1.Clinical characteristics and virus identification of 4 cases of monkeypox confirmed in Zhejiang province
Lianqing LOU ; Xiaofei LI ; Zhicheng CHEN ; Linfang CHENG ; Linwei ZHU ; Xiaodi ZHANG ; Juncai TU ; Teng WANG ; Xiangcheng JIA ; Haiyan SHI ; Hangping YAO
Chinese Journal of Clinical Infectious Diseases 2023;16(4):256-261
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.
2.Preparation of azithromycin-ion exchange resin complexes for taste-masking and controlled drug release
Linfang TANG ; Yali YANG ; Shuwang HE ; Jing YAO
Journal of China Pharmaceutical University 2023;54(2):180-187
In order to mask the bitterness of azithromycin (AZI) and individually regulate the drug release rate to reduce gastrointestinal irritation, immediate-release AZI-AmberliteTM IRP64/HPC and delayed-release AZI-AmberliteTM IRP69/RS100 were prepared by modifying with hydroxypropyl cellulose (HPC) and Eudragit RS100, respectively, and further combined to achieve controlled release.The drug loading and drug utilization rate of AZI-ion exchange resin complexes were measured; the structure of AZI-ion exchange resin complexes was characterized by differential scanning calorimetry and X-ray diffraction; and the wetting humidity, odor masking effects, in vitro dissolution and release behaviors were determined.The results showed that the formation of AZI-ion exchange resin complexes changed the original crystallization state of the drug, that the 2.5% HPC-modified AZI-AmberliteTM IRP64/HPC and the 0.5% RS100-modified AZI-AmberliteTM IRP69/RS100 demonstrated good taste masking effect, and that their combination in the drug content ratio of 13∶67 achieved the expected drug release behavior, i.e.rapid release of AZI in the first 10 min and smooth release in the later 6 h.These results indicated that the AZI-ion exchange resin complexes prepared by surface modification and their composites could mask the bitterness of AZI and realize the flexible adjustment of drug release rate, which lays the foundation for the research and development of new AZI preparations.
3.Association of IFNL3/IFNL4 polymorphisms with efficacy of highly active antiretroviral therapy in patients with HIV-1 infection
Jianping YAO ; Dongshan YU ; Tianhao WENG ; Chenyu HU ; Linfang CHENG ; Tiansheng XIE ; Hangping YAO ; Limin FENG
Chinese Journal of Clinical Infectious Diseases 2018;11(1):28-35
Objective To investigate the associations of IFNL3/IFNL4 single-nucleotide polymorphisms(SNPs)with the efficacy of highly active antiretroviral therapy(HAART)in patients with HIV-1 infection.Methods Sixty-three adult patients with HIV-1 infection receiving HAART for at least 1 year in the First Affiliated Hospital, Zhejiang University School of Medicine were enrolled.HIV-1 RNA loads in plasma and HIV-1 DNA loads in peripheral blood mononuclear cells(PBMCs),and blood SNPs were detected by quantitative polymerase chain reaction(qPCR).Plasma inflammatory cytokines were examined by magnetic beads method,and the CD4 +T and CD8 +T lymphocyte counts in peripheral blood were measured by flow cytometry.According to response to HAART,the patients were classified as low HIV-1 RNA group(viral load <100 copies/mL)and high HIV-1 RNA group(viral load≥100 copies/mL);according to CD4+T lymphocyte counts,the patients were defined as low CD4+T cell group(<250 cells/μL), and high CD4+T cell group(≥250 cells/μL);according to HIV-1 DNA levels,the patients were divided into low(<100 copies/106cells)and high(≥100 copies/106cells)HIV-1 DNA groups.Results Three candidate SNPs rs368234815,rs8099917 and rs4803223 had significantly different distribution between low and high HIV-1 RNA groups(χ2=0.043,0.047 and 0.032,all P<0.05).The levels of interleukin(IL)-10 were declined in the low HIV-1 RNA group(U=4.00,P<0.05);the levels of IL-13 were decreased in the high HIV-1 RNA group and the high HIV-1 DNA group(U=0.00 and 2.00,both P<0.05);the levels of IL-21 were reduced in the high HIV-1 RNA group and in the low CD4 +T cell group(U=3.00 and 2.00, both P<0.05),the levels of IL-28A were decreased in the high HIV-1 RNA group,the high HIV-1 DNA group,and the low CD4 +T cell group(U=3.00, 0.50 and 3.00,P<0.05 or <0.01).In addition, rs368234815 was associated with IL-21 level(H=6.690,P<0.05),the IL-21 level in rs368234815 ΔG/ΔG [131.88(2.66,174.00)]was higher than that in TT/TT[6.79(2.81,26.48)](P<0.05);rs4803223 was correlated with IFN-γlevel(H=6.690, P<0.05),the IFN-γlevel in GG subtype[62.26(19.45, 96.49)]was higher than that in GA subtype[6.98(2.19, 99.14)](P<0.05).Conclusion The polymorphisms of IFNL3/IFNL4 rs368234815, rs8099917 and rs4803223 are associated with efficacy of HAART and immune-associated cytokines levels in patients with HIV-1 infection.
