Objective To investigate the protective effect of herba artemisiae scopariae extract on multidrug resistance protein 3(MDR3)gene mutation-induced neonatal parenteral nutrition-associated cholestasis(PNAC)and its possible mechanism.Methods ①Human primary hepatocytes were treated with cell culture in vitro,CRISPR/Cas9 lentivirus infection and MDR3 mutant gene lead-in.The levels of hepatic and biliary biochemical indexes[alanine transaminase(ALT),aspartate transaminase(AST),total bilirubin(TBil),direct bilirubin(DBil),indirect bilirubin(IBil),total bile acid(TBA)]in the supernatant of hepatocytes before and after 16,32,48 hours were compared to determine the time required for fatty acid induction of PNAC hepatocyte model with MDR3 gene mutation.② Human primary hepatocytes were divided into blank control group,MDR3 gene wild type group,MDR3 gene mutation group,and herba artemisiae scopariae extract intervention group according to random number table method.The blank control group was treated with culture medium only,the MDR3 gene wild type group was infected with lentivirus and mixed with wild type MDR3 gene and culture medium,the MDR3 gene mutation group was infected with lentivirus and cultured in culture medium with the mutant genes lead-in of LV-MDR3KI(c.485T>A,c.2793insA,c.1031G>A,c.3347G>A)mutation,while the MDR3 mutant gene was lead-in by lentivirus infection and cultured in culture medium,and then pretreated with 100 g/L herba artemisiae scopariae extract in the herba artemisiae scopariae extract intervention group,then the four groups of hepatocytes were induced with 1%fat emulsion,and the treatment time was the time needed to construct the PNAC hepatocytes model with MDR3 gene mutation.The levels of ALT,AST,TBil,DBil,IBil and TBA in the supernatant of hepatocytes were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression abundance of adenosine triphosphate binding cassette proteins(ABCB4,ABCB11,ABCC2,ABCC3,ABCC4)encoding MDR3,bile salt export pump(BSEP),multidrug resistance associated protein(MRP)2-4,and tumor necrosis factor-α(TNF-α)genes were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared to the blank control group and MDR3 gene wild type group,there was no significant difference in the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 gene mutant group before and 16 hours after induction with 1%fat emulsion,however after treated with 1%fat emulsion for 32 hours and 48 hours,the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 mutant hepatocytes were significantly increased(P<0.05),consequently the time required for fatty acid induction of PNAC hepatocyte model was 32 hours.At 32 hours after treatment with fat emulsion,the levels of ALT,AST,TBil,DBil,TBA in the supernatant of hepatocytes in the herba artemisiae scopariae extract intervention group were significantly decreased[ALT(ng/L):148.3±2.3 vs.164.9±7.0,AST(ng/L):2767.4±78.8 vs.3239.4±107.1,TBil(μmol/L):7.6±0.2 vs.13.6±0.3,DBil(μmol/L):1.8±0.1 vs.5.7±0.2,TBA(μmol/L):3.4±0.2 vs.6.7±0.1,all P<0.05].The ABCB4,ABCC2,ABCC3,ABCC4 mRNA expression of MDR3,MRP2,MRP3,MRP4 in the blank control group,MDR3 wild type group,MDR3 gene mutation group and the herba artemisiae scopariae extract intervention group had no significant difference.The expression of TNF gene mRNA was highly expressed in MDR3 gene mutation group(2-??Ct:1.258±0.200 vs.1.001±0.052),and was low expressed in the herba artemisiae scopariae extract intervention group(2-??Ct:0.387±0.247 vs.1.258±0.200),and there was a significant difference between the two groups(both P<0.05).Compared to the MDR3 gene mutation group,the ABCB11 gene encoding BSEP mRNA expression in the herba artemisiae scopariae extract intervention group was significantly increased(2-??Ct:2.955±0.479 vs.1.333±0.529,P<0.05).Conclusion The herba artemisiae scopariae extract has a protective effect on PNAC induced by MDR3 gene mutation,which may be related to antagonizing inflammatory reaction,decreasing the expression of TNF mRNA and improving the expression of ABCB11 gene encoding BSEP.

Objective:To analyze the clinical characteristics of monkeypox patients.Methods:The clinical data and laboratory findings of 4 patients with monkeypox patients diagnosed at Yiwu Central Hospital in July 2023 were analyzed. Herpes fluid and skin tissue samples were collected, the viruses were isolation and cultured in African green monkey kidney cells (Vero) and identified with whole gene sequencing.Results:All four patients were male, aged 24-35 years. All patients had male-to-male behavior within 21 days before onset of the disease. Among them, one patient has AIDS and one patient has syphilis. Four patients presented with perineal skin lesions with itching, and 3 patients were found to have enlarged lymph nodes upon admission. Laboratory testing: lymphocyte abnormality (4.57×10 9/L) in 1 case; increased procalcitonin (0.25 ng/mL) in 1 case; elevated IL-10 levels ( 7.11 ng/L and 9.42 ng/L) in 2 cases; increased IL-6 (66 ng/L) and IL-4 (3.24 ng/L) in 1 case, respectively. One case had abnormal myocardial zymogram with a elevated lactate dehydrogenase level of 313 U/L. The monkeypox virus was isolated from lesion tissue and herpes fluid, and the whole gene sequencing identified it as the B. 1.3 subtype of the IIb evolutionary branch, exhibiting typical pathological effects on Vero cells. Conclusion:The clinical manifestations of the 4 monkeypox patients confirmed in Zhejiang province are mild, patients had a definitive history of male-to-male sexual behavior and the virus strains belong to the B. 1.3 lineage of the IIb evolutionary branch.

