In vitro antifungal activity of Shikonin against Candida albicans by inducing cellular apoptosis and necrosis.

BACKGROUND: Our previous studies showed that Shikonin (SK) had a strong anti-Candida albican (C. albicans) activity, especially against some fluconazole-resistant strains, which is probably due to the oxidative damage of SK to C. albicans. METHODS AND RESULTS: In this study, we expanded the antifungal spectrum and evaluate the toxicity of SK. The results indicated that SK also exhibited potent invitro antifungal activities against other pathogenic fungi such as other Candida, Aspergillus, Cryptococcus, and Dermatophytes, but did not display apparent toxicity to the mammalian cells, suggesting that SK is safe to be a potential antifungal drug. Furtherly, we analyze the exact mechanism of SK against C. albicans. We found that SK could induce a series of apoptosis characteristics, including phosphatidylserine externalization, chromatin condensation and fragmentation, decreased cytochrome c oxidase activity as well as caspase activation. CONCLUSIONS: In summary, this study highlighted the antifungal activity and mechanism of SK against C. albicans, providing a potential therapeutic strategy for C. albicans infection.

Men who have sex with men is the high-risk drug resistance population: A meta-analysis of HIV-1 drug resistance profiles and trends in China.

WHAT IS KNOWN AND OBJECTIVE: Except for the transmitted drug resistance (TDR)'s standard sampling and monitoring protocol, China's HIV-1 pretreatment drug resistance (PDR) and acquired drug resistance (ADR) results vary widely due to the studies' diverse background. This meta-analysis was conducted to comprehensively understand the drug resistance profiles of Chinese HIV/AIDS patients and compare the drug resistance differences among groups to provide a reference for the further improvement of treatment protocols. METHODS: Data sources for this study were WANFANG, VIP, CNKI, SinoMed, PubMed and Web of Science databases from January 1, 2010 to January 13, 2022. Data extracted from the literature were analyzed by R and Stata to evaluate the profile and changing trend of drug resistance in Chinese HIV/AIDS patients. RESULTS AND DISCUSSION: One hundred twenty-one literature were included. The combined PDR and ADR in the Chinese HIV/AIDS patients was 5.56% (95% CI: 4.77%-6.41%) and 51.33% (95% CI: 47.57%-55.38%), respectively. The time trend analysis shows the upward trend of PDR. There were significate differences in ADR among different sample sources, the ADR in the central region were higher than those in all other regions. The PDR in men who have sex with men (MSM) was lower the whole population, while the MSM's ADR was much higher than whole population. WHAT IS NEW AND CONCLUSION: PDR in China showed an upward trend and exceeded the 5% warning line but is still at a low level worldwide. ADR is stable and below the middle level globally; the comprehensively promoted free ART in China still has lasting effects. MSM is the high-risk drug resistance population, targeted treatment strategies should be used.

First Detection of a Cluster Novel HIV-1 Second-Generation Recombinant (CRF01_AE/CRF07_BC) among Men Who Have Sex with Men in Nanjing, Eastern China.

INTRODUCTION: A large number of unique recombinant forms have been found in China in recent years. This study aimed to report on a cluster of novel HIV-1 recombinants. METHODS: We constructed phylogenetic trees using the maximum likelihood (ML) method with 1,000 bootstrap replicates in IQ-TREE 1.6.8 software and determined recombination break points using SimPlot 3.5.1. RESULTS: Overall, 9 near-full-length genome (NFLG) sequences were reported in this study, including 1 circulation recombinant form (CRF)01_AE NFLG sequence and 8 highly similar novel HIV-1 second-generation recombinants composed of CRF01_AE and CRF07_BC (CRF105_0107) isolated from a cluster HIV-positive male subjects infected among men who have sex with men (MSM) in Nanjing, eastern China. The phylogenetic analysis of NFLG showed 1 sequence named "nj16" to have at least 11 breakpoints inner virus and 7 other sequences to have at least 10 breakpoints inner virus. Our findings further showed as follows: first, this is the first time that a cluster of novel CRF105_0107 HIV-1 strains were identified among MSM in Nanjing, Jiangsu. Second, the Chinese "4a" cluster of CRF01_AE which mainly circulating in northern China has spread in Jiangsu for more than 15 years. Third, HIV-1 recombination events were active in Nanjing city, and novel recombinants could spread rapidly through some small-scale transmission networks. CONCLUSION: The continued emergence of novel recombinant HIV-1 strains in Nanjing suggests dynamics and complexity in the HIV epidemic among MSM in Jiangsu province. Further investigations and molecular epidemiological research should be taken to monitor and understand transmission networks among MSM.

