Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

Objective:To discuss the effect of downregulating microRNA-208a(miR-208a)on the resistance of the colorectal cancer cells to 5-fluorouracil(5-FU),and to clarify its related molecular mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-208a and secreted frizzled-related protein 1(SFRP1)mRNA in the 5-FU-resistant colorectal cancer cell line HT-29/5-FU and its parent HT-29 cells.The HT-29/5-FU cells were transfected with miR-208a inhibitor plasmid and its negative control plasmid(inhibitor-NC),and SFRP1 small interfering RNA(si-SFRP1)and its negative control plasmid(si-NC),either separately or in combination,followed by treatment with 5-FU.The cells were divided into inhibitor-NC group,miR-208a inhibitor group,miR-208a inhibitor+si-NC group,and miR-208a inhibitor+si-SFRP1 group.MTT assay was used to detect the proliferation activities of the cells and the resistance indexes were calculated;Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptotic rates of the cells after treated with different concentrations of 5-FU;Western blotting method was used to detect the expression levels of SFRP1,β-catenin,P-glycoprotein(P-gp),and ATP-binding cassette subfamily B member 1(ABCB1)proteins in the cells in various groups;dual-luciferase reporter gene assay was used to validate the targeting relationship between miR-208a and SFRP1.Results:Compared with HT-29 cells,the expression level of miR-208a in the HT-29/5-FU cells was increased(P＜0.05),and the expression level of SFRP1 mRNA was decreased(P＜0.05).Compared with inhibitor-NC group,the proliferation activity of the cells in miR-208a inhibitor group was decreased(P＜0.05),the resistance index was decreased,the apoptotic rate was increased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were decreased(P＜0.05).The dual-luciferase reporter gene assay results showed that SFRP1 was a target gene of miR-208a and miR-208a could negatively regulate the expression of SFRP1.Compared with miR-208a inhibitor+si-NC group,the proliferation activity of the cells in miR-208a inhibitor+si-SFRP1 group was increased(P＜0.05),the resistance index was increased,the apoptotic rate was decreased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were increased(P＜0.05).Conclusion:Downregulation of miR-208a can improve the resistance of the HT-29/5-FU cells to 5-FU by targeting and upregulating the SFRP1 expression,thereby inhibiting the transmission of the Wnt signaling pathway.

Colorectal cancer (CRC) is a type of malignant tumor that seriously threatens human health and life, and its treatment has always been a difficulty and hotspot in research. Herein, this study for the first time reports that antipsychotic aripiprazole (Ari) against the proliferation of CRC cells both in vitro and in vivo, but with less damage in normal colon cells. Mechanistically, the results showed that 5-hydroxytryptamine 2B receptor (HTR2B) and its coupling protein G protein subunit alpha q (Gαq) were highly distributed in CRC cells. Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling. Blockade of HTR2B signaling suppressed the growth of CRC cells, but HTR2B was not found to have independent anticancer activity. Interestingly, the binding of Gαq to HTR2B was decreased after Ari treatment. Knockdown of Gαq not only restricted CRC cell growth, but also directly affected the anti-CRC efficacy of Ari. Moreover, an interaction between Ari and Gαq was found in that the mutation at amino acid 190 of Gαq reduced the efficacy of Ari. Thus, these results confirm that Gαq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation. Collectively, this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.

OBJECT IVE To study the effects and potential mechanism of saltwater stir-baked Eucommia ulmoides on kidney-yang deficiency in rats. METHODS Ninety SD rats were randomly divided into normal control group （15 rats） and modeling group （75 rats）. Modeling group was given adenine intragastrically to establish kidney-yang deficiency model. After modeling，modeling group were randomly divided into model group ，positive control group （Guifu dihuang tablet 2.5 g/kg）， low-dose，medium-dose and high-dose groups of saltwater stir-baked E. ulmoides （1.5，3，6 g/kg），with 15 rats in each group. Administration groups were given relevant medicine intragastrically ，normal control group and model group were given normal saline intragastrically ，once a day ，for consecutive 8 weeks. The body weight of rats was measured before modeling and after medication. The score of kidney-yang deficiency syndrome was performed after medication. The levels of blood urea nitrogen （BUN），serum creatinine （SCR），testosterone（T）and cortisol （COR）in serum and superoxide dismutase （SOD）activity， malondialdehyde（MDA）level in renal tissue were detected. The pathological changes of renal tissue were observed after HE staining. Relative expressions of hypoxia-inducible factor- 1 α（HIF-1 α）and signal transducer and activator of transcription 5 （STAT5） mRNA in renal tissue were detected by RT-PCR. The relative expressions of HIF- 1α，STAT5 and phosphorylated STAT 5 （p-STAT5）protein and the ratio of gray values of p-STAT 5 and STAT 5（p-STAT5/STAT5）in renal tissue were detected by Western blot. RESULTS Before modeling ，there was no statistical significance in body weight of rats in each group （P＞0.05）. After medication， compared with model group ， pathological changes of renal tissue were all recovered in saltwater stir-baked E. ulmoides groups and positive control group ， while body weight ，the level of T in serum and SOD activity qq.com in renal tissue were all increase d significantly （P＜0.05）. The scores of kidney-yang deficiency syndrome ，levels of BUN ， SCR and COR in serum ，MDA level in renal tissue ，the relative expressions of HIF- 1α and STAT5 mRNA，the relative expressions of HIF- 1α，STAT5 and p-STAT 5 protein as well as p-STAT5/STAT5 were all significantly decre ased （P＜0.05）. The above effects of saltwater stir-baked E. ulmoides were in dose-dependent manner （P＜0.05）. CONCLUSIONS Saltwater stir-baked E. ulmoides can significantly relieve renal tissue damage in rats with kidney-yang deficiency ，decrease the levels of BUN ，SCR and COR in serum ，increase the level of T in serum ，the mechanism of which may be associated with anti-oxidative stress and inhibition of the expression of HIF- 1α and STAT5 protein.

