Objective To clone the human Islet neogenesis associated protein(rhINGAP)gene,express the gene extraeellulary in Pichia yeast for.further study on biological function and animal test on INGAP.Methods INGAP gene Was amplified with PCR and inserted into the recombinant plasmidα/pUC18.Then,the fusion gene of α and INGAP was digested and inserted into the expression plasmid pPIC9K.The positive recombinant plasmid which integrated INGAP Was confirmed by restriction enzyme digestion and sequencing,and it Was linearized with Sal Ⅰ digestion and transfered into the yeast host strain GS115 through electroporation.The yeast transformants that harbor the desired gene INGAP with high copy were selected by the auxotroph mediam G418,and verified by PCR.The condition of hake-flask culture was optimized,and the recombinant human INGAP Was induced expression with methanol as the only Carbone source.The antigen activity of the desired protein Was detected by Western blotting and ELISA method.Results Recombinant plasmid αINGAP/pPIC9K were successfully constructed and three positive Pichia yeast transformants were obtained.The expressed protein had satisfactory antigen activity,which Was confirmed by the Western blotting and ELISA method.Conclusions Pichia yeast expressing human Islet neogenesis associated protein (rhINGAP)gene was successfully constructed.

OBJECTIVETo explore the scientificity of Yunqi theory based on incidence of disease related recognitions such as year evolutive phase, host climatic qi, six climatic factors, celestial manager qi, qi in the earth by analyzing data of hypertension incidence.

METHODSA total of 1,265 inpatients with confirmed first diagnosis as hypertension were recruited at Dongzhimen Hospital, Beijing University of Chinese Medicine from February 4, 1999 to February 4, 2011. Each period (including year evolutive phase, host climatic qi, guest climatic qi) was deducted based on Yun-qi theory. Their ratios of different phases and qi in the total number of hypertension inpatients number were calculated and statistically tested.

RESULTSThere was statistical difference in ratios at year evolutive phase, celestial manager qi, and qi in the earth (P < 0.05).

CONCLUSIONThe incidence of hypertension has certain difference in each period, which is consistent with Yun-qi theory.

Beijing ; epidemiology ; Humans ; Hypertension ; epidemiology ; Incidence ; Medicine, Chinese Traditional ; Qi ; Research

Objective To identify biomarkers in cerebrospinal fluid (CSF) by proteomic technology and develop a diagnostic model for neuropsychiatric lupus (NPSLE).Methods CSF proteomic spectra of 27 patients with NPSLE before and after treatment,and 27 controls including 17 patients with scoliosis,and 10 SLE patients without neuropsychiatric manifestation (non-NPSLE) were generated by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) combined with weak cationic exchange (WCX) magnetic beads.Data were analyzed with t test,non-parametric Kruskal-Wallis H test or Wilcoxon sign-rank test.A decision tree model for NPSLE classification was built based on the discriminating peaks.In addition,CSF samples of 12 patients with NPSLE,12 patients with lumbar disc herniation and 9 patients with other neurological conditions were employed as blind test group to verify the accuracy of the model.Results Twelve discriminating mass-to-charge (m/z) peaks were identified between NPSLE and controls.The diagnostic decision tree model,built with a panel of m/z peaks 8595,7170,7661,7740 and 5806,recognized NPSLE with the sensitivity and specificity of 92.6％ and 92.6％ based on training group samples,91.7％ and 85.7％ based on blind test group,respectively.Conclusion Potential CSF NPSLE biomarkers are identified by proteomic technology,the novel diagnostic model is sensitive and relatively specific for the diagnosis of NPSLE.

OBJECTIVETo investigate the therapeutic effect of Triamcinolone Acetonide and Pingyangmycin on lymphatic malformations in oral and maxillofacial regions.

METHODS29 patients with lymphatic malformations in oral and maxillofacial regions were divided into two groups to receive intra-lesion injection with Triamcinolone Acetonide and Pingyangmycin in experimental group, or with Pingyangmycin only in control group. The lesions involution and facial appearance were observed.

RESULTS2 years after treatment, the volume of small cyst and micro-cyst type mass shrank to (3.7 +/- 0.3)% and (4.2 +/- 0.4)%, respectively in experimental group, while (15.4 +/- 1.3)% and (24.1 +/- 3.1)% in control group. The lesion involution was markedly obvious in experimental group. Compare with control group, the facial asymmetry was greatly improved in experimental group.

CONCLUSIONSIntra-lesion injection with Triamcinolone Acetonide and Pingyangmycin is an effective method for the treatment of lymphatic malformations in oral maxillofacial regions. The mass can be shrank markedly to improve facial symmetry.

