OBJECTIVE:To provide reference indexs for the quality evaluation of Glycyrrhetinic acid lipo-emul. METHODS:Croy-TEM was used to detect the morphology of Glycyrrhetinic acid lipo-emul,laser nano-particle size analyzer was used to deter-mine the particle size,polydispersity index(PDI)and the zeta potential;UPLC was used to determine the drug loading of its ac-tive ingredient glycyrrhetinic acid;placing 10 d in 30 ℃,then stability was detected. RESULTS:Prepared Glycyrrhetinic acid li-po-emul was clear outline,structural integrity,roundlike and arranged closely;the mean particle size was(245.2±4.29)nm,PDI was (0.054 ± 0.01) and the mean zeta potential was (-6.25 ± 0.54) mV;the average drug loading of glycyrrhetinic acid was (1.25±0.09)mg/mL;the sample with mass concentration of 0.82 mg/mL showed good stability in within 10 d. CONCLUSIONS:Particle size,zeta potential,drug loading and stability can be used as the evaluation index of Glycyrrhetinic acid lipo-emul.

OBJECTIVE:To study the effect of hydrophilic/hydrophobic nano-silica with different adding amount on the stabili-ty of lipo-emulsion. METHODS:Glycyrrhetinic acid lipo-emulsion 4 mL was taken,respectively adding into 0.5%,1.0%,1.5%(m/m,the same below) hydrophilic nSiO2,and 0.4%,0.75,1.0% hydrophobic nSiO2,incubating 2 h in 30 ℃ water;the same batch of Glycyrrhetinic acid lipo-emulsion was treated as blank control. The forms were observed under electron microscopy after treatment,absorbance value was determined,the stability parameter (KE) was calculated according to the absorbance value,then the adding amount of nSiO2 was optimized,3 batches of preparations was prepared,and the verification test was conducted. RE-SULTS:The spherical structure was Glycyrrhetinic acid lipo-emulsion in the electron microscopy,the substance wrapping its sur-face white ring (fully wrapped) or semi-circular structure (not fully wrapped) was nSiO2. KE of hydrophilic nSiO2 were 4.66%, 5.01% and -2.08%,and KE of hydrophobic nSiO2 were 3.02%,4.51% and 7.24%. The optimized adding amount of hydrophilic nSiO2 was 0.2%,0.3% and 0.4%,hydrophobic nSiO2 was 0.1%,0.2% and 0.3%;KE were 6.19%,3.05%,7.84%,8.42%, 2.41%,2.93%,respectively. The optimal adding amount was 0.3% hydrophilic nSiO2 and 0.2% hydrophobic nSiO2;the 3 batches of preparation showed the optimum stability in its own adding amount. CONCLUSIONS:Both hydrophilic and hydrophobic nSiO2 can improve the stability of Glycyrrhetinic acid lipo-emulsion,and preferably 0.3%,0.2%.

Objective To investigate the effects of alcohol on hepatitis C virus( HCV) replication and type I interferon signaling pathway in human hepatocytes .Methods Primary hepatocytes were treated with different concentrations of alcohol , and then infected with HCV .The infected cells were collected to measure the level of HCV RNA .The alcohol-treated hepatocytes were also collected to detect the expression of HCV Core, IFN-α, IFN-β, IRF-7, suppressor of cytokine signaling SOCS-2 and SOCS-3 at mRNA and protein levels by real-time quantitative PCR and ELISA or Western blot , respectively .Results Alcohol treatment enhanced HCV infection and replication in primary hepatocytes at concentrations higher than 10 mmol/L in a dose-dependent manner (P<0.05).Treatment with 40 mmol/L of alcohol significantly reduced the expression of IFN-α, IFN-βand IRF-7 at mRNA and protein levels , and increased the expression of SOCS-2 and SOCS-3 at mRNA and protein levels .Conclusion Alcohol treatment could damage the host in-nate immunity in human hepatocytes and promote HCV replication by reducing the expression of type Ⅰinter-feron ( IFN-αand IFN-β) and IRF-7 and increasing the expression of negative regulators including SOCS-2 and SOCS-3.These results demonstrated that the impairment of innate immunity in liver of alcohol abusers might contribute to the enhancement of HCV infection and result in poor therapeutic effect of IFN -α.

