Currently,there are a lot of targets for the treatment of HBV infection,both for the host,and for the virus itself.However,existed clinical drugs can only control HBV infection,and can not remove the HBV,especially cccDNA.Therefore,the chronic persistent infection caused by HBV related diseases is still seriously threat to human health.People are still impatient for the development of new effective anti-HBV drugs.In this paper,we review the recent research of the anti-hepatitis B virus based on the host as the target.

Aim To investigate the method of cell culture for smooth muscle cells from rat cerebral basilar artery and understand cells growth and biological characteristics.Methods The explant attached method was applied for cell culture of rat basilar artery smooth muscle cells(BASMCs).The cultured BASMCs were identified by immunocytochemical staining.The activities of cells were indicated by the dynamic changes of intracellular calcium concentration observed by RF-5 000 fluorospectro-photometer.Results BASMCs grew out of tissue blocks by 5 days,reached confluency could be subcultured after 2 weeks.Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Introduction of calcium channel agonists induced significant increase in Fura-2 fluorescence ratio(F340/F380)and cells were in good condiction.Conclusion Explant attached method is simple,efficient and economic.It provides an ideal cell model for the study of pathogenesis of the cerebral vascular diseases.

Objective Using shotgun mass spectrometry to detect proteins probably contained in bone marrow stromal cells(BMSCs) conditioned medium. MethodsMixed with BMSCs conditioned medium was divided into two parts which is(>5kD and <5kD) by means of ultrafiltration. The two parts were used to culture neural stem cells(NSCs) separately, and the proportions of neurons, astrocytes and oligodendrocytes in the offsprings of NSCs were calculated, then the effective part that could regulate the differentiation of NSCs was detected by Shotgun mass spectrometry. Results The BMSCs conditioned medium which is >5kD could promote the NSCs differentiate into more neurons and oligodendrocytes. The SDS-PAGE of this part showed that the most proteins were above 14kD, then the protein bands were enzymed. In total, 456 proteins were identified by Shotgun mass spectrometry after all the protein bands were enzymed, there were 154 similar proteins, 17 hypothetical proteins and 56 unknown proteins. And in the rest of 229 proteins, most of them were cytoskeletal proteins, secreted proteins, signal transduction proteins, enzymes, transporter and so on. Conclusion Many proteins secreted by BMSCs could regulate the differentiation of NSCs so as to prove the protein components probably existed in the BMSCs conditioned medium.

In the present study, the total RNA was extracted from three instar larvae of Musca domestica, the cDNA sequence encoding the ORF of cecropins was amplified by RT-PCR, and the target fragment was further sequenced after being cloned into T vector pUCm-T. Then, the cDNA sequence of the mature cecropins was amplified by PCR with recombinant plasmid pUCm-T/cecropin as template, the N-terminal rare codon GGA of E. coli was changed to the favorable codon GGC,and a Asn codon AAC was added in front of the stop coden TAA in the C- terminus. This mutant gene designated as mCecropin was then ligated with the fusion expression vector pGEX-4T-1. After restriction analysis and DNA sequencing, the positive recombinant plasmid pGEX-4T-1/mCecropin was transformed to different strains of E. coli cells and the fusion protein was expressed after IPTG induction. The fusion protein was assayed by SDS-PAGE and the E. coli BL21(DE3) cell was chosen as the host cell for the expression of the fusion protein. The expressed fusion protein GST-mCecropin was purified by GSTrap affinity coloum and the GST marker was then cleaved by thrombin. In this way, the fusion protein mCecropin with antibacterial activity was obtained after purification with HiTrap benzamidine column.

Objective To investigate the diagnostic value of matrix gamma carboxy glutamic acid protein(MGP) for coronary heart disease(CHD) .Methods Enzyme linked immunosorbent assay was performed for the detection of serum MGP level in health‐y subjects and CHD patients with different coronary artery calcium score(CACS) .Receiver operating characteristic(ROC) curve was used to analyze the diagnostic value of MGP for CHD .Results Between CHD patients and healthy subjects ,and CHD patients with different CACS ,the difference of serum MGP level was significant ,and serum MGP level was positively correlated with CACS (P<0 .05) .ROC curve of showed that the area under ROC curve was 0 .667 ,the diagnostic threshold was 70 .69 pg/mL ,the diag‐nostic sensitivity was 58 .80% ,the specificity was 83 .70% and the Youden index was 0 .425 .Conclusion CHD patients might be with abnormal serum MGP level ,which could be positively correlated with CACS .MGP might be with significant value for the diag‐nosis of CHD ,could be useful for the clinical prevention and early diagnosis of CHD .

