Objective: To evaluate the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) on the expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA) tissues. Methods: A total of 56 patients with CA were enrolled from Department of Dermatology, The First Affiliated Hospital of Zhengzhou University from October 2016 to September 2017, and skin lesions were obtained before and 1 week after the first ALA-PDT treatment. Immunohistochemical SP method was used to determine the expression of VEGF and PCNA in keratinocytes in the CA tissues. Chi-square test and rank sum test were carried out to analyze differences between pre- and post-treatment expression rate and intensity of VEGF and PCNA, and Spearman correlation analysis was conducted to analyze the correlations between the protein expression of VEGF and PCNA. Results: The expression rates of VEGF and PCNA in keratinocytes in the CA tissues were 71.43% (40/56) and 73.21% (41/56) respectively before ALA-PDT, and 44.64% (25/56) and 41.07% (23/46) respectively after ALA-PDT. There were significant differences between pre- and post-treatment expression rate and intensity of VEGF and PCNA (expression rate: χ² = 8.25, 11.81 respectively, both P < 0.05; expression intensity: H = 11.29, 12.22 respectively, both P < 0.05) . The expression of VEGF was positively correlated with the expression of PCNA in the CA tissues before and after the ALA-PDT treatment (r_s = 0.202, 0.273, respectively, both P < 0.05) . Conclusion The expression of VEGF and PCNA decreased in CA tissues after ALA-PDT treatment, which may be one of the mechanisms underlying the treatment of CA with ALA-PDT.

OBJECTIVETo observe the inhibitory effect of citrus extract nobiletin on K562 cells xenograft in nude mice and discuss its anticancer activity and mechanism.

METHODThe model of K562 cells xenograft was established in nude mice. Twenty-five nude mice were divided to five groups. After 24 hrs of inoculation with K562 tumor cells subcutaneously, 1% CMC-Na in the nude mice of model control group, nobiletin (12.5, 25, 50 mg x kg(-1)) in the nude mice of nobiletin groups and CTX (20 mg x kg(-1)) in positive control group were administered once every day. The nude mice were killed at 18th day-point of administration. The inhibitory rate of nobiletin on tumor was calculated according to the measured tumor weight. Immunohistochemistry assay was used to determine the effect of nobiletin on VEGF expression and MVD, and CAM assay was used to detect the effect of nobiletin on vessel regeneration.

RESULTNobiletin have notable inhibition on K562 cells xenograft in nude mice comparing with model control group (P < 0.01), the inhibitory rate of nobiletin groups were 36% -58%. The results of immunohistochemical technology showed that the expression of VEGF in nobiletin groups decreased significantly comparing with the model control group (P < 0.05, P < 0.01, P < 0.01). Nobiletin could remarkably decrease the angiogenesis within tumor tissues. The expression of CD34 in nobiletin low dose group and high dose group was lower than that in model control group (P < 0.05, P < 0.01). The result of CAM indicated that 4 microg and 2 microg nobiletin could inhibit the new blood vessels of CAM (P < 0.01).

CONCLUSIONNobiletin inhibited the tumor growth and angiogenesis by reducing the VEGF expression of K562 cells xenograft in nude mice.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Citrus ; chemistry ; Disease Models, Animal ; Flavones ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; K562 Cells ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms ; drug therapy ; genetics ; metabolism ; Transplantation, Heterologous ; Vascular Endothelial Growth Factors ; genetics ; metabolism