Use of capillary glucose combined with other non-laboratory examinations to screen for diabetes and prediabetes.

AIM: To evaluate the value and feasibility of capillary glucose assessment, combined with other non-laboratory measures, in screening for diabetes and prediabetes in the community. METHODS: In this cross-sectional study, we assessed fasting capillary glucose, fasting plasma glucose, and both capillary glucose and plasma glucose values after 2-h oral glucose tolerance tests in a total of 3736 samples. We determined the optimal threshold of capillary glucose using receiver-operating characteristic curve analysis. The effect of screening methods using capillary glucose combined with other variables, such as age, BMI and waist circumference, was assessed according to area under the receiver-operating characteristic curve. RESULTS: There was a strong positive correlation between capillary glucose and venous plasma glucose. The area under the curve for the model using fasting capillary glucose to screen for impaired fasting glucose was 0.722, while that for the model using capillary glucose after a 2-h oral glucose tolerance test to screen for impaired glucose tolerance was 0.916. The area under the curve for the model using fasting capillary glucose to screen for diabetes was 0.835, while that for the model using 2-h oral glucose tolerance test capillary glucose was 0.912. The area under the curve for the model using fasting capillary glucose + 2-h oral glucose tolerance test capillary glucose to screen for diabetes was 0.945. The discriminatory capability of models using capillary glucose was somewhat improved by adding non-laboratory variables. CONCLUSIONS: Capillary glucose could be an alternative for screening for diabetes and prediabetes, especially in low-resource areas.

[Effects of diets with different n-6/n-3 fatty acids on cardiovascular risk factors in mice fed high-fat diets].

UNLABELLED: To investigate effects of diets with different fatty acid composition on serum lipid profiles, inflammation, oxidative stress and endothelial function in mice fed high-fat diets. METHODS: Male KM mice were randomly divided into five groups and were fed normal control diet, high-fat lard diet, high-fat diets with n-6/n-3 polyunsaturated fatty acid (PUFA) ratios of 1:1, 5:1 nd 2:1 for fiv weeks, respectively. The levels of serum triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C ) iterlekin-6 (IL-6), alonildehyde (MDA), hypersensitive C-reactive protein (hs-CRP), tumor ncrosis factor alpha (TNF-alpha), lipid lphaproxide (LPO), 8-iso postaglandin F2aalpha (8-iso-PGlphaF2u), oxidialphaed low-density lipoprotein (ox-LDL), free faty acid (FFA), E-selectin (ES) and von Willebrand factor (vWF) were measured. RESULTS: The levels of serum LDL-C and non-HDL-C in the lard group were significantly higher than those in the other groups (P < 0.05). Th e ard group had. significntly higher serum TG and TC concentrations compared to 1:1 and 5: groups P <0. 05). The evels of serum FFA in 20:1 group wre significantly higher than those in 1:1 and 5:1 groups P < 0.05). Co mpred with the lard and 20:1 groups,the 1:1 and 5:1 groups displayed lower levels of inflammatory cytokines, oxidative stress and ES. The 5:1 group sgnificantly decreased the level of serum vWF compared to lard and 20:1 groups P < 0.05). CONCLUSION: diet with low n-6/n-3 PUFA ratio could improve lipid metabolism, inflammation, oxidative stress and endothelial function compared to high-fat diets with lard and higher n-6/n-3 PUFA ratio. The diet with low n-6/n-3 PUFA ratio can improve cardiovascular disease risk factors to prevent cardiovascular disease.

Shikonin Inhibits Candida albicans Biofilms via the Ras1-cAMP-Efg1 Signalling Pathway.

Objective: To investigate the influence of shikonin (SK) on the formation of Candida albicans biofilms and discuss the possible mechanism. Methods: The inhibition of the formation of C. albicans biofilms by SK was observed by scanning electron microscopy. A silicone film method and a water-hydrocarbon two-phase assay were performed to investigate the effects of SK on cell adhesion. Real-time reverse-transcription polymerase chain reaction was used to analyse the expression of genes related to cell adhesion and Ras1-cyclic adenosine monophosphate (cAMP) - enhanced filamentous growth protein 1 (Efg1) signalling pathway. Finally, the level of cAMP in C. albicans was detected and exogenous cAMP rescue experiment was conducted. Results: The results showed that SK could destroy the typical three-dimensional structure of the biofilms, inhibit cell surface hydrophobicity and cell adhesion, downregulate the expression of Ras1-cAMP-Efg1 signalling pathway-related genes (ECE1, HWP1, ALS3, RAS1, CYR1, EFG1 and TEC1) and effectively reduce the production of key messenger cAMP in the Ras1-cAMP-Efg1 pathway. Meanwhile, exogenous cAMP reversed the inhibitory effect of SK on biofilms formation. Conclusion: Our results suggest that SK exhibits potential anti-C. albicans biofilms effects related to the inhibition of Ras1-cAMP-Efg1 pathway.