Adolescent ; Bleomycin ; analogs & derivatives ; therapeutic use ; Child ; Child, Preschool ; Drug Therapy, Combination ; Female ; Humans ; Infant ; Infant, Newborn ; Lymphatic Abnormalities ; drug therapy ; Male ; Maxillofacial Abnormalities ; drug therapy ; Treatment Outcome ; Triamcinolone Acetonide ; therapeutic use ; Young Adult

OBJECTIVETo explore the therapeutic effect of auricular therapy combined with optimized Yinxieling Formula on psoriasis vulgaris.

METHODSA randomized controlled single-blind clinical trial on 84 outpatients with psoriasis vulgaris was conducted. The patients were randomized to a treatment group (43 cases treated by auricular therapy combined with optimized Yinxieling Formula) and a control group (41 cases treated by optimized Yinxieling Formula alone) according to a random number generated by SPSS 17.0 software. The treatment duration for both groups was 8 weeks. The therapeutic effect was comprehensively measured by the primary outcome measure [Psoriasis Area and Severity Index (PASI) reduction rate] and the secondary outcome measure [PASI, Visual Analogue Scale (VAS), Dermatology Life Quality Index (DLQI), Self-rating Depression Scale (SDS), and Self-rating Anxiety Scale (SAS)]. The outcomes of both groups were obtained and compared before and after the intervention.

RESULTSThe PASI reduction rate in the treatment group was 74.4% (32/43), which was higher than that in the control group (36.6%, 15/41, P<0.01). The PASI scores decreased in both groups after treatment and was lower in the treatment group compared with the control group P<0.01). With stratified analysis, there were significant differences between the PASI scores in the following subgroups: age 18-30, baseline PASI>10 and stable stage (P<0.05). DLQI decreased in both groups on some categories after treatment, but there were no significant differences between the two groups in SDS, SAS and VAS (P >0.05). No obvious adverse reactions were found in either group.

CONCLUSIONThe therapeutic effect of auricular therapy combined with Optimized Yinxieling Formula was superior to Optimized Yinxieling Formula alone with no obvious adverse reaction.

Acupuncture, Ear ; Adult ; Anxiety ; complications ; Combined Modality Therapy ; Depression ; complications ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Male ; Outpatients ; Pain Measurement ; Psoriasis ; complications ; drug therapy ; Quality of Life ; Severity of Illness Index ; Single-Blind Method

Objective: To explore whether the pro-inflammatory effect of endoplasmic reticulum stress in placental tissues involves in the genesis of gestational diabetes mellitus (GDM). Methods: Forty gravidas who underwent regular prenatal examinations and delivered at Tongji Hospital were recruited from January to December, 2016. Among them, 20 were GDM women (GDM group), and the remaining twenty were served as the control, which were selected from those without GDM and matched for age and gestational weeks to the GDM group. Placental tissues were collected from the two groups. The ultrastructure of endoplasmic reticulum in trophoblast cells was observed under transmission electron microscope. The expression of glucose-regulated protein-78 (GRP-78), a marker protein for endoplasmic reticulum stress, and C/EBP homologous protein (CHOP) were detected using Western blotting. Five placental tissue samples were collected from normal gravidas for explant culture. Three subgroups were set up according to different culturing methods including culturing with IL-1β (5 ng/ml) for 20 h (IL-1β model group), 30 μmol/L thapsigargin (TG, an endoplasmic reticulum stress agonist) for 2 h after treating with IL-1β (5 ng/ml) for 18 h (IL-1β+TG intervention group) or with no stimulation (blank control group). Western blotting was used to detect the expressions of GRP-78, CHOP and glucose transporter 4 (GLUT4) in placenta explants. The mRNA expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Statistical analysis was performed using one-way analysis of variance, LSD and t test. Results: (1) In the GDM group, increased number and size of endoplasmic reticulum cisternae were observed in trophoblast cells. Moreover, obviously dilated endoplasmic reticulum and different size of fragments and vesicles were also seen under electron microscope. While the endoplasmic reticulum in the placental tissues of the control group showed no obvious swelling. (2) The expression of GRP-78 and CHOP protein in the GDM group were higher than those in the control group (0.90±0.17 vs 0.48±0.08, t=2.24; 0.85±0.13 vs 0.46±0.12, t=2.10; both P<0.05). (3) Compared with the blank control group, the expression of GRP-78 and CHOP protein in the IL-1β model group increased significantly (0.87±0.18 vs 0.36±0.07, t=2.67; 1.14±0.09 vs 0.78±0.06, t=3.20; both P<0.05); but the expression of GLUT4 protein significantly decreased (1.00±0.14 vs 2.21±0.49, t=2.40, P<0.05); the expressions of IL-6 and TNF-α mRNA significantly increased (0.89±0.23 vs 0.30±0.06, t=2.31; 0.62±0.16 vs 0.17±0.09, t=2.29; both P<0.05). Compared with the IL-1β model group, the expression of GRP-78 and CHOP protein significantly increased in IL-1β+TG group (2.02±0.32 vs 0.87±0.18, t=3.11; 2.18±0.31 vs 1.14±0.09, t=3.16; both P<0.05); the expression of GLUT4 protein significantly decreased (0.39±0.19 vs 1.00±0.14, t=2.66, P<0.05); the expression of IL-6 and TNF-α mRNA increased significantly (1.67±0.25 vs 0.89±0.23, t=2.26; 1.42±0.27 vs 0.62±0.16, t=2.51; both P<0.05). Conclusions Endoplasmic reticulum stress may be associated with increased release of pro-inflammatory cytokines in placental tissues of some GDM women and involved in the onset and development of GDM.