Objective To evaluate the safety and efficacy of haploidentical hematopoietic stem cell transplan-tation(haplo-HSCT)treatment in children with hematological diseases.Methods Fifty-nine cases of less than 14 years old children with hematonosis were analyzed retrospectively,who were enrolled in the Aerospace Central Hospital from July 2012 to June 2016.And the evaluation was carried out by analyzing the success rate of implantation,occu-rrence rate of graft versus host disease(GVHD),infection rate and transplant related mortality(TRM),cumulative re-currence rate,overall survival rate(OS)and disease-free survival rate(DFS).Results In total of 59 cases,the 59 engraftments were successfully transplanted,the median time of leukocyte engraftment was 18(8-23)days,the median time of platelet engraftment was 21(11-68)days,the bone marrow was assessed 28 days after transplanta-tion,which showed that 59 patients achieved complete remission(CR)and DNA test confirmed complete donor chime-rism.With a median of follow-up time of 19(5-56)months,the cumulative recurrence rates ofⅠ,Ⅱgrade andⅢ,Ⅳ grade acute GVHD were(38.3 ± 6.3)%(23 cases)and(16.7 ± 4.8)%(10 cases),respectively,the chronic GVHD cumulative recurrence rate was(65.6 ± 7.5)%(30 cases),the cytomegalovirus(CMV)viremia cumulative recurrence rate was(45.1 ± 6.5)%(27 cases),the Epstein-Barr virus(EBV)viremia cumulative recurrence rate was(10.0 ± 3.9)%(6 cases),the viral cystitis cumulative recurrence rate was(20.0 ± 5.5)%(12 cases),the transplant related mortality was(12.8 ± 6.0)%,the 2-year cumulative recurrence rate of CR group was(8.0 ± 5.4)%,and that of non-remission(NR)group was(64.1 ± 11.9)%.The 2-year OS of CR group was(78.9 ± 7.5)%,the 2-year OS of NR group was(32.5 ± 12.9)%,the 2-year DFS of CR group was(79.5 ± 9.8)%,the 2 years DFS of NR group was(27.4 ± 7.9)%.Conclusions Haplo-HSCT is safe and effective in treating children with hematonosis,and haplo-HSCT has high survival rate and low recurrent,especially when transplantation is per-formed in the remission stage.But the prognosis of haplo-HSCT is poor in the refractory and relapsed patients,and to explore the preventing recurrence measures are very urgent.

Objective To analyze the clinical features and imaging findings of lesser trochanter osteoidosteoma,and to discuss the causes of its misdiagnosis as chronic osteomyelitis.Methods The clinical features,X-ray,CT and MRI findings of 6 cases with pathologically confirmed osteoidosteoma in the lesser trochanter were reviewed retrospectively.Symptoms included knee pain (1 case),thigh pain (4 cases)and hip joint pain(1 case);claudication(2 cases),and night pain(1 case).Five patients had right-side,and 1 patient had left-side involvement.All the 6 cases were misdiagnosed as chronic osteomyelitis before operation.Results Four patients had CT scan,which showed intra-cortical niduses at the lesser trochanters with peri-focal sclerosis,joint capsule swelling and joint effusion. Five patients had MRI exams,MR images showed bone marrow edema,synovial thickening,joint capsule swelling and joint effusion in all the 5 cases,but only 2 showed niduses.Six patients had X-ray imaging exams,X-ray images showed bone sclerosis without radiolucent nidus.Conclusion Osteoidosteoma in the lesser trochanter may display atypical clinical features that might be difficult to be differentiated from chronic osteomyelitis without sufficient examination.CT is best in showing niduses,except some niduses with atypical shape,superficial location and high-density calcification.MRI-T2WI is sensitive in showing the inflammation and bone marrow edema with high signal intensity,which may affect nidus displaying.X-ray images can only display bone sclerosis without niduses.

OBJECTIVE: To explore whether MRI fusion technology (combined T2-weighted imaging [T2WI] and fat-suppressed T2WI [T2WI-(FS)]) improves signal differences between anal fistulas and surrounding structures. MATERIALS AND METHODS: A total of 32 patients with confirmed diagnoses of anal fistula were retrospectively studied. All available T2WI and T2WI-(FS) images for each patient were used to generate fusion image (T2WI-(Fusion)) based on the addition of gray values obtained from each pixel via an MR post-processing work station. The discriminability of fistula, perianal sphincter, and perianal fat in T2WI, T2WI-(FS), and T2WI-(Fusion) images was quantified with Fisher's scoring algorithm. For subjective visual image assessment by researchers, five-point scale scores were determined using a modified double-stimulus continuous quality-scale test to evaluate T2WI-(FS), T2WI, enhanced axial three-dimensional-volumetric interpolated breath-hold examination (3D-VIBE), and T2WI-(Fusion) sequence images. The differences were subsequently compared. RESULTS: Mean Fisher scores for fistulas vs. sphincters obtained from T2WI-(Fusion) (F(Fusion-fistula) = 6.56) were significantly higher than those from T2WI (F(T2WI-fistula) = 3.35) (p = 0.001). Mean Fisher scores for sphincters vs. fat from T2WI-(Fusion) (F(Fusion-sphincter) = 10.84) were significantly higher than those from T2WI-(FS) (FS(FS-sphincter) = 2.57) (p = 0.001). In human assessment, T2WI-(Fusion) showed the same fistula discriminability as T2WI-(FS), and better sphincter discriminability than T2WI. Overall, T2WI-(Fusion) showed better discriminability than T2WI, T2WI-(FS), and enhanced 3D-VIBE images. CONCLUSION: T2WI and T2WI-(FS) fusion technology improves signal differences between anal fistulas and surrounding structures, and may facilitate better evaluation of anal fistulas and sphincters.
Anal Canal ; Diagnosis ; Fistula ; Humans ; Magnetic Resonance Imaging ; Rectal Fistula ; Retrospective Studies

