AIM: To construct a human recombinant immunoglobulin library containing D-Dimer by using phage surface display technology. METHODS: Human immunoglobulin heavy chain and light chain genes were amplified respectively by RT-PCR from different human lymphocytes using family specific primers and signal sequences of immunoglobulin as half-nested PCR primers. The heavy chain and light chain PCR products were cloned into phagemid vector pComb3H and the human immunoglobulin recombination library was generated with helper phage VCSM13. RESULTS: A human combinatorial antibody library consisting of 2.8?10~8 in dependent clones was generated with a titer of 4.1?10~(17)PFU/L. The recombinant frequency of Fab genes was 46%. CONCLUSION: A human combinatorial antibody library was generated. It will be beneficial for selecting Fab antibodies of D dimer from the library.

[ ABSTRACT] AIM:To evaluate the clinical application of single nucleotide polymorphism array ( SNP array) in prenatal diagnosis for screening the abnormality of women with Down’ s syndrome ( DS) .METHODS:The amniotic fluid samples ( n=312) collected by amniocentesis for the DS screening abnormality women were tested by karyotyping and SNP array analysis, respectively.The findings of karyotyping and SNP array analysis were compared.RESULTS:Two cases of trisomy 21 were identified by karyotyping and SNP array analysis, but SNP array analysis failed to identify 6 cases of chro-mosome balanced structural rearrangement.SNP detected 176 cases copy number variants ( CNVs) in 303 cases normal karyotype were detected by SNP, including 106 benign CNVs, 61 variants of unknown significance (VOUS), 9 de novo CNVs, and none of them was pathogenic.The distribution difference of CNVs in DS screening positive group and DS screening positive plus advanced maternal age group was not statistically significant ( P>0.05) .Furthermore, we reported 14 kinds of CNVs for the first time in population.CONCLUSION:SNP array can further assure chromosome microdupli-cation/microdeletion.In normal karyotype fetus of prenatal diagnosis, SNP can detect some clinical significant CNVs.

Objective To explore the diagnostic value of five infection markers in bloodstream infection.Methods Randomly selected 110 bloodstream infection patients with positive blood cultures and 30 bacterial infection patients with negative blood cultures.Blood was simultaneously drawn with blood cultures;the complete blood count and C-reactive protein (CRP) levels were measured.The white blood cell count (WBC),neutrophil count (NEU),lymphocyte count (LMY),CRP level and neutrophil-lymphocyte count ratio (NLCR) were compared between the two groups.Results The levels of WBC,NEU,NLCR and CRP in bloodstream infection group were significantly higher than those in control group (P＜0.05),while LYM was significantly lower than that in control group (P＜ 0.05).Among these five infection markers,the area under the receiver operating characteristic curve (ROC-AUC) was the highest for NLCR (0.808) and LMY (0.756);when the cutoff value for NLCR was ＞9.33,sensitivity was 63.6％,specificity was 93.3％;and the cutoff value for LYM was ≤0.97,sensitivity was 58.2％,specificity was 86.7％.Furthermore,the NLCR of patients with gram-negative bloodstream infection was higher than those in patients with gram-positive bloodstream infection.NLCR showed important clinical significance in distinguishing strains of different bloodstream infections.Conclusions NLCR is the better predictors than routine parameters in diagnosing bloodstream infection.

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.

Objective To explore the difference in the regulating effect of electroacupuncture on serum insulin (Ins) and fat consumption between male and female simple obesity rat and the possible mechanism of its action.Methods A rat model of simple obesity was made by induction with sodium glutamate. The rats were randomized into model and electroacupuncture groups (male and female), 10 rats each. A normal group of 10 rats (male and female) was established as a control. Points Quchi, Zhongwan, Guanyuan, Housanli, Fenglong and Sanyinjiao were selected in the electroacupuncture group. Stimulation with a low frequency of 2 Hz was provided after needle insertion. The treatment was given once daily for 28 consecutive days. Serum Ins content, and greater omentum, pericardiac and perirenal fat weights were measured in the male and female groups of rats before and after electroacupuncture intervention. The comparisons were made.Results Serum Ins content, and greater omentum, pericardiac and perirenal fat weights were significantly lower in the electroacupuncture male and female groups of rats than in the model male and female groups of rats (P<0.01) and also in the electroacupuncture male group of rats than in the electroacupuncture female group of rats (P<0.05).Conclusions Electroacupuncture has different degrees of weight-reducing effect in both male and female obesity rats. The reducing effect on serum Ins content, and greater omentum, pericardiac and perirenal fat weights is better in male obesity rats.

