ObjectiveTo observe the microstructure and ultrastructure of platelet-rich plasma gel. MethodsPRP gel samples were obtained by two-step centrifugation. The platelets were counted before and after centrifugation. TGF-β1, PDGF-AB were measured in the PRP gel and the whole blood using the enzyme-linked immunosorbent assay. PRP gel samples were observed with macroscopic observation, HE staining, transmission (TEM) and scanning electron microscopy (SEM). ResultsThe platelet concentration of PRP was 458% of whole blood. TGF-β1, PDGF-AB were found in high concentrations in PRP gel samples. Both SEM and TEM showed that PRP gel mainly contained fibrillar material with striated band similar to fibrin filaments, and platelet. ConclusionPRP gel may be an ideal injectable scaffold material for constructing tissue engineering nucleus pulposus.

OBJECTIVE：To identify two leguminosae Chinese drugs,semen phaseoli,semen cassiae,and their adulterants by electrophoresis and to study the dyeing effect of coomassie brilliant blue G-250.METHOD:Gel electrophoresis of soluble protein was used.RESULTS:The elect rophoretograms of semen phaseoli,semen cassiae and their adulterants are used d ifferent.CONCLUSION:The electrophoretograms can be used to different iate semen phaseoli and semen cassiae from their adulterants.Coomassie brilliant blue G-250 can be used in the electrophoresis identification.

Objective To prepare the standard molecular weight fragment mixtures. Methods Primers were designed to prepare clones which contained different sizes of standard molecular weight fragments. The template used for amplification of insert fragments was the pMD18-T vector. Bacteria culture and plasmid extraction were used to obtain abundant target fragment. Unlabeled DNA fragments were prepared by double digestion of the recombinant plasmids, and the fluorescent adaptor was prepared by annealing with two partial reverse complimentary DNA fragments. The unlabeled fragments and fluorescent adaptor were connected by DNA ligation reaction assisted with T4 DNA ligase. In this way, different sizes of standard molecular weight fragments were prepared. Standard molecular weight fragment mixture was finally prepared by mixing all the fragments together before purification. Results Ten standard molecular weight fragments of different sizes were prepared. The sizes of each fragment are 80bp, 124bp, 194bp, 224bp, 254bp, 304bp, 349bp, 399bp, 424bp and 454bp. The internal standard could accurately determine the size of PCR products amplified with the DNATyper15 kit. Conclusion Using this method, the standard molecular weight fragment mixture which meet the requirements of research and laboratory use was prepared, perfectly providing a new method for preparation of the DNA molecular weight standards. The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.

Objective To observe the efficacy of Tongmai Acupoint Plaster combined with Alprostadil injection for treatment of lower extremity arteriosclerosis obliterans (ASO). Methods Totally 180 cases of ASO Ⅱ(intermittent claudication period) patients were randomly divided into acupoint sticking group, combination group, Western medicine group, with 60 cases in each group. All groups quit smoking, had diet control, proper exercise, control of blood sugar, stop hypolipidemic and anti-coagulation medicine. The acupoint sticking group was treated with Tongmai Acupoint Plaster, each selected with the efficacy of treatment of lower limbs paralysis acupoint 10-12, alternating dressing; dressing was changed 1 time a day, 40 day as a treatment course. Western medicine group was given Alprostadil injection 10 μg, with 10 mL of normal saline, intravenous infusion, once a day, 15 d as a treatment course, for two courses, with 10 d as interval. Combination group was treated with Tongmai Acupoint Plaster combined with Alprostadil injection, the same as the application method above. The clinical efficacy, the scores of symptoms and signs, the ankle brachial index, blood lipid, the peak value of blood flow in the tibial anterior,posterior tibial, and dorsalis pedis arteries were observed and compared between the 2 groups. Results The total effective rate was 78.33% (47/60) in the acupoint sticking group, 80.00% (48/60) in the Western medicine group, and 93.33%(56/60) in the combination group, with the combination group better than the acupoint sticking group and Western medicine group (P<0.05). There was no statistical significance between acupoint sticking group and Western medicine group (P>0.05). There was statistical significance in scores of symptoms and signs, ankle brachial index before and after treatment in the three groups (P<0.05, P<0.01); There was no statistical significance after treatment (P>0.05). There was statistical significance in blood lipids before and after treatment in acupoint sticking group and combination group (P<0.05). There was statistical significance in tibial anterior, posterior tibial, and dorsalis pedis artery peak before and after treatment in the three groups (P<0.05, P<0.01). Conclusion Tongmai Acupoint Plaster has good efficacy in the treatment of ASO, equivalent with the efficacy of Alprostadil injection, which can regulate blood lipids and improve arterial blood flow. Ther combination of them has better efficacy.