Objective:To analyze the genetic characteristics of the complete genome of a strain of human respiratory syncytial virus (HRSV) in Qinghai province in 2024.Methods:A total of 300 samples were collected during 2024 influenza surveillance in Qinghai province sentinel hospitals from patients with fever accompanied by severe respiratory infection symptoms. We used real-time fluorescent quantitative reverse transcription polymerase chain reaction RT-PCR) method to screen out HRSV subtype B (HRSVB) positive specimens, whole genome sequencing was performed on positivespecimens meeting the requirements for the sequencing. After downloading the global representative HRSVB genotypes at GenBank database, sequence alignment was performed, related evolutionary tree was built and the calculation and analyses of genetic distance were done, analyses of HRSVB sequencing of sequence homology of nucleotides, amino acids and amino acid mutation were performed.Results:The first strain in Qinghai, China/qinghai/2024-03 had a complete sequence of 15 140 bp nucleotides, with HRSV′s all structural characteristics, and subtype HRSVA prototype strain Long strains of nucleotide the lowest homology was 80.0%, and subtype HRSVB prototype strain nucleotide homology was above 94.7%. The result indicated that the first strain in Qinghai belonged to HRSVB subtype. Genetic evolution shows China/qinghai/2024-03 and USA/WA-S23450/2021 (OR326803.1) and Germany/2021 (OR795235.1) all belong to a branch, they have the closest relationship. Phylogenetic analysis of G gene showed that the strain belonged to BA9 genotype of HRSVB subtype, and the hypervariable regions of the genome were SH and G genes.Conclusions:In this study, the complete genome sequence of HRSV China/qinghai/2024-03 was obtained for the first time, and the basic molecular structural characteristics were elucidated, which filled the gaps in the gene and amino acid data of HRSV in our province, and also provided a basis for HRSV epidemiology.

BACKGROUND:Cannabidiol is effective in ameliorating the body's inflammatory response,but no clear mechanistic studies have been conducted to ameliorate skeletal muscle inflammation induced by exhaustive exercise. OBJECTIVE:To explore the mechanism by which cannabidiol improves skeletal muscle inflammation during exhaustive exercise by using transcriptome sequencing technology. METHODS:Thirty-six Sprague-Dawley rats were randomly divided into six groups:blank control group,exercise coconut oil group,exercise control group,50 mg/kg cannabidiol group,60 mg/kg cannabidiol group,and 70 mg/kg cannabidiol group,with six rats in each group.Except for rats in the blank control group,rats in each group were subjected to swimming exercise for 9 days to produce the exhaustive exercise model.At the end of each swimming exercise,rats in the cannabidiol groups were given 2 mL of fat-soluble cannabidiol at different concentrations(50,60,and 70 mg/kg)by gavage;rats in the exercise coconut oil group were given the same volume of coconut oil by gavage until the end of the exercise on the 9th day;and rats in the blank control group and the exercise control group were not given any special treatment.The levels of inflammatory factors and differentially expressed genes in the skeletal muscle of rats in each group were determined using ELISA and transcriptome sequencing techniques.Differentially expressed genes obtained were subjected to KEGG analysis,and the accuracy of the sequencing data was verified by fluorescence quantitative PCR. RESULTS AND CONCLUSION:The results of ELISA showed that the contents of interleukin-6(P<0.05),tumor necrosis factor-α(P<0.01),interleukin-10 and other inflammatory factors in the exercise group increased significantly compared with the blank control group and the coconut oil group.After cannabidiol intervention,the mass concentrations of interleukin-6 and tumor necrosis factor-α showed a sequential decrease with increasing cannabidiol concentration.By comparing GO and KEGG databases,the functional properties of differentially expressed genes were analyzed,and the results showed that the differentially expressed genes were mainly involved in the tumor necrosis factor signaling pathway and the Toll-like receptor signaling pathway.RT-qPCR results showed that the trends of five randomly selected differentially expressed genes were in agreement with the transcriptome sequencing results.To conclude,cannabidiol can improve skeletal muscle inflammation caused by exhaustive exercise.