It has been demonstrated that APPL1 (adaptor protein, phosphotyrosine interacting with PH domain and leucine zipper 1) is involved in the regulation of several growth-related signaling pathways and thus closely associated with the development and progression of some cancers. Diallyl trisulfide (DAT), a garlic-derived bioactive compound, exerts selective cytotoxicity to various human cancer cells through interfering with pro-survival signaling pathways. However, whether and how DAT affects survival of human hepatocellular carcinoma (HCC) cells remain unclear. Herein, we tested the hypothesis of the involvement of APPL1 in DAT-induced cytotoxicity in HCC HepG2 cells. We found that Lys 63 (K63)-linked polyubiquitination of APPL1 was significantly decreased whereas phosphorylation of APPL1 at serine residues remained unchanged in DAT-treated HepG2 cells. Compared with wild-type APPL1, overexpression of APPL1 K63R mutant dramatically increased cell apoptosis and mitigated cell survival, along with a reduction of phosphorylation of STAT3, Akt, and Erk1/2. In addition, DAT administration markedly reduced protein levels of intracellular TNF receptor-associated factor 6 (TRAF6). Genetic inhibition of TRAF6 decreased K63-linked polyubiquitination of APPL1. Moreover, the cytotoxicity impacts of DAT on HepG2 cells were greatly attenuated by overexpression of wild-type APPL1. Taken together, these results suggest that APPL1 polyubiquitination probably mediates the inhibitory effects of DAT on survival of HepG2 cells by modulating STAT3, Akt, and Erk1/2 pathways.

Objective:To evaluate the impact of sarcopenia on survival and treatment-related toxicity in postoperative recurrent esophageal squamous cell carcinoma (ESCC) patients treated with chemoradiotherapy.Methods:Clinical data of 147 patients with postoperative locoregional recurrent ESCC receiving chemoradiotherapy in Huai'an First People's Hospital from 2016 to 2017 were retrospectively analyzed. Pectoralis muscle area (PMA) was determined using routine pre-radiotherapy CT simulation scan above the aortic arch level. Sarcopenia was defined as a cut-off value of pectoralis muscle index (PMI) (PMA/height 2) <11.55 cm 2/m 2 for males and <8.69 cm 2/m 2 for females. The incidence of toxicity, 1- and 3-year overall survival (OS) rates were statistically compared between patients with and without sarcopenia. Results:Sarcopenia was detected in 49 of 147 (33.3%) patients. The incidence of grade 3-4 toxicities in sarcopenic patients was significantly higher compared to that in their counterparts without sarcopenia (40.8% vs. 18.4%, P=0.005). In addition, patients with sarcopenia had significantly worse 1-year (61.2% vs. 82.7%) and 3-year OS rates (10.2% vs. 28.6%) than those without sarcopenia (both , P<0.001). Multivariate analysis showed that sarcopenia was an independent prognostic factor for poor OS ( P<0.001). Conclusion:PMI based on CT simulation scan has prognostic value in postoperative locoregional recurrent ESCC patients treated with chemoradiotherapy, which probably serves as a novel diagnostic tool for sarcopenia.