COVID-19/virology* ; China/epidemiology* ; Disease Outbreaks ; Genetic Linkage ; Humans ; Phylogeny ; Polymorphism, Single Nucleotide ; SARS-CoV-2/isolation & purification*

Objective:To analyze Helicobacter pylori ( Hp) resistance in Xiaoshan area from 2013 to 2020 and to provide reference for the selection of clinical antibiotics for the eradication of Hp in this area. Methods:Non-repetitive Hp strains isolated from patients treated in the First People′s Hospital of Xiaoshan District, Hangzhou from January 2013 to December 2020 and the drug susceptibility test results of these strains were collected. The patients were grouped by every 10 years of age to analyze the drug resistance of Hp in different groups and its changing trends. Results:A total of 54 912 Hp strains were isolated and cultured. The resistance rates of Hp isolates to amoxicillin, metronidazole, clarithromycin and levofloxacin were 0.42%, 92.94%, 22.41% and 29.83%, respectively. The drug resistance rates to clarithromycin and levofloxacin were increasing year by year. The resistance rates to levofloxacin, clarithromycin and metronidazole increased with age. The resistance rates to clarithromycin, levofloxacin and metronidazole in women were higher than those in men, especially in the 21-60 age group. The resistance rate to clarithromycin was significantly higher in people under 20 years old. In 2017 to 2019, the resistance rate to amoxicillin was higher in people under 20 years old than in other age groups. In 2020, the resistance rates to levofloxacin and clarithromycin in people over 70 years old both exceed 57% and increased rapidly. Conclusions:The resistance rates of Hp to levofloxacin, clarithromycin and metronidazole in Xiaoshan area were all at a high level. It was recommended to use individualized treatments for the eradication of Hp. In addition, attention should be paid to the young and older people.

Objective:To explore the short-term effectiveness of arthroscopic combined with dual-plane high tibial osteotomy in the treatment of anterior cruciate ligament injury combined with varus deformity of knee joint.Methods:A retrospective study was performed on 17 patients with anterior cruciate ligament injury combined with varus deformity of knee joint who underwent arthroscopic combined with dual-plane high tibial osteotomy at Department of Bone and Joint, the Affiliated Hospital of Southwest Medical University from January 2017 to June 2018.There were 11 males (11 knees) and 6 females (6 knees), aged 41.3 years (range: 32 to 49 years) .During the surgery, the weight bearing line of lower extremity was set to 62.5% position of the tibial plateau on coronal plane. The tibial slope was adjusted to the normal range on sagittal plane, and anterior cruciate ligament was reconstructed to improve the stability of knee joint.At final follow up, full length weight bearing X ray was used to evaluate the position of weight bearing line, femoral tibial angle and tibial slope pre- and post-operatively.The Lysholm scores, Hospital for Special Surgery score, Tegner knee activity scores and International Knee Documentation Committee (IKDC) scores were used to estimate knee joint function, while the Lachman test, KT-1000 side-to-side difference and pivot-shift test were used to estimate the knee joint stability.Results:The patients were followed up for 1.8 years(range:1.2 to 2.5 years). No complication such as infection, deep vein thrombosis, graft failure, nonunion or delayed union was observed.The weight bearing line was corrected from (28.48±2.24)% preoperatively to (57.43±1.02)% postoperatively ( t=46.80, P=0.00) .The femoral tibial angle was improved from (172.31±3.37) ° preoperatively to (178.91±1.34) ° postoperatively( t=10.46, P=0.00). The tibial slope was decreased from (14.29±1.26) ° preoperatively to (9.31±0.79) ° postoperatively ( t=24.59, P=0.00) . The KT-1000 side-to-side difference decreased from (7.95±1.19) mm preoperatively to (1.79±0.49)mm postoperatively( t=18.34, P=0.00). At the last follow-up, Lysholm score, Hospital for Special Surgery score, Tegner score, and the IKDC knee evaluation score of patients showed significant improvement from preoperative( P<0.05). Conclusion:Arthroscopic combined with dual-plane high tibial osteotomy can get a good short term efficacy in the treatment of anterior cruciate ligament injury combined with varus deformity of knee joint which can significantly improve the alignment of lower extremity and knee joint stability.