Objective To study the phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus (HIV )‐1 strains in Guangxi Zhuang Autonomous Region . Methods Plasma samples of 158 HIV‐1 infected patients in Guangxi area were collected during October 2011 to March 2012 .The gag gene fragments of HIV‐1 were amplified by reverse transcription/nested‐polymerase chain reaction and then sequenced .MEGA 5 .03 was utilized to construct phylogenetic tree and to calculate the genetic distances and selection pressures (globle ω) of gag gene and its coding regions . The comparisons between two groups were tested by Student′s t test ,and the comparisons of multiple groups were tested by one‐way ANOVA .Results A total of 140 amplification products of gag gene were obtained from 158 samples .Four subtypes of HIV‐1 were found ,including CRF01_AE (80 ,57 .1% ) , CRF08_BC (46 ,32 .9% ) ,CRF07_BC (10 ,7 .1% ) ,and subtype B (B′) (4 ,2 .9% ) .The genetic distances of gag gene of the above subtypes were 0 .036 ± 0 .001 ,0 .031 ± 0 .002 ,0 .043 ± 0 .003 and 0 .102 ± 0 .006 ,respectively ,with statistical significance (F=220 .62 ,P<0 .01) .The p17 and p24 coding regions suffered negative selection pressure (globleω<1) .Neither the globle ω in p17 region nor that in p24 region had significant differences among different subtypes (F=0 .761 ,P=0 .469 and F=0 .037 ,P=0 .964 , respectively ) . Conclusion CRF01_AE is the major subtypes of HIV‐1 in Guangxi Zhuang Autonomous Region .The coding regions of gag gene are relatively conserved during evolution .Changes of HIV‐1 prevalence ,however ,may affect the genetic variation of gag gene ,which should be continuously monitored .

Abstract: Upon monitoring cytoplasmic aberrant double-stranded DNA, cGAS-STING signaling pathway induces the expression of type I interferons and pro-inflammatory cytokines, which activates the host immune response and enhances anti-tumor immune response and resistance to pathogen infection. However, sustained activation of the cGAS-STING signaling pathway drives diseases such as autoimmune diseases, aging-associated inflammation, and neurodegenerative pathologies. Herein, we describe the mechanism by which cGAS-STING signaling pathway participates in regulating the development of various immune-related diseases, with a particular review of the research and development progress of STING agonists, cGAS inhibitors, and STING inhibitors, aiming to provide some theoretical reference for the future development of cGAS-STING modulators.

Objective:To investigate the method of determining oral implantation sites based on an anatomical model of mandibular premolar area of a Beagle dog.Methods:This study was performed in the Second Affiliated Hospital of Zhejiang Chinese Medical University between January 2019 and October 2020. Mandibular anatomical structure and measurement data were compared between before and after removal of premolar teeth to determine safe implantation areas and oral implantation sites.Results:Among all mandibular premolars, the roots of the 1 st to 4 th premolars (P1-P4) gradually increased. The diameter of the mesial roots of the double root teeth P2, P3, and P4 was (2.72 ± 0.45) mm, (3.22 ± 0.32) mm, (4.16 ± 0.34) mm, respectively, which was significantly shorter than those in the distal roots [P2: (2.98 ± 0.29) mm, P3: (3.48 ± 0.27) mm, P4: (4.58 ± 0.22) mm]. The length of distal roots P2, P3 and P4 was (8.79 ± 0.41) mm, (9.21 ± 0.31) mm, (10.12 ± 0.36) mm), respectively, which was significantly shorter than that of mesial root [P2: (8.91 ± 0.69) mm, P3: (9.48 ± 0.27) mm, P4: (11.58 ± 0.24) mm]. Among all mandibles, the distance (H) from the mental foramen to the first molar and the width (W) of the alveolar crest increased successively [H1: (7.24 ± 0.49) mm, H2: (8.28 ± 0.71) mm, H3: (9.52 ± 0.37) mm, W1: (5.71 ± 0.81) mm, W2: (5.82 ± 0.28) mm, W3: (6.72 ± 0.54) mm]. Conclusion:The mental foramen and the distal part outside the canine apical area are safe implantation areas. In the safe implantation area, the length and diameter of the implant prosthesis do not exceed the root length in the implantation area and the maximum diameter in the buccal lingual direction.

