Background and purpose:Esophageal carcinoma is one of main malignancies with rapid course and a poor prognosis in China. The reasons of poor overall survival are the invasion and metastasis of the tumor. Matrix metalloproteinase (MMPs) play essential roles in promoting tumor invasion and metastasis. In this study, we aimed to investigate the expression and functional signiifcance of matrix metalloproteinase 16(MMP-16) in esophageal squamous cell carcinoma (ESCC). We expect to ifnd a lead molecule for the beneift of early detecting tumor and the development of novel treatment of ESCC. Methods:The expression levels of MMP-16 protein and mRNA in human ESCC and the matched normal tissues were determined by immunohistochemistry, Western blot and Real-Time PCR (RT-PCR). The stable Ec109 cell line with MMP-16 knockdown and negative controls were established by RNA interference technology. The cell migration, invasion, proliferation and cell apoptosis of MMP-16 in stable interfered Ec109 cell line was examined by cell counting, scratch test, Transwell test and lfow cytometry assays. The data were analyzed by t test. Results:MMP-16 protein was downregulated in cancerous group compared with the matched normal tissue and correlated with the clinical features of histological differentiation (P<0.05) and tumor stage (P<0.05). The levels of MMP-16 mRNA and protein in Ec109 were signiifcantly decreased by RNA intetrence (P<0.05). We demonstrated that MMP-16 silencing signiifcantly promoted cell invasion and migration (P<0.05), and inhibited cell apoptosis (P<0.05), while no significant effect was observed on cell proliferation (P>0.05). Conclusion: MMP-16 is downregulated in human ESCC tissues. The cell migration and invasion is promoted by interference of MMP-16 in Ec109, while the cell apoptosis is inhibited. MMP-16 may be considered as a target gene for therapy of ESCC.

To report a case of fructose-1, 6-bisphosphatase deficiency child with delayed diagnosis for seven years, and review the literature. A 13-year-old boy presented with recurrent episodes of hypoglycemia after infection since 6 years old, he also had convulsion and short stature. The laboratory finding revealed ketotic hypoglycemia, lactic acidosis, transient elevated ALT and UA. The genetic analysis showed compound heterozygous mutations of c. 960-961insG and c. 355G> A in FBP1 gene. Among eighteen patients reported in China, 88.9% had convulsion, 16.7% had growth retardation, the average delayed diagnosis spent 2.8 years, which could result in permanent brain damage and high death risk. The most common mutation was c. 960-961insG, followed by c. 355G>A and c. 490G>A, these mutations maybe hot spot sites of FBPl gene in China.

Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.

Objective To investigate the therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis (SAP) in rats.Methods A total of 80 healthy Sprague-Dawley rats were divided into eight groups:SAP group (three hours、48 hours),Saussurea involucrate treated group (three hours、48 hours),ulinastatin control group (three hours、48 hours) and sham operation group (three hours、48 hours),10 rats in each group.After modeling,the rats of SAP group were regularly feeded and the rats of other three group were treated with Saussurea involucrate injection (1.04 mL/kg) intraperitoneal injection,ulinastatin 10 000 U/L tail vein injection,and saline femoral vein injection,respectively and injected every 12 hours.At three hours and 48 hours after treated,blood and pancreatic tissue samples were obtained.The mortality rate,serum amylase level and pathological changes of the pancreas of each group were observed.Serum tumor necrosis factor alpha (TNF-α),interleukin (IL)-6 and IL-10 levels were detected by enzyme linked immunosorbent assay (ELISA).The content of malondialdehyde (MDA) in pancreatic tissues was determined by chemical colorimetry.The level of TNF-α mRNA,IL 6 mRNA and IL-10 mRNA in pancreatic tissues were measured with reverse trascription-polymerase chain reaction (RT-PCR).The activity of nuclear factor kappa B p65 (NF-κB p65) in the pancreatic tissue was tested by immunohistochemistry.Single factor analysis of variance was used to compare multiple groups,and the least significant difference (LSD) method was used in the multiple comparisons between groups.Fisher's exact probability method was performed for rates comparison.Results At 48 hours,there was no statistically significant difference in mortality rate among Saussurea involucrate treated group,SAP group and ulinastatin groups (all P＞0.05).At 48 hours,the histopathology score (8.13 ± 0.64),levels of serum amylase ((2 597.0±214.0) U/L),TNF-α ((254.4±11.6) ng/L),IL-6 ((441.4±14.6) ng/L),levels of pancreatic tissues MDA ((311.0±10.6) mmol/L),TNF-α mRNA(2.04±0.08),IL-6 mRNA (1.77±0.04)and activity of NF-κB p65 ((25.90±2.90)％) of Saussurea involucrate treated group were all lower than those of SAP group (11.40±0.89,(4 780.0±101.0) U/L,(396.0±7.4) ng/L,(664.4± 7.6) ng/L,(418.0± 10.6) mmol/L,2.94±0.03,2.63±0.08 and (51.60±5.27) ％;however level of serum IL-10 ((133.5±6.9) ng/L vs (95.1±5.2) ng/L) and IL-10 mRNA of the pancreatic tissue (1.38±0.06 vs 0.85±0.03) significantly increased (F=253.07、441.63、489.40、2 465.00、196.65、477.89、562.79、131.70、560.18、570.04,all P＜0.01).There was no significant differences in all above parameters between Saussurea involucrate treated group and ulinastatin groups (7.56±0.88,(2 607.0±239.0) U/L,(252.2 ±9.2) ng/L,(443.4±9.6) ng/L,(308.4±9.2) mmol/L,2.10±0.12,1.74±0.04,(26.00±3.67)％,(134.5±7.8) ng/L and 1.42±0.06) at 48 hours (all P＞0.05).Conclusion Saussurea involucrate injection can eliminate oxygen free radicals and prevent to xidation,inhibit NF-κB activation,regulate synthesis and release of cytokines,and alleviate pancreatic injury in SAP rats,but it can not decrease mortality.

