Objective To study the curative effect of limited-acupotomy therapy on chronic lumbosacral osteo-fascial compartment syndrome. Methods 59 patients were randomly recruited into a control group (with 29 patients) and a treatment group (with 30 patients). The control group was treated with general-acupotomy therapy, and treatment group was treated with limited-acupotomy therapy. Evaluate the curative effects before the first and the second therapy, and 3 months after the therapy respectively, as well as VAS pain, JOA and CODI scores. Results The curative effect was 96.56% and 100% respectively in the control group and the treatment group 3 months after the treatment. The difference between the two groups was not statistically significant(χ2＝0.19,P＞ 0.05). As to VAS pain scores, JOA and CODI scores, the difference among the three stages of the treatment were significant (in control group F＝165.70, 99.90, 106.60 respectively, in treatment group F＝279.76, 154.34, 67.36 respectively, P＜0.01)in both groups respectively, but the difference between the two groups were not significant(P＞0.05) in each stage. Conclusion Limited-acupotomy therapy was safe and effective in treating chronic lumbosacral osteo-fascial compartment syndrome.

Purpose To evaluate the application value of ERCC2 gene polymorphism ( rs3916840 C/T, rs1799793 G/A and rs238416 G/A) detection in molecular pathological diagnosis of breast cancer. Methods The polymorphisms of ERCC2 ( rs3916840, rs1799793 and rs238416) were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay in 101 patients with breast cancer and in 101 cancer-free controls. Results Analysis of the data showed a 0. 287-fold increased risk of breast cancer due to the deletion of genotype GA at rs238416 (P<0. 001, 95%CI: 0. 153 ~0. 537). However, polymorphisms of rs3916840 and rs1799793 were not associated with breast cancer risk (P>0. 05). Furthermore, Heterozygous genotype of rs3916840 was significantly associated with tumor size (P=0. 049), heterozygous genotype of rs1799793 was significantly associated with PR sta-tus and triple negative breast cancer (P=0. 037). Remarkably, the genotype frequency of GA in p53-positive patients was lower than that in p53-negiative patients (P=0. 026). Conclusions These results indicate that the polymorphism of rs238416 of ERCC2 is sig-nificantly associated with breast cancer risk. Tumor size, PR status, triple negative breast cancer, and p53 protein expression are asso-ciated with polymorphisms of ERCC2 (rs3916840, rs1799793 and rs238416) respectively. ERCC2 gene polymorphism detection is useful for the early diagnosis and prognosis evaluation of breast cancer.

Objective To explore the effects of different extracts of modified Siwu Siteng Decoction (TFT-Ⅰ, TFT-Ⅱ) on rat models with hyperuricemia. Methods Rat model with hyperuricemia was induced by hypoxanthine and oxonic acid potassium salt. Seventy rats were randomly divided into 7 groups:normal group, model group, allopurinol group, TFT-Ⅰ high and low dose groups, TFT-Ⅱhigh and low dose groups. Each dose group was given lavage with related medicine, and blank group and model group were given lavage with distilled water for one week, respectively. Uric acid in serum (SUA) was measured through fully automatic biochemical analyzer, and xanthine oxidase (XOD) activity in serum and liver tissue was measured through colorimetric method. Results Compared with model group, TFT-Ⅰ and TFT-Ⅱ high dose groups significantly reduced the level of SUA, serum and liver XOD activity (P<0.05), with significant statistical difference between the two groups (P<0.05). TFT-Ⅰ and TFT-Ⅱ low dose groups had no significant effects on SUA, and XOD activity in serum and liver (P>0.05). Conclusion Alcohol soluble ingredients of astragalus membranaceus and Siwu decoction in modified Siwu Siteng Decoction can effectively reduce the level of blood uric acid.

Objective To investigate toxicity and teratogenic effect of Microula seed oil on embryo of rat.Methods 150 mature Wistar rats (100 females, 50 males) were selected with female∶ male = 2∶1 cage match. During the daily morning examination the sperm was discovered in the vagina as the zero day for conception.100 pregnant rats were randomly divided into 5 groups (n=20 in each group): treatment group (included 3 groups: to give Microula seed oil 2.5 g /kg, 5.0 g /kg, 10.0 g /kg body weight, ig, respectively), cyclophosphamide group(7 mg/ kg body weight, sc) and the edible oil group (to give dose, such as volume as Microula seed oil, ig). From the 7th day of pregnancy, the treatment group, the edible oil group were given intervention once a day for 10 days. From the 11th day of pregnancy cyclophosphamide group was given cyclophosphamide as intervention once a day, for 3 days. On the 20th day the pregnant rats were killed.Results The body weight of pregnant rats and the rate of live births were significantly higher in the Microula seed oil dose group than in the cyclophosphamide group (P<0.01), stillbirth rate and birth rate of absorption was significantly lower in the Microula seed oil dose group than in the cyclophosphamide group (P<0.01), and no significant difference from the edible oil group (P> 0.05); the fetal rat, body weight, body length, tail length in all groups of Microula seed oil was no significant difference from the edible oil group (P> 0.05). There was no malformation in appearance, viscera, bones in the treatment group and the edible oil group while there were 112 fetal rats with deformity in 140 in the cyclophosphamide group.Conclusion Microula seed oil at doses of 2.5 g /kg, 5.0 g /kg, 10.0 g /kg body weight had no significant role in the toxicity and teratogenicity on embryos of pregnant rats.