The pathogenesis of colorectal cancer is related to genetic and environmental factors. Chronic inflammation of the intestinal mucosa is one of the most important factors in environmental factors. In inflammation factors, Intestinal flora plays a role in bridging and inducing intervention. Environmental changes disrupt the homeostasis of intestinal flora, intestinal flora maladjustment occurred, bacteria induces intestinal mucosal inflammation. The pathogenic bacterium adheres to the surface of the intestinal mucosa, it produces cytotoxic and genotoxic products, intestinal epithelial cells undergo genetic damage. The synthesis and metabolites of bacteria also control the occurrence of colorectal tumor process. This process leads to the progression of inflammation to cancer. This article reviews the process of intestinal flora mediated from inflammation to carcinogenesis, and the latest progress in related pathogenic bacteria, and proposes that the intestinal flora can be adjusted and targeted removal of the conditional pathogenic bacteria, and achieve the goal of cancer prevention. The relationship between intestinal flora, intestinal inflammation and colorectal cancer is reviewed.

Objective To observe the application of multiple fluorescent PCR (Polymerase Chain Reaction) in the diagnosis and clinical detection of bloodstream infection. Methods 256 blood cultures were collected by the Laboratory Department of Yinzhou People′s Hospital from January 2018 to May 2018, and were detected by multiplex fluorescent PCR. The results of the PCR were compared with the traditional blood culture bacteria identification instrument (traditional blood culture method). The number of positive and negative samples and the number of corresponding samples of the two methods were counted. Then, they analyzed the specificity and sensitivity of multiplex fluorescence PCR in the diagnosis of bloodstream flow infections. Results A total of 18 pathogenic microbes are detected through blood culture and PCR. Multiple fluorescent PCR detects 142 positive samples and 114 negative samples. Among them, 132 samples also show positive through blood culture, and 111 samples show negative. The consistency rate between multiple PCR and traditional blood cultures is 91.8％ (235/256). The negative prediction rate of PCR is 97.4％ (111/114), sensitivity rate 97.8％ (132/135), specificity rate 91.7％ (111/121). 10 samples show positive through multiple fluorescence PCR but negative for blood culture, 3 samples show positive through blood culture but negative for PCR. Besides, there are 3 types of pathogens that exceed the detection range of PCR. Conclusions Multiplex PCR method can detect 17 pathogens in blood culture specimens of patients, which can not only optimize the traditional blood culture process, but also greatly shorten the reporting time and improve the detection rate of blood culture methods. Especially for patients treated with antibiotics, it can reduce missed detection and improve the diagnostic rate of bloodstream infections.