PURPOSE: The purposes of the study were to examine psychological distress and health-related quality of life (HRQoL) in relocated and nonrelocated survivors aged 60 years and older, and to analyze predictors for psychological distress and HRQoL in older survivors 5 years after the 2008 Sichuan earthquake. METHODS: This was a cross-sectional descriptive study with 112 relocated older survivors and 156 nonrelocated older survivors. Our study used a multistage sampling method. The measurements used in the study included self-reporting questionnaire-20, medical outcomes study 36-item short form health survey, and an instrument measuring demographic and disaster-related characteristics. Descriptive and multiple linear regression analysis were performed to determine factors that contributed to psychological distress and HRQoL. RESULTS: The prevalence of psychological distress in relocated group (20.5%) was significantly higher compared to those in nonrelocated group (4.8%). Scores for HRQoL in relocated older survivors was significantly lower than those in nonrelocated older survivors. Relocation from preearthquake residence was the most significant predictor for psychological distress and HRQoL in the total sample. Other predictors were advanced age, lower educational level, the loss of family members during the earthquake, and the presence of chronic illnesses as well as the death of a spouse after the earthquake. CONCLUSIONS: Strategies can be designed in postdisaster recovery program, particularly for older survivors at high risk for psychological distress and poor HRQoL.
*Adaptation, Psychological ; Age Factors ; Aged ; Aged, 80 and over ; China/epidemiology ; Cross-Sectional Studies ; Disasters ; *Earthquakes ; Female ; *Health Status ; Humans ; Male ; Middle Aged ; Prevalence ; Quality of Life/*psychology ; *Residence Characteristics ; Socioeconomic Factors ; Stress, Psychological/*epidemiology ; Surveys and Questionnaires ; Survivors/*psychology

Objective To analyze composition principles of prescriptions for the treatment of diarrhoea caused by deficiency of spleen. Methods The prescriptions for diarrhoea caused by deficiency of spleen in the Prescriptions of Traditional Chinese Medicine Dictionary were collected, sorted and entered into the TCM Inheritance Support System (V1.3) to analyze the composition principles through the methods of entropy clustering and apriori. Results Based on the analysis of 1185 prescriptions and 815 medications, there were 33 medications with more than 50 frequencies. Composition principles were obtained through apriori method:50 herbal pairs were used with more than 50 frequencies, and 29 core combinations with more than 40 frequencies. Association principle rule were used for the analysis of those medications in the prescriptions (support≥20%, confidence≥0.9). Principles were obtained through entropy clustering:there were 19 core combinations which composed the new prescriptions, and 19 new prescriptions were found through hierarchical clustering method. There were 17 prescriptions matching with Sijunzi decoction (semblance=0.5). Conclusion Diarrhoea caused by deficiency of spleen should be treated with strengthening spleen and benefiting qi primarily, assisting with warming yang, excreting dampness and checking diarrhoea.

Objective To observe the dynamic changes of ghrelin of spleen-qi-deficiency rats and the intervention effects ofSijunzi Decoction.Methods Experimental rats were randomly divided into normal control group, model group andSijunzi Decoction group. Except for normal control group, spleen-qi-deficiency model was copied through the two-factor methods of breaking qi by bitter cold and swimming exhausted. Meanwhile,Sijunzi Decoction group was given 20 g/(kg?d)Sijunzi Decoction intervention. The activities of GAS, MTL, SS and VIP at different time points (14, 21, 28 d) in intestine and serum were detected by ELISA and RIA. At the same time the intervention effect of Sijunzi Decoction was studied.Results Compared with normal control group, GAS and MTL in intestine and serum of model rats decreased, while SS and VIP increased (P<0.05,P<0.01). Compared with model group, GAS and MTL in intestine and serum of rats in theSijunzi Decoction group increased, while SS and VIP decreased (P<0.05,P<0.01).Conclusion Ghrelin in intestine and serum of spleen-qi-deficiency rats shows dynamic coincidental changes.Sijunzi Decoction can treat spleen qi deficiency by regulating the activities of rat ghrelin.