Objective To investigate the HIV-1 drug resistance in Guangxi during 2009 to 2012 and to analyze the correlations between drug resistance and HIV-1 subtypes.Methods Patients with human immunodeficiency virus infection or acquired immune deficiency syndrome ( HIV/AIDS) were randomly re-cruited from different areas in Guangxi.HIV-1 RNA was extracted from blood samples of the subjects and converted into complementary DNA ( cDNA) by using reverse transcription.The pol gene was amplified and sequenced.Subtyping analysis was performed by using the online analysis tool of Genotyping in combination with the MEGA 5.03 software.The HIV resistance mutations were determined and scored with the use of Stanford HIV Drug Resistance Database.Results A total of 196 pol gene sequences were obtained from 103 antiretroviral therapy (ART)-treated subjects (52.55%) and 93 ART-na?ve subjects (47.45%).The 196 pol gene sequences were classified into four subtypes including CRF01_AE, CRF08_BC, CRF07_BC and B, accounting for 48.47%, 44.90%, 6.12%and 0.51%, respectively.The HIV drug resistance rates in sub-jects with and without ART were 10.68% and 7.53%, respectively.Among the 196 subjects, 14 cases showed low level of drug resistance, 3 cases showed moderate level of drug resistance and 4 cases showed high level of resistance.Only one case was resistant to both nucleoside reverse transcriptase inhibitors ( NR-TIs) and non-nucleoside reverse transcriptase inhibitors ( NNRTIs) .The resistance rates of the 196 cases to protease inhibitor (PIs), NRTIs, NNRTIs, and integrase inhibitors (INs) were 6.63%, 3.06%, 11.22%and 8.67%, respectively.The frequencies of PIs-related mutations in subtypes CRF01_AE, CRF07_BC and CRF08_BC were 6.32%, 41.67% and 2.27%, respectively.Most of the PI-related A71V/T mutations were identified in strains belonging to subtype CRF07_BC, accounting for 75% of all A71V/T mutations found in the 196 strains.The NNRTI-related E138A mutations only appeared in strains belonging to subtype CRF08_BC.Conclusion The drug resistance rate among patients with HIV-1/AIDS in Guangxi was higher than the average level in China.The drug resistance rates varied with the subtypes of HIV-1 strains.

OBJECTIVETo investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.

METHODS152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.

RESULTSAmong 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.

CONCLUSIONCRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.

Blotting, Western ; China ; epidemiology ; Cities ; Drug Users ; Genes, pol ; Genotype ; HIV Infections ; epidemiology ; transmission ; virology ; HIV-1 ; genetics ; Humans ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; blood ; pol Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo investigate the impact of heroin for antiviral treatment, drug resistance, mutation types and frequency in HIV/AIDS patients in Guangxi Zhuang Autonomous Region.

METHODSHIV/AIDS patients were recruited in Methadone Maintenance Treatment Clinics, HIV/AIDS Clinic and HIV Voluntary Counseling and Testing Center Liuzhou and Baise city from April 2008 to October 2009. The patients were grouped by the situation of antiviral treatment and use of heroin. A total of 435 HIV/AIDS patients were recruited, among which 108 cases in antiviral treatment and heroin group, 93 cases in antiviral treatment and never using drug group, 105 cases in no antiviral treatment and using heroin group, 129 cases in no antiviral treatment and never using drug group. The effect of antiviral treatment was evaluated by questionnaire survey, viral load measurement and CD4(+) T lymphocyte count. HIV-1 RNA from plasma was extracted, and then the pol genes were amplified and sequenced. The sequences were analyzed for HIV-1 genotype drug-resistance.

RESULTSFor the patients who received antiviral treatment, the viral load in heroin group was higher than that in never using drug group (lg (2.61 ± 1.24) vs lg (2.08 ± 0.80), t = 3.54, P < 0.05) , and the percentage of viral load lower than 1 000 copies/ml in heroin group was significantly less than that in never using drug group (63.9% vs 86.0%,χ(2) = 12.76, P < 0.05). For the patients who received antiviral treatment, the difference has no significance in CD4(+) T lymphocyte count between heroin group and never using drug group ((337.92 ± 181.66) vs (326.14 ± 254.98), t = 0.38, P = 0.703). For the patients who didn't receive antiviral treatment, the difference also has no significance in CD4(+) T lymphocyte count between heroin group and never using drug group ((373.73 ± 155.97) vs (337.53 ± 209.26), t = 1.47, P = 0.143). For the patients who received antiviral treatment, there was no difference in the percentage of the CD4(+) T lymphocyte count more than 350/ml between heroin group and never using drug group (48.1% vs 43.0%, χ(2) = 0.53, P = 0.466). 319 HIV-1 pol gene sequences were obtained. Among the patients who received antiviral treatment, the mutation frequency of M184V/I, T215Y/F, L210W and T69N/S in heroin abuser group were significantly higher than that in never using drug group (14.9% (11/74) vs 4.4% (3/68), 12.2% (9/74) vs 1.5% (1/68), 12.2% (9/74) vs 1.5% (1/68) and 10.8% (8/74) vs 1.5% (1/68) respectively) (P < 0.05).

CONCLUSIONUsing heroin may promote HIV replication, reducing the virological response to antiviral treatment and increasing the frequencies of drug resistance loci among HIV/AIDS patients.Heroin rehabilitation may benefit from the antiviral treatment and obtain better antiviral effect.

Acquired Immunodeficiency Syndrome ; Anti-HIV Agents ; Antiviral Agents ; CD4 Lymphocyte Count ; China ; Drug Resistance ; Drug Resistance, Viral ; Genes, pol ; HIV Infections ; HIV-1 ; Heroin ; adverse effects ; Heroin Dependence ; Humans ; Mutation ; drug effects ; Mutation Rate ; Viral Load