The laboratories of National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, have developed a series of experimental method . These method have unique advantages over the national standard method and industry standard method . How to make these method become public products through legal procedures to serve disease prevention and control more extensively is an obligatory task for national virological laboratories. This article explores the establishment, verification, validation, and examination of virological non-standard method under laboratory certification and accreditation conditions through empirical research on the " RT-RAA method for detecting EV71/CA16 and other viruses" .

Objective:To investigate the level of beliefs about medication in young patients with newly treated pulmonary tuberculosis and analyze its influencing factors.Methods:This was a cross-sectional study. Using the convenient sampling method, a total of 320 young newly treated pulmonary tuberculosis patients who visited the designated tuberculosis hospitals in Kashgar Prefecture and Hotan Prefecture of Xinjiang Uygur Autonomous Region from January 2022 to April 2023 were selected as the research objects. The investigation was carried out with the General Information Questionnaire, Beliefs about Medicines Questionaire Specific (BMQ-S), 8-Item Morisky Medication Adherence Scale (MMAS-8), Brief Illness Perception Questionnaire (BIPQ) and Tuberculosis-related Stigma Scale (TSS). Multiple linear regression was used to analyze the influencing factors of medication beliefs in young patients with newly treated tuberculosis.Results:A total of 320 questionnaires were distributed in this study, and 302 valid questionnaires were collected, with an effective response rate of 94.38% (302/320). The total score of BMQ-S of 302 young patients with newly treated tuberculosis was -1.00 (-2.00, 1.00), MMAS-8 score was 5.38 (4.75, 5.75), BIPQ score was 37.00 (24.00, 44.00) and TSS score was 48.00 (44.00, 52.00). The results of multiple linear regression analysis showed that comorbidities, medication adherence, disease perception and stigma were the influencing factors of medication beliefs in young newly treated pulmonary tuberculosis patients ( P<0.01) . Conclusions:The medication belief level of young patients with newly treated pulmonary tuberculosis needs to be improved. Medical staff should correct the negative cognition of the patient's disease, emphasize the benefits of drug treatment and enhance the patients' beliefs in the necessity of medication.

An integrated segmentation method for 3D ultrasound carotid artery was proposed. 3D ultrasound image was sliced into transverse, coronal and sagittal 2D images on the carotid bifurcation point. Then, the three images were processed respectively, and the carotid artery contours and thickness were obtained finally. This paper tries to overcome the disadvantages of current computer aided diagnosis method, such as high computational complexity, easily introduced subjective errors et al. The proposed method could get the carotid artery overall information rapidly, accurately and completely. It could be transplanted into clinical usage for atherosclerosis diagnosis and prevention.
Algorithms ; Angiography ; methods ; Carotid Arteries ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; methods ; Ultrasonography

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Objective: To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression. Methods: UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by ³²P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager. Results: UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA. Conclusions UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

Objective: To clarify the potential pathogen for fever of unknown origin (FUO) in serum samples for which pathogenic agents were hardly identified with conventional exainatins. Methods: Random capturing the nucleic acid of pathogen was performed by utilizing the property of sequence non-dependence of next generation sequencing (NGS), followed by enrichment of nucleic acid with multiple displacement amplification (MDA). After sequencing, metagenomic analysis was applied to the raw data and the phylogenetic tree was built to identify the potential pathogen. Results: The result did not indicate common pathogens for FUO but showed the existence of Torque teno Viurs (TTV). Assembly was carried out to all sequencing reads. The coverage of consensus sequence on reference was calculated. Phylogenetic result indicated that all confidence sequences belonged to 3 genera (α TTV, β TTV and γ TTV). Conclusions The characteristics of genome, phylogenesis of TTV and TTV as signal at immunology level were analyzed and clarified. The possible explanation for detection of TTV in 3 genera may be that TTV itself or non-infectious factors caused the immunosuppression, which finally result e in rise of TTV detection.