Objective:To explore the clinical characteristics of autosomal dominant polycystic kidney disease (ADPKD).Methods:Clinical data of 103 patients with ADPKD first admitted to Renmin Hospital of Wuhan University from July 2017 to April 2021 were retrospectively analyzed. The clinical characteristics of patients in different renal function stages were analyzed, and multiple linear regression analysis was used to analyze the factors reflecting the severity of the disease.Results:Among the 103 patients with ADPKD, there were 49 males (47.6%), aged (51.23±10.99) years old. The extrarenal manifestation was mainly polycystic liver (64/71). The main clinical symptoms were gross hematuria (25 cases, 24.3%), lumbar distend and pain (37 cases, 35.9%) and hypertension (69 cases, 67.0%), appearing in the whole course of the disease. Early treatment was mainly drug conservative treatments (58 cases, 56.3%), followed by renal cyst aspiration (34 cases, 33.0%), and surgical treatments (11 cases, 10.7%). Patients in chronic kidney disease (CKD) stage 5 were mainly treated with conservative treatments (28/34). Laboratory examination results showed that hemoglobin, platelet, lymphocyte percentage and albumin in CKD stage 4-5 were lower than those in CKD stage 1-3 (all P<0.05) ; prothrombin time (PT), PT-international standardized ratio and plasma osmotic concentration in CKD stage 4-5 were higher than those in CKD stage 1-3 (all P<0.05). Multiple linear regression analysis showed that hemoglobin ( β=0.249, P=0.005), platelet ( β=0.207, P=0.005), lymphocyte percentage ( β=0.305, P<0.001) and plasma osmotic concentration ( β=-0.362, P<0.001) were correlated with estimated glomerular filtration rate. Conclusions:The clinical manifestations of ADPKD patients are hypertension, lumbar distend and pain, and gross hematuria, which can run through the whole stage of CKD. Polycystic liver is more common in extrarenal system. Hemoglobin, platelets, lymphocyte percentage and concentration osmotic concentration may be related to the disease progression of ADPKD.

Objective To investigate the prediction by intravascular ultrasound (IVUS) with monocyte to high density lipoprotein-cholesterol (HDL-C) ratio (MHR) of 12-month prognosis in patients with intermediate non-left main coronary lesions after percutaneous coronary intervention (PCI). Methods Patients with intermediate non-left main coronary lesions diagnosed by coronary angiography were tested of monocyte counts and HDL-C levels at admission with MHRs calculated. IVUS was used to examine plaque stability in target lesions. Patients were dviided into stable plaque group (n=44) and unstable plaque group (n=140) according to the IVUS results. PCI was then operated in patients with unstable plaque or with minimum lumen area<4 mm2. The major adverse cardiovascular events (MACE) were recorded during the follow-up period of 12 months after PCI. Results MHR was significantly higher in unstable plaque group than that in stable plaque group[(22.6±8.4) vs.(14.1±7.2),P<0.001]. Receiver-operating characteristic (ROC) analysis revealed that an MHR cut-off of 16.05 had 74.2％ sensitivity and 77.0％ specificity for prediction of 12-month MACE after PCI (AUC 0.78, 95％ CI 0.71–0.85, P<0.001). Besides, unstable plaque with MHR over 16.05 was an independent risk factor for 12-month MACE after PCI (adjusted HR 3.26, 95％ CI 2.48–4.14, P=0.020). Conclusions IVUS combined with MHR is a valuable index predicting the prognosiso f patients with intermediate non-left main coronary lesions who underwent PCI.