by LPS and PMA in vivo. Sinomenine can reduce up-regulated CD4+/CD8+ ratio of T lymphocyte subtype in adjuvant arthritis rat. At the same concentration increased apoptosis ratio. As to human synovial fibroblast, sinomenine can significantly inhibit proliferation of human fibroblast.

CONCLUSIONSinomenine can inhibit the immunological function and correct imbalance of CD4+/CD8+ ratio of T lymphocyte subtype. It can also increase apoptosis ratio of spleen lymphocyte. This may be the mechanism of its immunological inhibitory effect.

Animals ; Apoptosis ; drug effects ; Arthritis, Experimental ; immunology ; pathology ; Arthritis, Rheumatoid ; pathology ; CD4-CD8 Ratio ; Cell Proliferation ; drug effects ; Humans ; Lymphocytes ; cytology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Morphinans ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sinomenium ; chemistry ; Spleen ; cytology ; Synovial Membrane ; pathology

Objective To explore the relationship between gene polymorphism of neuropeptide Y (NPY) promoter and cerebral stroke subtypes according to TOAST (Trail of ORG 10172 in Acute Stroke Treatment) criteria in Chinese Han population. Methods The gene polymorphisms at position of-399T/C, -883Tgins/del and -602G/T in NPY promoter were detected by PCR method and gene sequencing in 190 cases of large-artery atherosclerosis stroke (LAA), 260 cases of small-artery occlusion (SAO), 60 cases of cardioembolism stroke (CE), 29 cases of stroke of other demonstrated etiology (ODE),10 cases of stroke of other undemonstrated etiology (UE) and 423 healthy control subjects. The PCR products were directly sequenced. Multivariate logistic regression was performed to analyze the relationship between gene polymorphism of NPY promoter and cerebral stroke subtypes according to TOAST by removing the confounding variables. Results Significant differences in the frequency of genotype CC and allele C at position of-399T/C were noted between the patients with SAO and controls (P=0.046, P=0.010). Compared with the control group, patients with LAA and SAO were more likely having high level of uric acid, hypertension, diabetes mellitus and heard disease (P＜0.05). No statistic differences in the frequency of genotype DD and allele D at position of-883Tgins/del were noted between patients with SAO and controls (P=0.0605, P=0.155). Gene polymorphisms of-399T/C,-883Tgins/del and -602G/T did not associate with an increased risk of having LAA, CE, ODE and UE.Conclusions The gene polymorphisms of promoter in position of-399T/C gene maybe associate with the happening of SAO; allele C at the position of-399T/C may raise the risk of the disease. There is no relationship between the gene polymorphisms of promoter at position of-399T/C, -883Tgins/del, -602G/T and the patients with LAA, CE, ODE and UE. High level of uric acid, hypertension, diabetes mellitus and heard disease history are the risk factors of LAA and SAO.

BACKGROUND: Gastric intestinal metaplasia (GIM) is generally considered to be the main mucosal background for the development of gastric adenocarcinomas. Using linked color imaging (LCI), we noticed that the color pattern in areas of GIM was purple mixed with white on the epithelium with signs of mist that were detected by the non-magnifying LCI observation. We have termed this endoscopic finding "Purple in Mist" (PIM). The aim of this study was to investigate whether PIM could be a useful optical sign for predicting GIM. METHODS: We prospectively evaluated consecutive patients undergoing endoscopy for various indications. The endoscopist used the LCI system to carefully observe the gastric antrum, body and angulus. When a PIM was identified in the surface layer, targeted biopsies were subsequently taken from this part. If the suspected area had no PIM on the surface, targeted biopsies were also taken. RESULTS: Sixty-three consecutive patients were included in this study. The prevalence of intestinal metaplasia (IM) was 29/63 (46%). In PIM-positive patients, the prevalence of IM was 23/26 (89%). Of these patients, 146 biopsy specimens were included in this study. For the diagnosis of IM, compared to histological assessment, the LCI finding had an accuracy of 91.1% (95%CI: 86.5%-95.7%), a sensitivity of 89.8% (95%CI: 81.3%-98.3%), a specificity of 91.8% (95%CI: 86.3%-97.2%), a positive predictive value of 84.6% (95%CI: 74.8%-94.4%), and a negative predictive value of 94.7% (95%CI: 90.1%-99.2%). CONCLUSIONS: A positive PIM finding in a suspicious lesion on LCI would complement LCI diagnosis of possible IM because of the positive predictive value of PIM. PIM could be a novel endoscopic marker for IM. TRIAL REGISTRATION ClinicalTrials.gov, No. NCT03092414; https://clinicaltrials.gov/ct2/show/NCT03092414?id=NCT03092414&rank=1.
Adult ; Aged ; Female ; Gastric Mucosa ; diagnostic imaging ; Gastroscopy ; methods ; Humans ; Male ; Metaplasia ; diagnostic imaging ; Middle Aged ; Prospective Studies ; Stomach Neoplasms ; diagnostic imaging