Objective:To explore the short-term effectiveness of arthroscopic combined with dual-plane high tibial osteotomy in the treatment of anterior cruciate ligament injury combined with varus deformity of knee joint.Methods:A retrospective study was performed on 17 patients with anterior cruciate ligament injury combined with varus deformity of knee joint who underwent arthroscopic combined with dual-plane high tibial osteotomy at Department of Bone and Joint, the Affiliated Hospital of Southwest Medical University from January 2017 to June 2018.There were 11 males (11 knees) and 6 females (6 knees), aged 41.3 years (range: 32 to 49 years) .During the surgery, the weight bearing line of lower extremity was set to 62.5% position of the tibial plateau on coronal plane. The tibial slope was adjusted to the normal range on sagittal plane, and anterior cruciate ligament was reconstructed to improve the stability of knee joint.At final follow up, full length weight bearing X ray was used to evaluate the position of weight bearing line, femoral tibial angle and tibial slope pre- and post-operatively.The Lysholm scores, Hospital for Special Surgery score, Tegner knee activity scores and International Knee Documentation Committee (IKDC) scores were used to estimate knee joint function, while the Lachman test, KT-1000 side-to-side difference and pivot-shift test were used to estimate the knee joint stability.Results:The patients were followed up for 1.8 years(range:1.2 to 2.5 years). No complication such as infection, deep vein thrombosis, graft failure, nonunion or delayed union was observed.The weight bearing line was corrected from (28.48±2.24)% preoperatively to (57.43±1.02)% postoperatively ( t=46.80, P=0.00) .The femoral tibial angle was improved from (172.31±3.37) ° preoperatively to (178.91±1.34) ° postoperatively( t=10.46, P=0.00). The tibial slope was decreased from (14.29±1.26) ° preoperatively to (9.31±0.79) ° postoperatively ( t=24.59, P=0.00) . The KT-1000 side-to-side difference decreased from (7.95±1.19) mm preoperatively to (1.79±0.49)mm postoperatively( t=18.34, P=0.00). At the last follow-up, Lysholm score, Hospital for Special Surgery score, Tegner score, and the IKDC knee evaluation score of patients showed significant improvement from preoperative( P<0.05). Conclusion:Arthroscopic combined with dual-plane high tibial osteotomy can get a good short term efficacy in the treatment of anterior cruciate ligament injury combined with varus deformity of knee joint which can significantly improve the alignment of lower extremity and knee joint stability.

BACKGROUND: Integrin α5β1 has been shown to be related to acetabular cartilage degeneration due to developmental dysplasia of the hip.OBJECTIVE: To investigate the expression of integrin α5β1 in differently degenerated cartilage tissues of the knee after knee osteoarthritis.METHODS: Seventy-five degenerated articular cartilage specimens at the weight-bearing surface of femoral lateral and medial condyles were removed from patients with knee osteoarthritis who underwent total knee replacement, and were then allotted to four groups based on Mankin's grading system: normal (n=6), mild degeneration (n=24), moderate degeneration (n=26) and severe degeneration (n=19) groups. The expression of integrin α5β1 in each group was detected using real-time PCR, immunohistochemistry and western blot assay.RESULTS AND CONCLUSION: The detected results were consistent in the three kinds of assays, and the expression level of integrin α5β1 was ranked as follows: severe degeneration group > moderate degeneration group > mild degeneration group > normal group (P < 0.05). To conclude, differential expression levels of integrin α5β1 are found in different degenerative degrees of articular cartilage in knee osteoarthritis. Furthermore, the expression level of integrin α5β1 is increased with the degenerative severity.

Objective: To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function. Methods: Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS-PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr-activated SepharoseTM 4B. Reverse high pressure liquid chromatography (HPLC), SDS-PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone-antigen peptide complexes. Results: The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70-antigen peptide and 1.00 μg HSP90-antigen peptide activated lymphocytes significantly. Their A₄₉₀ values were 0.26±0.03, 0.45±0.05 and 0.32±0.04, respectively, while the corresponding doses of HSP60-antigen peptide and HSP110-antigen peptide did not activate lymphocytes. The killing rates of 1.00 μg HSP70-antigen peptide and 1.00 μg HSP70 were (45.0±2.0)% and (16.0±2.0)%, respectively, showing a significant difference (P=0.012). Similarly, the killing rates of 1.00 μg HSP90-antigen peptide and 1.00 μg HSP90 were (36.0±5.0)% and (13.0±4.0)%, respectively, also showing a significant difference (P=0.048). Conclusions The amount of chaperone antigen peptide complexes in gastric cancer cells is extremely low, but it is obviously increased in gastric cancer stem cells. After purification, the chaperone antigen peptide complexes with high purity can be prepared. The extracted chaperone antigen peptide complexes have stronger immunogenicity, and can be used to make tumor vaccine in vitro, which may have a good application value in the targeted therapy of gastric cancer.

There are multitudes of web resources that are quite useful for the microbial scientific research community. Here, we provide a brief introduction on some of the most notable microbial web resources and an evaluation of them based upon our own user experience.

Animals ; China ; Humans ; Immunophenotyping ; Influenza A Virus, H7N9 Subtype ; classification ; genetics ; immunology ; Influenza A virus ; classification ; genetics ; immunology ; Phylogeny