Objective:To investigate the clinical and imaging characteristics of liver density changes in patients with initial-treated drug-sensitive secondary tuberculosis during standardized treatment and after withdrawal when cured.Methods:A retrospective analysis was conducted of 34 patients with initial-treated drug sensitive pulmonary tuberculosis in Beijing Chest Hospital of Capital Medical University and the First Affiliated Hospital of Xinxiang Medical University from January 2014 to April 2019. The chest computed tomography (CT) examination and sputum culture were performed before treatment. The patients received the standardized treatment and they were divided into three groups according to the course of treatment (three, nine and 12 months). Liver density and liver function were followed up during treatment (three, six, nine and 12 months) and after drug withdrawal (3, 6 and 12 months). The measurement data were analyzed by t-test. Results:The average liver density of these three groups gradually decreased during the treatment period, and gradually increased after drug withdrawal. There were five and nine cases of fatty liver occurred at three and six months of treatment in the six-month treatment group, respectively; and six, two and zero cases of fatty liver occurred at three, six, and 12 months after drug withdrawal, respectively. There were four, eight and 11 cases of fatty liver occurred at three, six, and nine months of treatment in the nine-month treatment group, respectively; and seven, two and zero cases occurred at three, six, and 12 months after drug withdrawal, respectively.There were five, 10, 14 and 14 cases of fatty liver occurred at three, six, nine and 12 months of treatment in the 12-month treatment group, respectively; and 12, 10 and five cases occurred at three, six, and 12 months after drug withdrawal, respectively. During the course of treatment, the density of livers of some cases decreased unevenly, and the density of right lobe of the liver was lower than the left lobe. The density of left lobe of the liver was (49.8±4.0) HU, (45.0±3.9) HU, (37.0±9.9) HU, (45.3±8.1) HU, (48.4±6.6) HU at the treatment of six, nine and 12 months and drug withdrawal of three and six months, and the density of right lobe of the liver was (44.0±6.1) HU, (37.2±7.7) HU, (25.5±15.8) HU, (38.5±11.7) HU, (43.8±9.9) HU, the differences were statistically significant ( t=4.611, 4.512, 2.307, 2.803 and 2.291, respectively, all P<0.05), while those were not statistically significant among three months of treatment and 12 months after drug withdrawal ( t=1.573 and 1.199, respectively, both P>0.05). There were two cases showed alanine aminotransferase (ALT) and aspartate amiotransferase (AST) accompanied elevated (ALT>2×upper limits of normal (ULN), AST<2×ULN) at three and six months of treatment, with no abnormalities detected of alkaline phosphatase (ALP) and total bilirubin (TBil). Conclusions:The liver density gradually decreases and uneven fatty liver could appear during anti-tuberculosis treatment, but it gradually returns to normal or relieves after drug withdrawal. The degree of fatty liver is not synchronized with the changes of liver function indexes (ALT, AST, ALP and TBil), which belongs to chronic reversible injury.

Mycophenolic acid (MPA, ) and its derivatives are first-line immunosuppressants used in organ transplantation and for treating autoimmune diseases. Despite chemical synthetic achievements, the biosynthetic formation of a seven-carbon carboxylic acid pharmacophore side chain of , especially the processes involving the cleavage of the prenyl side chain between DHMP () and DMMPA (), remains unknown. In this work, we identified a membrane-bound prenyltransferase, PgMpaA, that transfers FPP to to yield FDHMP (). Compound undergoes the first cleavage step a new globin-like enzyme PgMpaB to form a cryptic intermediate . Heterologous expression of genes in demonstrates that the second cleavage step (from to ) of is a cluster-independent process . Our results, especially the discovery of the broad tolerance of substrates recognized by PgMpaB, set up a strategy for the formation of "pseudo-isopentenyl" natural products using fungal globin-like enzymes.

Fungal polyketides display complex structures and variously biological activities. Their biosynthetic pathways generally contain novel enzyme-catalyzed reactions. This review provides a summary of recent research advances in molecular mechanism of the biosynthesis of fungal polyketides including highly-reducing polyketide synthases (HR-PKSs), non-reducing polyketide synthases (NR-PKSs), as well as polyketide-nonribosomal peptide synthase (PKS-NRPSs) and reducingnon- reducing polyketide synthase (HR-NR PKSs) hybrids. The elucidation of biosynthetic mechanism of many fungal polyketides provides guidance on the discovery of new biosynthetic gene cluster of fungal polyketide natural products and compounds with novel structures as well as their analogue.

Objective To investigate the correlation between tumor perfusion parameters and tumor volume and Child-Pugh classification in CT of hepatocellular carcinoma (HCC).Methods Fifty-six patients with hepatocellular carcinoma were selected to perform CT perfusion imaging.The parameters of the total tumor perfusion such as hepatic artery perfusion (HAP),portal vein perfusion (PVP)and hepatic perfusion index (HAPI)were calculated according to the degree of hepatic encephalopathy,albumin,bilirubin, clotting time,ascites for liver Child-Pugh classification.The relationship between the tumor or peritumoral perfusion parameters with tumor volume and Child-Pugh classification were analyzed.Results (1)There was no correlation between tumor or peritumoral perfusion with the tumor volume.(2)The difference of HAP,PVP and HAPI between the different Child-Pugh classification groups was statistically significant (P<0.000 1).(3)With the reduction of Child-Pugh classification,the tumor body HAP and HAPI values gradually decreased, while the PVP value increased gradually.Conclusion There is no correlation between the tumor volume of hepatocellular carcinoma with total tumor perfusion parameters.The differences in perfusion measurements between different Child-Pugh classification can intuitively and quantitatively reflect the reserve function of the liver.