China is a country with a high prevalence of myopia,and the incidence of myopia among adolescents is in-creasing year by year.The usage rate of orthokeratology is also increasing year by year.In order to improve the effectiveness of myopia control,ophthalmologists have adopted plans such as optimizing lens design and combined treatment to provide protection for adolescent myopia control.This article reviews the diverse clinical applications of orthokeratology and their effectiveness based on national and international literature.

OBJECTIVETo construct a lentiviral vector carrying the γ-synuclein(SNCG) gene and establish a human colorectal carcinoma cell line SW1116 stably expressing this gene, and then investigate the inhibition of the growth and invasion capacity of SW1116 cells.

METHODSRNA interference fragment was designed according to the SNCG sequence (GenBank: No.NM003087.2), and then SNCG RNAi effective target genes were screened. After the Oligo DNA of target sequences was synthesized, the lentiviral vectors carrying LV-SNCG-RNAi-EGFP (RNAi group) and LV-SNCG-NC-EGFP (NC group) were constructed and packaged to produce lentivirus venom. The supernatants of different virus-producing cells were used to transfect SW1116 cells respectively. Wild SW1116 cells were used as blank control (CON group) EGFP fluorescence was detected by fluorescent microscopy and the differential expression of SNCG mRNA and protein was detected by real-time PCR and Western blot. CCK-8, soft agar assay and Transwell chamber were employed to estimate the inhibiting effect on growth and invasion of SW1116 respectively.

RESULTSRecombinant lentiviral vectors respectively carrying the SNCG-RNAi-EGFP and SNCG-NC-EGFP were successfully constructed and the supernatants of lentivirus could effectively infect SW1116 cells. The titer of the virus carrying LV-SNCG-RNAi-EGFP or LV-SNCG-NC-EGFP was 8×10(8) TU/ml. Real-time PCR and Western blot confirmed that compared with the NC group, SNCG-RNAi group had lower SNCG expression (1.009±0.161 vs. 0.114±0.030, P=0.009), and showed tremendous silencing effect as 76.8%(P<0.05). SNCG protein expression was also significantly reduced (RNAi:12.001±2.884, NC:32.443±4.731, CON:34.308±6.920, P<0.05). After SNCG knockdown, the number of proliferation cells was obviously reduced at 48, 72, 96 and 120 hours respectively(P=0.036). In soft agar assay, clones in RNAi group were smaller[RNAi:(0.582±0.103) mm, NC:(1.863±0.316) mm, CON:(1.749±0.525) mm]. Colony formation rate of RNAi group was down to (17.1±3.5)%, which was significantly lower than (36.5±4.3)% in NC group and (33.8±3.9)% in CON group. In migration test, the number of invasion cell was 37.4±9.3 in RNAi group, which was significantly less than 112.3±8.6 in NC group and 100±0.0 in CON group.

CONCLUSIONExpression of SNCG mRNA and protein plays an important role in the growth and the invasion capacity of SW1116 cells.

Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; Humans ; Lentivirus ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Transfection ; gamma-Synuclein ; genetics