4.Correlation with neonatal congenital cytomegalovirus infection,maternal primary infection and secondary infection
Juying WANG ; Linfang YAO ; Feng LI
Chinese Journal of Immunology 2017;33(8):1205-1208
Objective:To explore the correlation with neonatal congenital cytomegalovirus infection,maternal primary infection and secondary infection.Methods: 48 neonates with congenital CMV infection were assigned to infection group with their mothers.And the other 30 couples without congenital CMV infection were assigned to negative group with their mothers.The level of CMV-IgM/IgG and affinity of CMV-IgG in peripheral blood were tested by CLIA,and CMV-DNA in mother′s milk,peripheral blood and urine of the newborn was tested by fluorescent quantitation PCR.We also analyzed the differences of the test results between the two groups and performed a retrospective analysis to compare the levels of CMV-IgG of the mother with early pregnancy with the result of this test.Results: In the infection group,the level of CMV-IgG in peripheral blood and CMV-DNA in milk was significantly higher than those in the negative group,the difference was statistically significant(P<0.01).Ratio of CMV-IgG antibody in newborn babies and their mothers in the infection group was lower than the other group,the difference was statistically significant(P<0.01).There was negative correlation of IgG level between the newborn babies and their mothers in the infection group,the difference was statistically significant(P<0.01);While in the positive correlation in the negative group,the difference was statistically significant(P<0.01).The CMV-IgG concentrations of the mothers with early pregnancy was significantly lower than that of this infection group,the difference was statistically significant(P<0.01);but in the negative group,there was no significant difference between the CMV-IgG level of the mothers with early pregnancy and the results of this test(P>0.05).Conclusion: It is a high-risk factor for neonatal congenital cytomegalovirus infection that CMV-IgG level of the pregnant women is promted by the reactivation or reinfection of cytomegalovirus.It is important to monitor CMV-IgM/IgG during pregnancy.
5.Advances in taste-masking technology of oral paediatric medicine
Linfang TANG ; Ziqiang ZHANG ; Rina SU ; Shuwang HE ; Jing YAO
Journal of China Pharmaceutical University 2017;48(2):135-141
As children are extremely sensitive to the bad taste of medicine,they have poor compliance with the bitter medicine.It is of great importance to develop the approaches of taste masking for the research of paediatric drug formulations.Besides,taste masking technology is one of the main barrier to develop children pharmaceutic preparation.This article provides an overview of the advance in taste masking technology of oral paediatric medicine in recent years,and introduces five types of taste masking technology in terms of drug,preparation and the bitter taste transduction,including principle and characteristics of these approaches,as well as their application in formulations,so as to provide some references for the development of paediatric medicine.
6.Diagnostic and therapeutical value of FPG,GA,HbA1c and GA/HbA1c ratio in T1 DM/T2 DM
Juying WANG ; Linfang YAO ; Lirong ZHOU ; Feng LI
Chinese Journal of Immunology 2015;(11):1536-1540
Objective:To investigate the diagnostic and terapeutical value of FPG,GA,HbA1c and GA/HbA1c ratio in T1DM/T2DM.Methods:The study was made by case-control method.In our study,30 healthy subjects were selected from health physical ex-amination as control group while 160 diabetics were selected as case group,in which there are 76 TIDM and 84 T2DM.Analyzing the difference of relevance of FPG,GA and HbA1c,the difference of GA/HbA1c and threshold of the case and the control,and this analysis was also used between the T1DM and the T2DM.The data was managed by independent-sample t test,ROCK and Pearson correlation test of SPSS.Results:The results of FPG,GA ,HbA1c and GA/HbA1c ratio of T1DM and T2DM were significantly higher than those in the control group(P<0.01).And they were higher in the T1DM than in the T2DM(P<0.01);in the T1DM group,GA was strongly positive correlative with HbA1c(P<0.01),FPG