Jinhua Qinggan granules attenuates acute lung injury by promotion of neutrophil apoptosis and inhibition of TLR4/MyD88/NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Acute lung injury (ALI) is one of the fatal complications of respiratory virus infections such as influenza virus and coronavirus, which has high clinical morbidity and mortality. Jinhua Qinggan granules (JHQG) has been approved by China Food and Drug Administration in the treatment of H1N1 influenza and mild or moderate novel coronavirus disease 2019 (COVID-19), which is an herbal formula developed based on Maxingshigan decoction and Yinqiao powder that have been used to respiratory diseases in China for thousands of years. However, the underlying mechanism of JHQG in treating infectious diseases remains unclear. AIM OF THE STUDY: This study investigated the effects of JHQG on neutrophil apoptosis and key signaling pathways in lipopolysaccharide (LPS) -induced ALI mice in order to explore its mechanism of anti-inflammation. MATERIALS AND METHODS: The effect of JHQG on survival rate was observed in septic mouse model by intraperitoneal injection of LPS (20 mg/kg). To better pharmacological evaluation, the mice received an intratracheal injection of 5 mg/kg LPS. Lung histopathological changes, wet-to-dry ratio of the lungs, and MPO activity in the lungs and total protein concentration, total cells number, TNF-α, IL-1ß, IL-6, and MIP-2 levels in BALF were assessed. Neutrophil apoptosis rate was detected by Ly6G-APC/Annexin V-FITC staining. Key proteins associated with apoptosis including caspase 3/7 activity, Bcl-xL and Mcl-1 were measured by flow cytometry and confocal microscope, respectively. TLR4 receptor and its downstream signaling were analyzed by Western blot assay and immunofluorescence, respectively. RESULTS: JHQG treatment at either 6 or 12 g/kg/day resulted in 20% increase of survival in 20 mg/kg LPS-induced mice. In the model of 5 mg/kg LPS-induced mice, JHQG obviously decreased the total protein concentration in BALF, wet-to-dry ratio of the lungs, and lung histological damage. It also attenuated the MPO activity and the proportion of Ly6G staining positive neutrophils in the lungs, as well as the MIP-2 levels in BALF were reduced. JHQG inhibited the expression of Mcl-1 and Bcl-xL and enhanced caspase-3/7 activity, indicating that JHQG partially acted in promoting neutrophil apoptosis via intrinsic mitochondrial apoptotic pathway. The levels of TNF-α, IL-1ß, and IL-6 were significantly declined in LPS-induced mice treated with JHQG. Furthermore, JHQG reduced the protein expression of TLR4, MyD88, p-p65 and the proportion of nuclei p65, suggesting that JHQG treatment inhibited TLR4/MyD88/NF-κB pathway. CONCLUSION: JHQG reduced pulmonary inflammation and protected mice from LPS-induced ALI by promoting neutrophil apoptosis and inhibition of TLR4/MyD88/NF-κB pathway, suggesting that JHQG may be a promising drug for treatment of ALI.

HIV-1 Drug Resistance Profiles of Low-Level Viremia Patients and Factors Associated With the Treatment Effect of ART-Treated Patients: A Cross-Sectional Study in Jiangsu, China.

Objectives: Evaluating the drug resistance (DR) profiles of LLV patients and the influencing factors of treatment effects in Jiangsu Province. Method: The Pol gene (Reverse transcriptase and protease) was amplified and sequenced to identify the genotypes and DR profiles among LLV patients in 2021. Questionnaire survey was conducted among HIV/AIDS patients to investigate the potential influence factors of treatment effects. Results: 242 Pol genes were amplified from 345 specimens, and ten genotypes were detected. The DR rate was 40.5%, with 66, 86, and 14 being resistant to NRTIs, NNRTIs, and PIs, respectively. Patients treated with the 2NRTIs+PIs regimen were detected with more DR; and drug resistance was less detected in married or cohabiting patients than unmarried patients. Non-smokers were less likely to develop LLV at follow-up than smokers; patients with stage II clinical stage at diagnosis and using 2NRTIs+PIs regimen were also more likely to develop LLV at follow-up. Conclusion: Drug resistance profiles in LLV patients are severe and differ in treatment regimens and marital statuses. Meanwhile, smoking history, clinical stage, and treatment regimen may influence the therapeutic effect. It is necessary to include LLV people in the free drug resistance testing program.