Objective:To understand the impact of intimate social networks and sexual networks on HIV-infection in men aged ≥50 years in Pengzhou, Sichuan Province, and provide reference for the formulation of HIV/AIDS prevention and control strategies in middle-aged and elderly people.Methods:A case-control study was conducted by an interview-style questionnaire survey in a case group consisted of 114 men aged ≥50 years and a control group consisted of 423 healthy men matched by age frequency in Pengzhou, Sichuan Province, from April 2019 to October 2020, and the influencing factors of HIV infection were identified by unconditional stepwise logistic regression analysis.Results:Among the 114 cases, 63.16% (72/114) and 96.49% (110/114) of men had intimate social network size and sexual network size ≥4 people, which were higher than the control group of 23.40% (99/423) and 11.58% (49/423) (all P<0.05).The overlap of intimate social network and sexual network was 49.12% (56/114) in the case group, which was lower than that in the control group (82.27%, 348/423)( P<0.05). Logistic regression analysis showed that the size of the intimate social network ≥4 people ( OR=8.66, 95 %CI: 2.01-37.30), the size of the sexual network ≥4 people ( OR=1 121.11, 95% CI: 28.38-4 429.61), the material support from the first intimate network member ( OR=6.39, 95% CI: 1.03-39.75), and multiple sexual partners ( OR=55.50, 95% CI: 6.22-494.96) were the risk factors of HIV-infection in men aged ≥50 years. Receiving health education about AIDS ( OR=0.07, 95% CI: 0.01-0.37), high sexual satisfaction with the first sexual partner and good personal relationship ( OR=0.02, 95% CI: 0.01-0.59; OR=0.01, 95% CI: 0.01-0.26), and the overlap between intimate social network and sexual network ( OR=0.02, 95% CI: 0.01-0.38) were the protective factors of HIV-infection in men aged ≥50 years. Conclusions:The overlap of intimate social network and sexual network can reduce the risk of HIV-infection in men aged ≥50 years. It is important to help middle-aged and elderly men to build good and stable intimate social and sexual networks to meet their social communication and sex demands and prevent HIV infection by reducing high riak sexual behaviors.

Objective:To explore clinical features, diagnosis, localization, and therapeutic strategy of migratory pharyngeal and cervical esophageal foreign bodies.Methods:A total 23 cases of pharyngeal and cervical esophageal migratory foreign bodies were admitted between January 2015 and December 2021. There were 14 females and 9 males with the age ranged from 35 to 82 (55.0±12.7)years. In all the cases, esophageal CT was taken to confirm the esophageal foreign body. Multiplanar reconstruction (MPR) was performed to locate the foreign body from the horizontal, coronal and sagittal dimensions as well as the corrected reconstructed MPR. According to the location of the foreign body, appropriate surgical method was selected.The symptoms, complications, types of foreign body, positioning, surgical methods, and relevant information were recorded.Data were analyzed using the descriptive method and SPSS 25.0 software.Results:The clinical symptoms of 23 migrating esophageal foreign bodies included pharyngodynia (20/23), foreign body sensation (6/23), hoarsenss (1/23), difficulty in turning neck(1/23), difficulty in opening mouth (1/23), fever (7/23), poor appetite (1/23), and abdominal pain (1/23). The foreign bodies included 19 fish bones, 2 wires, 1 embroidery needle and 1 chicken bone. There were 9 cases (39.1%) of foreign bodies located in extraluminal cervical esophagus, 2 cases (8.7%) of foreign bodies located in the muscular layer of the cervical esophagus and 12 cases (52.2%) of foreign bodies located in pharynx. Twenty-one cases of foreign bodies were removed by cervical lateral incision, in which 11 were removed by cervical lateral incision directly, 10 by the second lateral cervical incision after the foreign bodies were accurately located by MPR and/or corrected MPR, 1 foreign body was removed by incision of the pharyngeal mucosa under suspension laryngoscope, 1 foreign body was removed by tracheoscopy. Compared with patients with intraluminal foreign bodies ( n=308) treated in the same period, intake of fishbone [19 (19/23) vs. 133 (82.6% (43.2%, 133/308), OR=7.31] and first visit was more than 24 hours [20(87.0%, 20/23) vs. 77(25.0%, 77/308), OR=17.2] were the significant risk factors of migratory esophageal foreign bodies. Conclusions:MPR and the corrected MPR can accurately locate the migrating pharyngeal and cervical esophageal foreign bodies, by providing more intuitive imaging evidence for doctors, which provide imaging basis for formulation of surgical programs. Foreign bodies in pharyngeal and cervical esophagus need to be treated as soon as possible, otherwise they are easy to migrate, leading lead to serious complications.

Hepatic angiomyolipoma （HAML） is a rare benign mesenchymal tumor frequently observed in middle-aged women. Due to the absence of prominent symptoms in the early stage and the lack of specific imaging findings， the diagnosis of this disease can be challenging， leading to a high rate of misdiagnosis. This article reports a case of giant HAML with subcapsular rupture that was misdiagnosed as hepatocellular carcinoma and introduces the characteristics of the case and its diagnosis and treatment process， in order to provide a reference for the diagnosis and treatment of this type of disease.