was weakly positive correlative with GA(P>0.05),and weakly negative correlative with HbA1c(P>0.05);in T2DM group,there were positive correlation among FPG,GA and HbA1c(P<0.05),the degree of correlation ranked as HbA1c/GA>FPG/GA>FPG/HbA1c;analyzing the ROC of measures in T1DM group,the sensitivity and specificity were re-spectively 86.8% and 100% for diagnosing DM when FPG threshold was set on 5.86 mmol/L ( AUC=0.922 ) ( P<0.05 ).The sensitivity and specificity were both 100% for detecting DM when GA threshold was set on 15.5%( AUC=1 ) ( P<0.05 ).The sensitivity and specificity were respectively 98.7%and 100%for diagnosing DM when HbA1c threshold was set to 6.10%( AUC=1) ( P<0.05).The sensitivity and specificity were respectively 93.4% and 100% for diagnosing DM when the threshold of GA/HbA1c was set on 2.95 ( AUC=0.992 ) ( P<0.05 );analyzing the ROC of measures in T2DM group, the sensitivity and specificity were respectively 91.7% and 100% for diagnosing DM when FPG threshold was set on 5.94 mmol/L ( AUC=0.941 ) ( P<0.05 ).The sensitivity and specificity were respectively 85.7%and 100%for diagnosing DM when GA threshold was set on 15.5%( AUC=0.977) ( P<0.05).The sensitivity and specificity were respectively 97.6% and 100% for diagnosing DM when HbA1c threshold was set on 5.95%(AUC=0.991)(P<0.05).The ratio of GA/HbA1c had no diagnostic cutoff point,AUC was 0.644(P>0.05).Conclusion:FPG,GA, HbA1c and GA/HbA1c ratio are of high value in monitoring of blood glucose, diagnosis and typing in T1DM and T2DM.There are missed diagnosis when we diagnose T1DM and T2DM by the upper limit of reagent instruestion of FPG,GA,HbA1c.It is more important for a person with T1DM to monitor FPG than others.
7.Ultrasound-guided percutaneous radiofrequency ablation treatment for renal clear cell carcinoma.
Xiangming CHENG ; Weidong GAN ; Linfeng XU ; Linfang YAO ; Feng QU ; Gutian ZHANG ; Xiaogong LI ; Hongqian GUO
Chinese Journal of Surgery 2014;52(11):856-860
OBJECTIVETo investigate clinical outcomes of ultrasound-guided percutaneous radiofrequency ablation (USG-RFA) in patients with renal clear cell carcinoma.
METHODSMedical records of 34 patients who underwent USG-RFA of renal clear cell carcinoma at the Department of Urology of the Affiliated Drum Tower Hospital of Medical School of Nanjing University from May 2009 to January 2014 were retrospectively reviewed, including 28 male and 6 female patients aged between 25 and 85 years (mean age 60.7 years). Of the included cases, 16 had tumors located in the left kidney, 16 in the right, 1 in the solitary kidney, and 1 in the bilateral kidney. There were 35 tumors in this study totally. The maximum diameter of the tumors was 1.8 to 5.0 cm (mean (2.7 ± 0.3) cm), of which 32 cases of renal tumors were ≤ 4.0 cm and 3 cases of renal tumors were > 4.0 cm to 5.0 cm. Pathological diagnosis were acquired by ultrasound-guided percutaneous biopsy after USG-RFA. Contrast-enhanced ultrasound was used to evaluate tumor outcomes at the time of the surgery, and multi-slice spiral CT enhanced scan and contrast-enhanced ultrasound were used to identify residues and recurrences after treatment.
RESULTSTreatments for all the patients were finished with short postoperative hospital stay about 3-5 days. No complications related to USG-RFA were encountered in any of the cases, such as perirenal fluid collection, perirenal hematoma, and peripheral organ damage. All the cases were diagnosed as clear cell carcinoma according to pathological results. The mean follow-up period was 29 ± 6 (range 3-59) months. Of the 35 USG-RFA-treated subjects, 32 tumors ≤ 4 cm reached the standard of complete treatment after one tumor was found with residue after the first month follow-up, and two tumors were noted recurrence at the 4 and 10 months follow-up after USG-RFA. Nonetheless, no residue or recurrence occurred after secondary treatment for these 3 tumors where pathological diagnosis were acquired again. The other 3 cases with tumors > 4.0 cm to 5.0 cm underwent USG-RFA twice or three times before reaching the standard of complete treatment, of which two had twice and one tumor had three times treatments. There was no carcinoma residue or recurrence during follow-up period.