Acceptability of HIV pre-exposure prophylaxis in China: A systematic review and meta-analysis.

Using pre-exposure prophylaxis (PrEP) for the prevention of HIV infection is a popular, contemporary research topic. We retrieved PubMed, Web of Science, MEDLINE, Wanfang, CNKI, and Sinomed databases, and related websites to identify studies both in Chinese and English, which reported the acceptability of PrEP in China. A total of 3203 citations were retrieved, of which 54 were included. We found that the acceptability varied widely across the country, ranging from 19.1% to 94.6%, and the pooled estimate was 66.8% (95% CI: 62.0%-71.3%). Higher acceptability of PrEP was associated with the following factors: individuals with a lower educational degree, higher income, non-discriminatory attitude towards AIDS or AIDS patients, self-perceived high HIV risk, feeling that PrEP should be promoted, familiarity with PrEP, history of HIV testing, history of anal intercourse, consistent condom use, higher number of sexual partners. The main reasons for not using PrEP are doubting its validity, fear of potential side effects and financial burden, and fear of stigma for using PrEP. This review found that the acceptance in China was moderate.

[Effect of moxibustion at "oppositely-located points" on neurogenic bladder after spinal cord injury and endoplasmic reticulum stress pathway in rats].

OBJECTIVE: To observe the effect of moxibustion at oppositely-located points "Mingmen" (GV 4) and "Shenque" (CV 8) on the motor function of the hind limbs and bladder function in rats with neurogenic bladder after suprasacral spinal cord injury (SCI), so as to explore the effect of this therapy on bladder tissue apoptosis mediated by endoplasmic reticulum stress pathway. METHODS: Twenty-eight female Wistar rats were randomly divided into a sham-operation group (8 rats) and a model establishment group (20 rats). Using the modified Allen's method, the spinal cord of T10 segment was injured to establish a neurogenic bladder model in the model establishment group. Sixteen rats were modeled successfully and then divided into a model group (8 rats) and a moxibustion group (8 rats). In the moxibustion group, 2 h after consciousness regaining from modeling anesthesia, moxibustion was exerted at "Shenque" (CV 8) and "Mingmen" (GV 4), 2 cones at each acupoint in one intervention. The intervention was administered once every two days and 5-time intervention was required totally. After intervention, Basso, Beattie and Bresnahan locomotor rating scale (BBB) score for the motor function of the hind limbs, and the urodynamics indexes (maximum bladder capacity, urine leakage pressure and bladder compliance) were compared among groups. HE staining method was adopted to observe the morphological changes of bladder tissue. With Western blot method and real-time PCR assay, the protein and mRNA expressions of the endoplasmic reticulum stress-related genes (glucose- regulated protein 78 [GRP78], activating transcription factor 4 [ATF4] and cysteinyl aspartate specific proteinase-12 [Caspase-12]) were determined. RESULTS: The transitional epithelial cells were arranged irregularly, the bladder wall was getting thinner, and the cellular vacuolar degeneration and neutrophil infiltration were found in the model group. Whereas, compared with the model group, in the moxibustion group, the arrangement of transitional epithelial cells was clear and continuous in layers, the cellular vacuolar degeneration was mild and the infiltration presented in a small amount of neutrophil granulocytes. Compared with the sham-operation group, in the model group, the BBB score was reduced (P<0.01), the maximum bladder capacity and bladder compliance were increased (P<0.01), and the protein expression levels of GRP78, ATF4 and Caspase-12, as well as mRNA expressions were all increased (P<0.01). In comparison with the model group, in the moxibustion group, BBB score was increased (P<0.01), the maximum bladder capacity and bladder compliance were decreased (P<0.01), and the protein and mRNA expression levels of GRP78, ATF4 and Caspase-12 were all decreased (P<0.01). CONCLUSION: Moxibustion at the "oppositely-located points" improves the urination function, alleviate urine retention in neurogenic bladder rats after spinal cord injury. The underlying mechanism may be related to the down-regulation of the expressions of GRP78, ATF4 and Caspase-12 in the endoplasmic reticulum stress pathway of the bladder tissues, and thus to alleviate the apoptosis of bladder tissue.

Short-term effects of lifestyle intervention in the reversion to normoglycemia in people with prediabetes.