Objective To investigate the prevalence and severity degree of post traumatic stress disorder(PTSD), anxiety and depression, and to explore the different mental profile in different groups involved with an earthquake. Methods 26 days after 8.0 grade earthquake in Wenchuan, psychological rescue team of State Administration of Traditional Chinese Medicine went to some communities in Deyang and Mianzhu area of Sichuan province. In this period, a total of 119 survivors, including students(42), teacher(40) and masses(37), were investigated through impact of event scale (IES), self-rating anxiety scale (SAS) and self-rating depression scale(SDS). Results①The detection rate of score over 19 in IES in 3 groups (students, teacher and masses) were 69％, 80％ and 91.9％, respectively. There was significant difference among these 3 groups (F=5.611, P=0.005<0.01) in respect of severity degree of PTSD. ②The values of severity degree of anxiety in 3 groups were significant higher than normal value (P<0.001, all). There was also significant difference among these 3 groups (F=3.376, P=0.038<0.05) in respect of severity degree of anxiety, with the masses group being significant higher than student and teacher group (P=0.029, P=0.022, respectively). ③The values of severity degree of depression in these 3 groups were significant higher than normal value (P<0.05, all). But there was no significant difference (F= 0.670, P=0.514) among these 3 groups. ConclusionThe results suggested the importance to assess PTSD, anxiety and depression in post-disaster area, especially PTSD and anxiety evaluations in the masses group.

OBJECTIVETo fusionaly express the HAV 3a (located in 1403-1456 aa) and 3d located in 1719-1764 aa cDNA gene fragments in prokaryotic system; to investigate the antigenicity and application of recombinant protein.

METHODSBy using PCR technique, 3a and 3d gene fragments were cloned. Choosing pET-30a as the expressive vector, the recombinant plasmid Pet-3ad was constructed and pET-3ad was expressed in Escherichia coli after inducing by IPTG. By affinity chromatography, purified recombinant protein was obtained. By using Western blot analysis and indirect ELISA to detect its antigenic activity.

RESULTSRecombinant plasmid pET-3ad was proved to construct successfully by enzyme-digestion and sequence measurement. Recombinant protein P3ad(18,000) was obtained in BL21(DE3) and purified after Ni+ affinity chromatography. Western blot analysis and indirect ELISA showed that P3ad had specific antigenicity.

CONCLUSIONSRecombinant plasmid pET-3ad was proved to construct successfully by enzyme-digestion (Nco I/Hind III) and sequence measurement. Recombinant protein P3ad(18,000) was obtained in BL21(DE3) and purified after Ni+ affinity chromatography. Specific immunoblotting appeared at 18,000 by western blot analysis, which showed the recombinant protein P3ad had specific antigenicity, indirect ELISA further proved its antigenicity.

Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Hepatitis A virus ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; immunology

[Abstract] Objective: To explore the expression of PSME3 (proteasome activator complex subunit 3) in gastric cancer (GC) tissues and its correlation with the prognosis of GC patients, and to further analyze its effect and mechanism in the occurrence and development of GC. Methods: The expression level of PSME3 gene in GC tissues was analyzed with TCGA and UALCAN database. qPCR was used to verify the expression of PSME3 in GC tissues and corresponding adjacent normal tissues that resected from 40 GC patients who were surgically treated in the Fourth Hospital of Hebei Medical University from January 2017 to December 2018. ROC curve and KaplanMeier plotter method were used to analyze the value of PSME3 mainly in diagnosing and predicting the prognosis of GC patients. The biological processes and pathways that PSME3 involved in were further analyzed. Results: The expression level of PSME3 in GC tissues was significantly higher than that in normal tissues, and it’s high expression was significantly correlated with the tumor stage, pathological subtype, status of lymph node metastasis and Helicobacter pylori infection in GC patients (all P<0.01). PSME3 was also highly expressed in GC tissue samples collected by the qPCR confirmatory detection group (P<0.01). PSME3 could distinguish gastric cancer patients from normal people with an AUC value of 0.808. The overall survival time, the first progression survival time and post progression survival time of the GC patients with low PSME3 expression were longer than those in the patients with high PSME3 expression (all P<0.01). Mechanism research found that PSME3 mainly played an oncogenic role of the development of GC by regulating cell cycle, mTORC1 signaling pathway, PI3K/AKT/mTOR signaling pathway and TGF- β signaling pathway etc. Conclusion: PSME3 is highly expressed in GC tissues, and it is significantly related to the poor prognosis of GC patients. It plays an oncogenic role in the occurrence and development of GC.