Objective To investigate the characteristics and rules of tongue image in patients with chronic fatigue syndrome(CFS)with different fatigue degree.Methods Totally 917 patients with severe chronic fatigue syndrome(severe CFS group),351 patients with mild chronic fatigue syndrome(mild CFS group)and 1216 healthy controls(healthy control group)were enrolled in the physical examination center of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine.The tongue image data of subjects in the three groups were collected using TFDA-1 digital tongue and face diagnostic instrument,and the color space indexes of RGB,HSI,Lab and YCrCb were used to analyze the tongue image differences of CFS people with different fatigue degrees and the tongue image features of CFS patients with liver-qi stagnation syndrome,damp-heat stasis syndrome and spleen deficiency syndrome.Results Compared with the healthy control group,the tongue image indexes TB-R,TB-G,TB-B,TB-I,TB-L,TB-Y,TC-H,TC-I,TC-L and TC-Y increased in the severe CFS group;TB-S,TB-a,TC-S,TC-a,TC-Cr decreased(P<0.05).TB-R,TB-G,TB-B,TB-I,TB-L,TB-Y,TC-R,TC-G,TC-B,TC-I,TC-L and TC-Y increased in severe CFS group compared with mild CFS group.TB-H and TB-b increased in mild CFS group compared with healthy control group.The comparison of syndromes in severe CFS group showed that TB-a,TB-Cr,TC-S,TC-a,TC-Cr and TB-S increased in liver-qi stagnation syndrome compared to damp-heat stasis syndrome;TB-G,TB-B,TB-I,TB-L,TB-Y,TB-b,TB-Cb,TC-G,TC-B,TC-H,TC-I,TC-L,TC-Y and perAll decreased(P<0.05).Compared with spleen deficiency syndrome,TB-a,TB-Cr,TB-CON,TB-ENT,TB-MEAN,TC-a,TC-Cr,TC-CON,TC-ENT,TC-MEAN increased in liver-qi stagnation syndrome;TB-ASM,TC-S and TC-ASM decreased(P<0.05).Compared with spleen deficiency syndrome,TB-a,TB-b,TB-Cr,TB-Cb,TB-CON,TB-ENT,TB-MEAN,TC-G,TC-B,TC-H,TC-I,TC-L,TC-a,TC-Y,TC-Cr,TC-CON,TC-ENT,TC-MEAN,perAll increased;TB-ASM,TC-S and TC-ASM decreased(P<0.05).The comparison of mild CFS syndrome showed that there was no statistical significance between liver-qi stagnation syndrome and spleen deficiency syndrome(P>0.05).TB-Cr,TC-a,TC-Cr and perAll increased and TC-S decreased in damp-heat stasis syndrome compared with spleen deficiency syndrome(P<0.05).TB-S,TB-a,TB-Cr,TC-S,TC-a,TC-Cr increased,and TB-G,TB-B,TB-I,TB-Cb,TB-b,TC-b and TC-Cb decreased(P<0.05)in liver-qi stagnation syndrome compared with damp-heat syndrome.The distribution trend of TC-S was as follows:dampness-heat syndrome

Objective:To serologically and genotypically analyze the pedigree of a case with a new allele c.175_176insGA of B el subtype and preliminarily explore the molecular mechanism of weak expression of glycosyltransferase B. Method:In the descriptive study,a 23-year-old male voluntary blood donor and his family members were selected for the study. The ABO and Le blood types of the proband and his family members was identified by the test tube method. The agglutination inhibition test was applied to detect the B and H antigens in saliva, and the Sanger sequencing and PacBio (Pacific Bioscience) third-generation haplotype sequencing were performed on the study subjects to identify genotypes. Finally, Expasy software were applied to amino acid translation of DNA sequences and prediction of protein length after gene alteration. ORF finder was applied to predict alternative start codons as well as open reading frames of mRNA, and protein expression mechanisms were analyzed.Results:The proband and her sister were B el subtype, her mother was AB el subtype, her father was normal O type, and all members of the family were Le(a+b+) phenotype. Sanger sequencing results showed that a new allele of c.175_176insGA was found in exon 4 of the proband, her mother, and her sister. Third-generation haplotype sequencing detected the haplotypes of the family members, which revealed that the proband was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA), the father was ABO*O.01.02/ABO*O.01.02, the mother was ABO*A1.02/ABO*BEL.NEW (c.175_176insGA), and the sister was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA). Analysis of the protein expression mechanism indicated that although the new allele of ABO*BEL.NEW was presumed to cause a frameshift mutation and result in a premature stop codon p.Asp59Glu*fs20 in exon 5, encoding an inactive glycosyltransferase, an alternative start codon could be utilized to initiate translation of B el subtype functional glycosyltransferase. Conclusion:Expression of the new allele of B el subtype is associated with the translation of B el subtype glycosyltransferase initiated by alternative start codons.

Peroxisome proliferator activated receptors (PPARs) are ligand activated nuclear transcription factors and one of the members of the non steroidal nuclear receptor superfamily.It can be divided into PPAR alpha,PPAR beta / delta and PPAR gamma three subtypes according to the different of its structure and function.Previous studies showed that PPARs participated in biochemical reactions and the regulation of other important biological activities such as lipogenesis,glucose metabolism,inflammation,insulin sensitivity and so on.Recent researches showed that PPARs also had effect of anti-fibrosis,protecting ischemia-reperfusion injury and inhibiting the growth and differentiation of tumor cells.This article reviewed the recent research progress of PPARs in these liver diseases.