AIMS: To evaluate the short-term effect of lifestyle intervention in people with prediabetes. METHODS: A stratified multistage sampling method was used in the recruitment of residents of the Jiangsu Province, China in 2017, who had no previous diagnosis of diabetes. Physical examination and laboratory tests were performed, and questionnaires were completed. Those with a prediabetes diagnosis at baseline were included in the cohort and participants were randomized to the intervention group or the control group. The intervention group received a lifestyle intervention strategy, which included exercise, diet and peer educations. The control group received general health education. Participants were followed up in 2018. RESULTS: A total of 2005 individuals were included in the analysis. At follow-up, there were 516 (36.7%) individuals in the intervention group and 207 (34.5%) individuals in the control group with normal blood glucose levels. The decline in waist circumference and fasting plasma glucose levels was significantly higher in the intervention group than in the control group. This was still observed after adjusting for variables (odds ratio 1.32, P = 0.02). Females or younger individuals who had lower body mass index and plasma glucose levels at baseline were more likely to reverse to normoglycemia at follow-up. CONCLUSIONS: Compared with a strategy of general health education, a lifestyle intervention strategy could reverse glucose levels to normoglycemia in individuals with prediabetes.

LncRNA LINC00461 Promotes Colorectal Cancer Progression via miRNA-323b-3p/NFIB Axis.

BACKGROUND: LncRNA LINC00461 has been reported to play crucial regulatory roles in a variety of biological processes, including cell migration, cell invasion and cancer progression. However, its biological role in colorectal cancer (CRC) is completely unknown. The aim of our study was to explore the function of LINC00461 on CRC cells and the underlying mechanism. MATERIALS AND METHODS: CRC tumor tissues and cell lines derived from hospital and corporation. The expression level of LINC00461 in CRC tissues and cell lines were analyzed by quantitative real-time PCR (qRT-PCR). The effect of LINC00461 on cell proliferation, colony formation, migration and invasion were detected by CCK-8 assay, colony formation and transwell assay, respectively. In addition, cell apoptosis was analyzed by flow cytometry, and the role of LINC00461 on tumor growth was investigated by tumor xenografts in nude mice. The targets of LINC00461 were predicted by starBase v3.0 and confirmed by a dual-luciferase reporter system. The expression level of transcription factors of nuclear factor I B (NFIB), p21 and CDK2 was determined by Western blot or qRT-PCR. The NFIB expression levels in CRC tissues and mice tumors were analyzed by immunofluorescence assay (IHC). RESULTS: We found that the expression of LINC00461 was significantly overexpressed in CRC tissues and different cell lines, and the high level of LINC00461 expression was associated with poor overall survival. Downregulation of LINC00461 expression significantly suppressed the proliferation, migration and invasion of CRC cells and promoted cell apoptosis. We also found that LINC00461 could directly interact with miR-323b-3p. In addition, LINC00461 significantly increased the expression NFIB and CDK2, but, p21 was inhibited. Finally, we found that the growth of tumors in nude mice was suppressed upon LINC00461 deletion. CONCLUSION: We demonstrated that LINC00461 may play an oncogenic role in CRC cells through NFIB signaling pathway by targeting miR-323b-3p. Our report showed that LINC00461 may be a prognostic biomarker and candidate therapeutic target for CRC.

Characteristics of the vaginal microbiome in cross-border female sex workers in China: a case-control study.

BACKGROUND: Female sex workers (FSWs) are key groups in the transmission of sexual transmitted infections (STI), and vaginal microbiome variations play an important role in transmission. We aimed to explore the characteristics of vaginal microbiome among FSWs. MATERIALS AND METHODS: A total of 24 cross-border FSWs were randomly selected from a cross-sectional survey for female sex workers in southwest China. Thirty-seven female non-sex workers (FNSWs) were randomly selected from the gynecology clinic and health examination center. Vaginal swabs were collected, bacterial DNA extracted and 16S rRNA genes were sequenced. Differences in the vaginal microbiome between both groups were compared using bioinformatics analysis. RESULTS: One DNA sample was excluded due to unqualified concentration, therefore 60 samples were sequenced. FSWs had significantly different vaginal microbiota ß diversity, but undifferentiated α diversity when compared with non-sex workers. The average relative abundance of Sneathia, Shigella, Neisseria, Chlamydia, Prevotella, Enterococcus and Ureaplasma among FSWs was higher than FNSWs, and relative abundance of Atopobium in FSWs was lower than FNSWs. The Lactobacillus genus was the major genus in both groups. At the species level, Lactobacllus crispatus, Lactobacllus gasseri and Lactobacllus jensenii, in female sex workers, were lower when compared to FNSWs. CONCLUSION: There were distinct differences in vaginal bacteria variety between FSWs and FNSWs. Some disease-related genus were also more abundant in FSWs. Based on these observations, further research is required to identify microbiome communities related to high STI risks and other diseases in these cohorts.