Glycosphingolipids (GSLs) are widely distributed in the phospholipid bilayer of various cell membranes, which play an important role in maintaining cell membrane stability, and regulate various cellular processes including adhesion, proliferation, apoptosis and recognition, as well as participate in various cellular activities. In addition, GSLs are not only involved in the process of apoptosis, but also regulate multiple signals in tumorigenesis and tumor development. The tumor-associated GSLs are expected to be used as diagnostic markers and immunotherapeutic targets for malignant tumors. These findings have important implications for the study of apoptosis and provide the new direction of tumor therapy. This review summarized the latest research progress of GSLs-mediated apoptosis and its effect on the genesis, development and metastasis of tumor cells. Moreover, we discussed the metabolic pathway of GSLs-mediated apoptosis and its application in tumor therapy, as well as the development prospect of targeted therapy strategies based on GSLs.
Humans ; Glycosphingolipids/metabolism* ; Apoptosis ; Cell Membrane ; Neoplasms/metabolism*

Objective:The two detection method for the detection of hepatitis A virus (HAV) in strawberries were optimized and compared to select the best detection method for the detection of hepatitis A virus in strawberries.Methods:Different concentrations of HAV were inoculated on the surface of known negative frozen strawberry specimens, the concentration of beef extract powder in alkaline elution-PEG concentration method was optimized, the optimal nucleic acid extraction kit was selected, the optimal lysis buffer volume in direct lysis method was optimized, and the recovered viral load was detected by Real-time fluorescence quantitative RT-PCR. SPSS26.0 was used to statistically analyze the data, and the optimized two method were used for the detection of actual specimens.Results:The concentration of beef extract powder by the optimized alkaline elution-PEG concentration method was selected at 3%, and the viral nucleic acid extraction kit was selected as kit B. Six ml of lysis buffer was selected for the optimized direct lysis method. The recovery rates of HAV virus by alkaline elution-PEG concentration and direct lysis were compared with the HAV addition levels, and the HAV virus recovery rates of the two method were 21.50±1.06% and 5.82±0.01%, respectively, and the result showed that the differences were statistically significant. A total of 60 strawberry specimens from four regions were tested at the same time, and the result were all negative.Conclusions:The optimized alkaline elution-PEG concentration method has higher sensitivity and is more suitable for the detection of HAV in strawberry specimens.

Objective:To establish a digital droplet RT-PCR(dRT-PCR) method for Hepatitis A virus (HAV), and compare it with Real time RT-PCR(RT-qPCR) method, and select the best method for detecting hepatitis A virus in strawberry.Methods:Extract HAV vaccine RNA, optimize the reaction conditions of dRT-PCR and evaluate its specificity; Alkaline elution -PEG concentration method was used to extract nucleic acid from strawberry samples. At the same time, dRT-PCR and RT-qPCR method were used to detect the sensitivity and inhibition rate of HAV vaccine RNA in pure water and strawberry matrix, and the recovery rate of HAV in artificially contaminated strawberry was compared, which was applied to the detection of commercially available samples.Results:The optimal annealing temperature for dRT-PCR reaction was 60 ℃, and the optimal concentrations of primers and probes were 0.4 μmol/L、0.4 μmol/L and 0.2 μmol/L, with good specificity. There is no significant difference in sensitivity between the two method in detecting HAV vaccine RNA in pure water and strawberry matrix. The inhibition rate of dRT-PCR is low. The recovery rates of dRT-PCR and RT-qRCR in the detection of strawberry samples contaminated with HAV at higher concentrations were 12.90±0.006% and 30.12±0.02%, respectively. The recovery rates of lower concentrations of HAV contaminated strawberry samples were 18.27±0.07% and 10.85±0.03%, respectively, and the difference was statistically significant ( P<0.05). When strawberry samples on the market were tested, the result of both method were negative. Conclusions:The sensitivity of dRT-PCR method established in this study is not significantly different from that of RT-qPCR in detecting HAV RNA in different substrates, but dRT-PCR has good tolerance to PCR reaction inhibitors and high recovery rate when detecting low concentration HAV. Both detection method can be used for quantitative detection of hepatitis A virus in strawberry, and can be selected according to the actual situation.