CONCLUSIONSPercutaneous ultrasound-guided radiofrequency ablation for small renal mass (SRM) has satisfied clinical outcomes, with the advantage of less injury, lower complication rates and shorter recovery time for small size of renal clear cell carcinoma. USG-RFA may become the preferred treatment alternative for SRM.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Renal Cell ; surgery ; Catheter Ablation ; methods ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Retrospective Studies ; Treatment Outcome ; Ultrasonography, Interventional
8.IL-4 induced signal pathways regulating DC-SIGN expression in THP-1 cells
Linfang CHENG ; Changzhong JIN ; Lijuan WU ; Yijie WANG ; Juan WANG ; Hangping YAO ; Nanping WU
Chinese Journal of Microbiology and Immunology 2012;32(6):481-485
Objective To detect the signal pathways through which IL-4 regulates expression of DC-SIGN in THP-1 cells.Methods We used phorbol 12-myristate 13-acetate(PMA) differentiated THP-1 cells as the in vitro model of monocyte/macrophage cells to study the signal pathways involved in IL-4 regulated expression of DC-SIGN.DC-SIGN mRNA expression was detected by RT-PCR.Cytoplasmic DC-SIGN protein was tested by Western blot.Flow cytometry was used to detect cell surface expression of DC-SIGN.Cytoplasm and nuclear protein of PMA stimulated THP-1 cells induced by IL-4 for 0,10,20,30,60 and 120 min was extracted and detected by Western blot for signal pathway signaling protein and phosphoprotein.Results We found that a high expression of DC-SIGN could be induced by IL-4 at the levels of mRNA and cell surface protein.Up-regulated expression of DC-SIGN was almost completely blocked by the specific inhibitor of ERK pathway,and partly reduced by the specific inhibitors of JAK-STAT and NF-κB pathways.The activation of the three signaling pathways was directly confirmed by testing the phosphorylation of protein kinase within the cytoplasm and nucleus over time.Conclusion Multiple signaling pathways are involved in IL-4 induced high expression of DC-SIGN on THP-1 cells,in which ERK pathway is the main signal pathway.
9.Identification of gentianae macrophyllae radix using the ITS2 barcodes.
Kun LUO ; Pei MA ; Hui YAO ; Tianyi XIN ; Yan HU ; Sihao ZHENG ; Linfang HUANG ; Jun LIU ; Jingyuan SONG
Acta Pharmaceutica Sinica 2012;47(12):1710-7
DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.
10.Retroperitoneoscopic cryoablation for small renal tumors: a preliminary report
Huibo LIAN ; Hongqian GUO ; Weidong GAN ; Xiaogong LI ; Xiang YAN ; Shiwei ZHANG ; Tieshi LIU ; Feng QU ; Linfang YAO ; Gutian ZHANG
Chinese Journal of Urology 2010;31(6):369-372
Objective To present the technique and short-term results of retroperitoneal laparoscopic renal cryoablation for small renal tumors. Methods Ten selected patients cases with 11 renal tumors were included in present study. There were 3 cases of left renal tumor, 6 cases of right renal tumor and 1 case of bilateral renal tumors. Tumors were located at the upper pole (2), middle (6), or lower pole (3). All tumors were located distant from the collecting system, without evidence of metastatic disease. Mean tumor size was 2. 8 cm (range: 1.5-4.0). All the patients were managed with a double freeze-thaw cycle of retroperitoneal laparoscopic renal cryoablation. The preoperative Hb was (137± 21)g/L, ESR was (27±12)mm/1 h, SCr was (92±41)μmol/L, GFR was (42±10)ml/min.All the patients were taken routine biopsies. Results Cryoablation was technically successful in all 10 patients (11 tumors). The mean time of the operations was (101 ± 31) min, and the mean blood loss was (42±21) ml. None of the cases received blood transfusion post-operation. No operative complication was seen. The postoperative hospital stay was (4±2) d. The postoperative Hb was (129 ±18)g/L,ESR was (31±14)mm/1 h,SCr was (95±39)μmol/L,GFR was (40±11)ml/min. There was no statistic change of Hb, ESR, SCr and ECT-GFR after operations(P>0. 05). The biopsy results revealed that 8 tumors were renal clear cell carcinomas, and 2 tumors were papillary renal cell carcinomas, and 1 tumor was renal angiomyolipoma. All the patients had a minimum follow-up of 6 months (mean 16, range 6 to 21). Follow-up magnetic resonance imaging at 1, 3, and 6 months identified the punched-out, nonenhancing, spontaneously resorbing, renal cryolesions. Follow-up biopsie of the cryoablated tumor site was negative in the only patient who have undergone the biopsy. No evidence of local or port-site recurrence was found, and no metastatic disease. ConclusionsRetroperitoneal laparoscopic renal cryoablation for small renal tumors could be an accurate and effective intervention with a relatively low incidence of complications. Critical long-term data regarding laparoscopic renal cryoablation are awaited.

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