The aim of this study was to screen the active regions and transcription factor binding sites in the promoter of the CBD103 gene related to Arctic fox coat color, and to provide a basis for revealing the molecular genetic mechanism of CBD103 gene regulating the coat color formation. The 5'-flanking region fragment 2 123 bp of Arctic fox CBD103 gene was cloned, and 4 truncated promoter reporter vectors of different lengths were constructed. The promoter activity was detected by the dual-luciferase reporter assay system. Point mutations were performed on the 3 predicted specificity protein 1 (Sp1) transcription factor binding sites in the highest promoter active region, and 3 mutant vectors were constructed. The activity was then detected by the dual-luciferase reporter assay system. The results showed that the region 1 656 (-1 604/+51) had the highest activity in the 4 truncated promoters of different lengths, and the promoter activity of the three mutant vectors constructed in this region were significantly lower than that of the wild type (fragment 1 656). The region of -1 604 /+51 was the core promoter region of CBD103 gene in Arctic fox and -1 552/-1 564, -1 439/-1 454 and -329/-339 regions were positive regulatory regions. This study successfully obtained the core promoter region and positive regulation regions of the Arctic fox CBD103 gene, which laid a foundation for further study on the molecular genetic mechanism of this gene regulating Arctic fox coat color.
Animals ; Binding Sites ; Foxes ; Luciferases ; Promoter Regions, Genetic ; Sp1 Transcription Factor ; beta-Defensins

ObjectiveTo investigate the pharmacodynamic characteristics and explore the molecular mechanism of Honghua oral liquid （HOL） in relieving neuropathic pain （NP）. MethodHealthy male SD rats were randomly assigned into sham group， model group， low-， medium-， high-dose （0.5， 1.0， 2.0 mL·kg^-1·d^-1， respectively） HOL groups， and a positive drug （pregabalin， 25 mg·kg^-1·d^-1） group， with 6 rats in each group. Spinal nerve ligation （SNL） of L5 was conducted in other groups except the sham group. Drug administration was performed 3 days after the SNL surgery for 2 consecutive weeks， and samples were collected after the end of the administration. During the treatment period， the mechanical pain threshold and cold pain threshold were determined to measure the pain-relieving effect of HOL. Transcriptome sequencing was performed on hippocampal tissue samples from the sham， model， and high-dose HOL groups， and differentially expressed genes between the sham group and the model group as well as the model group and HOL high-dose group were obtained. After pathway enrichment analysis， we selected the targets which were closely related to neuroinflammation for validation， and predicted the specific binding sites of the major active components in HOL with the targets through molecular docking. In addition， the serum levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） were determined by enzyme-linked immunosorbent assay （ELISA） to evaluate the effect of HOL on neuroinflammation in NP rats. ResultCompared with the sham group， SNL decreased the mechanical pain threshold and cold pain threshold （P<0.05）. Compared with the model group， HOL recovered the mechanical pain threshold and cold pain threshold （P<0.05）. The transcriptome data showed that 376 differentially expressed genes （DEGs） were identified between the model group and the sham group， including 124 upregulated genes and 252 downregulated genes， and 194 DEGs between the model group and the high-dose HOL group， including 33 upregulated genes and 161 downregulated genes. Among them， insulin-like growth factor 1（IGF1）， matrix metallopeptidase-2 （MMP-2）， matrix metallopeptidase-14 （MMP-14）， erb-B2 receptor tyrosine kinase 2 （ERBB2）， and integrin subunit alpha 5 （ITGA5） associated with NP were selected for further validation. The Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） results showed that compared with the sham group， the modeling up-gurelated the mRNA levels of the above five molecules in the hippocampus （P<0.01）. Compared with model group， HOL down-regulated the mRNA levels of these molecules （P<0.01）. The molecular docking results showed that the main active components of safflower， hydroxysafflor yellow A， kaempferol， and quercetin， formed stable hydrogen bonds with the amino acid residues of IGF1， MMP-2， MMP-14， ERBB2， and ITGA5. The enzyme-linked immunosorbent assay（ELISA） results showed that compared with those in the sham group， the serum levels of TNF-α and IL-10 were out of balance in the model rats （P<0.01）. Compared with the model group， HOL lowered the level of the pro-inflammatory cytokine TNF-α （P<0.01） and elevated that of the anti-inflammatory cytokine IL-10 （P<0.05）. ConclusionHOL exerts analgesic effect on SNL rats by inhibiting neuroinflammation.