Objective To investigate epidemiologieal characteristics of hypothyroidism and factors influencing its outcome in the areas with different iodine intakes.Methods An epidemiologic follow-up study of thyroid diseases was performed in Panshan (iodine deficient),Zhangwu [mild iodine deficiency before universal salt iodination (USI) and more than adequate iodine intakes after USI ] and Huanghua ( iodine excessive) in China.A total of 3 761 subjects were investigated in 1999,and 3 018 (80.2% ) of them were followed up in 2004.FT_4,FT_3,TSH,thyroid autoantibodies and urinary iodine concentration were determined and thyroid B ultrasound was performed in all participants.Results Five-year cumulative incidence of overt hypothyroidism was 0.23%,0.47% and 0.35% in Panshan,Zhangwu and Huanghua,respectively.Twenty patients with autoimmune- induced overt hypothyroidism did not comply with the physician's orders to take thyroxine.Thyroid function spontaneously turned to normal in 55% of them,turned to subclinical hypothyroidism in 20% and maintained overt hypothyroidism in 25%.Five-year cumulative incidence of subclinical hypothyroidism in Zhangwu (2.60%) and Huanghua (2.89%) were significantly higher than that in Panshan(0.23% ) (both P＜0.01 ).One hundred patients with subclinical hypothyroidism were followed up,and 5% of them developed overt hypothyroidism,66% turned to euthyroidism and 29% maintained subclinical hypothyroidism.Raised serum TSH (＞6 mU/L) in the original survey ( OR = 3.4),positive thyroid autoantibodies in the follow-up study ( OR = 5.3 ) and more than adequate iodine supplementation in the mild iodine-deficient population ( OR = 8.0) were risk factors influencing outcome of subclinical hypothyroidism.Conclusion More than adequate iodine supplementation among a population in the mildly iodine-deficient area may increase the incidence of overt and subolinical hypothyroidism, and also influence outcome of subclinical hypothyroidism.

Objective To analyze the relationship between different iodine intakes and epidemiological features of hyperthyroidism in 3 communities with different iodine intake levels,as well as to explore the risk factor for the pathogenesis of hyperthyroidism.Methods A cohort study was performed in 3 Chinese rural communities with long-term stable iodine nutrition status defined as mild deficient (Panshan),more than adequate (Zhangwu, previously mild iodine deficiency and salt iodization lasting 8 years until 2004) and excessive (Huanghua).A total of 3 761 (934 males,2 827 females) inhabitants aged above 13 participated in the original investigation in 1999 and 3 018(709 males,2 309 females) of them received identical examinations after 5 years.Results Median urinary iodine concentration of school-aged children was 87.61zg/L in Panshan,213.9?g/L in Zhangwu and 633.5?/L in Huanghua (P＜0.05) at fallowing up,and no statistic significance was found when comparing the corresponding data from original survey (83.5?g/L,242.9?g/L and 650.9?g/L).The accumulative incidence of hyperthyroidism was 13.6/1 000,9.4/1 000 and 8.1/1 000 (P＞0.05),being 16.4/1 000,11.2/1 000 and 9.1/1 000 (P＞0.05) in female,respectively in Panshan,Zhangwu and Huanghua.In original healthy subjects (without history of thyroid disease and overt thyroid dysfunction) merely with either positive thyroid peroxidase antibody (TPOAb＞50 IU/ml) or thyroglobulin antibody (TgAb＞40 IU/ml),the accumulative incidence of newly diagnosed hyperthyroidism was not significantly different among 3 communities in follow-up survey.TSH＜0.3 mIU/L,positive TPOAb or goiter in original healthy participants were associated with the occurrence of overt hyperthyroidism in 5 years [ Logistic Regression,OR = 5.7 (95% CI 2.1-15 ),OR = 3.8 (95% CI 1.7-8.7 ) and OR = 3.1 (95% CI 1.4-6.8) respectively].Conclusion Iodine supplementation in mild iodine deficient area does not increase the incidence of overt hyperthyroidism ultimately.Long-term excessive iodine intake does not affect the incidence and pattern of hyperthyroidism.TSH＜0.3 mIU/L,positive TPOAb and goiter in healthy individuals are the risk factors for hyperthyroidism.

Objective To perform a follow-up survey about thyroid disorders in three rural communities with different iodine intakes in China,observe the incidences and natural outcomes of positive thyroid autoantibodies in euthyroid population,and also explore the influence of iodine intakes on these outcomes and autoantibodies.Methods In 1999,a cross-sectional study on thyroid disorders was performed in three rural communities of Pansan ( mild iodine deficient area),Zhangwu ( more than adequate iodine intake area) and Huanghua (excessive iodine intake area) in China.The 5-year follow-up study was performed in 2004.Both in 1999 and 2004,serum levels of TSH,thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb) were measured with the same method.Iodine in urine and B ultrasound on thyroid were also examined.Results Among the euthyroid subjects with normal TSH level in 1999,an increased prevalence of positive TPOAb in Zhangwu and an increased prevalence of positive TgAb in Hnanghua were observed in 2004 (both P＜0.05 ).Most of euthyroid subjects with positive TPOAb or positive TgAb in 1999 remained positive thyroid antibodies in 2004. The percentage was even higher in those with high levels of antibodies.Thyroid dysfunction was more frequent in subjects with positive TPOAb and/or TgAh than in those without thyroid antibodies (14.44% vs 3.11%,P＜0.01).For those with positive antibodies in 1999,the incidence of hypothyroidism in 2004 was 1.32%,8.46% and 15.38% in Pansan,Zhangwu and Huanghua,respectively (P＜0.05).The 5-year cumulative incidences of positive TPOAb (≥50 U/ml) and TgAb (≥40 U/ml) were 2.81% and 3.82%,respectively.The incidence of positive TPOAb was the highest in Zhangwu ( 3.84% ).The incidence of positive TgAb was the highest in Huanghua (5.07%),significantly higher than those in other two areas (P＜0.05).Conclusion The incidence of thyroid dysfunction is higher in subjects with positive antibodies than that in those with negative antibodies.The high iodine intake is a risk factor of hypothyroidism in subjects with positive thyroid antibodies.Sustained excessive iodine intake increases the incidence of positive thyroid antibodies.