Objective To report the first family of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy(CARASIL)in China,and to explore its clinicopathological characteristics.Methods The clinical,imaging and pathological findings of the two patients were studied, and the sequence of the exons from 2 to 6 on Notch 3 in the chromosome 19 was detected.Results Two siblings were born from consanguineous parents.The ages at onset were 25 and 20 years old,respectively. Clinically,both of the patients were characterized by alopecia,acute lumbago,progressive intellectual deterioration,ataxia,pseudobulbar palsy and pyramidal tract signs.MRI demonstrated diffuse leucoencephalopathy and multiple subcortical infarcts on both hemisphere.The sural nerve biopsy on the elder sister demonstrated concentric thickening of vascular wall,narrowing of the lumen and mild fibrous proliferation of the intima.There were no amyloid,PAS granular deposition and uhrastructural granular osmiophilic material on the vascular wall.No mutation of exons from 2 to 6 on Notch 3 in the chromosome 19 was found by direct sequence.Conclusion The clinicopathological findings of the two patients fulfill the diagnostic criteria based on Fukutake.

OBJECTIVETo investigate relationship between expression of inducible nitric oxide synthase (iNOS) and proliferative potency of endothelium in hemangioma (HM).

METHODSImmunohistochemical staining was used to detect expression of iNOS and Ki-67 protein in 49 cases of HM and 29 cases of vascular malformation (VM).

RESULTSExpressive rate of iNOS and Ki-67 protein of 49 cases of HM was 38% and (10.98 +/- 7.93)%. Expressive rate of iNOS and Ki-67 protein of 29 cases of VM was respectively 3% and (0.03 +/- 0.19)%. The expressive rate of iNOS and Ki-67 protein of HM was significantly higher than that of VM (P = 0.001 and 0). The expressive rate of Ki-67 protein of HM of proliferative phase was (12.67 +/- 7.65)% , which was significantly higher than that of HM of extinctive phase, (7.27 +/- 7.49)% (P = 0.028).

CONCLUSIONSExpression of iNOS and Ki-67 protein of HM is significantly higher than that of VM, and the proliferative potency of HM is significantly higher than that of VM.

Child, Preschool ; Endothelium ; metabolism ; Endothelium, Vascular ; cytology ; Female ; Hemangioma ; metabolism ; pathology ; Humans ; Hyperplasia ; Infant ; Ki-67 Antigen ; metabolism ; Male ; Neovascularization, Pathologic ; metabolism ; Nitric Oxide Synthase Type II ; metabolism

OBJECTIVETo develop a best method of constructing influenza NP fusion gene containing enhanced green fluorescent protein (EGFP).

METHODSThe full-length NP gene of influenza A was amplified by RT-PCR and was inserted into an eukaryotic expression vector pEGFP-N1 in order to construct a fusion gene of pEGFP-N1-NP using three different methods. Method one, NP gene containing restriction endonucleases and pEGFP-N1 were both digested using the same restrict enzymes and ligated, yielding the fusion gene of pEGFP-N1-NP. Method two, NP gene was cloned into pMD19-T Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP; Method three, NP gene containing restriction endonucleases was cloned into pMD19-T Simple Vector to construct a plasmid of pMD19-T-NP. The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP.

RESULTSThe fusion gene of recombinant eukaryotic expression vector pEGFP-N1-NP was successfully constructed by using method three.

CONCLUSIONSThe full-length NP gene is obtained and its fusion gene of recombinant eukaryotic expression plasmid is successfully constructed. This study provides foundation for further understanding the biological function of NP protein and the mechanism of diseases induced by influenza A virus.

Artificial Gene Fusion ; Cloning, Molecular ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Polymerase Chain Reaction ; RNA-Binding Proteins ; genetics ; Viral Core Proteins ; genetics

Channel syndrome differentiation is a more commonly-used syndrome differentiation method of Professor HE Pu-ren clinically, which includes the 3 aspects: differentiation of diseases and syndromes on the channel parts along the body surface; differentiation of diseases and syndromes of the internal organs connected with the channels; differentiation of qi and blood of the channels. According to results of the channel syndrome differentiation, with flexible application of the HE's Santong methods and selection of corresponding treatment program, many complicated and difficult diseases are cured.
Acupuncture Therapy ; methods ; Diagnosis, Differential ; Humans ; Medicine, Chinese Traditional