Effects of extramedullary disease on patients with newly diagnosed multiple myeloma / 中华血液学杂志

Objective: To summarize the characteristics of patients with newly diagnosed multiple myeloma (NDMM) admitted at Ruijin Hospital affiliated to Shanghai Jiaotong University School of Medicine. We compared the clinical characteristics and prognoses among patients with non-extramedullary disease (EMD), bone-related extramedullary (EM-B) disease, and extraosseous extramedullary (EM-E) disease and further explored the effects of autologous hematopoietic stem cell transplantation (ASCT) for EMD. Methods: From January 2015 to January 2022, data of 114 patients (22%) with EMD out of 515 patients with NDMM were retrospectively analyzed; 91 (18%) and 23 (4%) patients comprised the EM-B and EM-E groups, respectively. The clinical characteristics of patients in all groups were compared with the Chi-square test. Progression-free survival (PFS) and overall survival (OS) of patients were analyzed by the Kaplan-Meier method. Independent prognostic factors were determined using multivariate Cox proportional hazard model. Results: There were no significant differences in age, gender, ISS stage, light chain, creatinine clearance, cytogenetic risk, 17p deletion, ASCT, and induction regimens among the three groups. Overall, 13% of EM-E patients had IgD-type M protein, which was significantly higher than that in EM-B patients (P=0.021). The median PFS of patients in the non-EMD, EM-B, and EM-E groups was 27.4, 23.1, and 14.0 months; the median OS was not reached, 76.8 months, and 25.6 months, respectively. The PFS (vs non-EMD, P=0.004; vs EM-B, P=0.036) and OS (vs non-EMD, P<0.001; vs EM-B, P=0.002) were significantly worse in patients with EM-E, while those were not significantly different between patients with EM-B and those with non-EMD. In the multivariate analysis, EM-E was an independent prognostic factor for OS in patients with NDMM (HR=8.779, P<0.001) and negatively impacted PFS (HR=1.874, P=0.050). In those who did not undergo ASCT, patients with EM-B had significantly worse OS than those with non-EMD (median 76.8 months vs. not reached, P=0.029). However, no significant difference was observed in the PFS and OS of patients with EM-B and those with non-EMD who underwent ASCT. Conclusions: Compared to patients with either non-EMD or EM-B, those with EM-E had the worst prognosis. EM-E was an independent risk factor for OS in patients with NDMM. ASCT can overcome the poor prognosis of EM-B.

Newly developed gas-assisted sonodynamic therapy in cancer treatment

Sonodynamic therapy (SDT) is an emerging noninvasive treatment modality that utilizes low-frequency and low-intensity ultrasound (US) to trigger sensitizers to kill tumor cells with reactive oxygen species (ROS). Although SDT has attracted much attention for its properties including high tumor specificity and deep tissue penetration, its anticancer efficacy is still far from satisfactory. As a result, new strategies such as gas-assisted therapy have been proposed to further promote the effectiveness of SDT. In this review, the mechanisms of SDT and gas-assisted SDT are first summarized. Then, the applications of gas-assisted SDT for cancer therapy are introduced and categorized by gas types. Next, therapeutic systems for SDT that can realize real-time imaging are further presented. Finally, the challenges and perspectives of gas-assisted SDT for future clinical applications are discussed.

Effect and safety of aflibercept in the treatment of polypoidal choroidal vasculopathy with ranibizumab-resistant serous pigment epithelial detachment / 中华实验眼科杂志

Objective:To evaluate the effectiveness and safety of intravitreal injection of different doses of aflibercept for polypoidal choroidal vasculopathy (PCV) with serous pigment epithelial detachment (PED) resistant to ranibizumab.Methods:A non-randomized controlled clinical study was conducted.Seventy-three eyes of 73 patients with PCV and serous PED resistant to ranibizumab were enrolled at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2020.All patients were treated by intravitreal injection of 2 mg or 4 mg aflibercept according to patients' willingness.2 mg aflibercept or 4 mg aflibercept was intravitreally injected monthly for three consecutive months following pro re nata (PRN) regimen in 2 mg aflibercept group (38 eyes) and 4 mg aflibercept group (35 eyes), respectively.PED height and central macular thickness (CMT) were measured by optical coherence tomography, and the best corrected visual acuity (BCVA) was examined with a visual acuity chart and converted to logarithm of the minimum angle of resolution (LogMAR) unit before injection and 1 month, 2, 3, 6 months from the first injection.Intraocular pressure and treatment-related adverse events were recorded.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2021-KY-1252).Written informed consent was obtained from each patient prior to entering study cohort.Results:Thirty-three patients (86.84%) in 2 mg aflibercept group and 30 patients (85.71%) in 4 mg aflibercept group finished the treatment and follow-up, respectively. The PED, BCVA and CMT before treatment and at the end of follow-up were (379.24±95.50) and (280.09±120.50)μm, 0.68±0.27 and 0.51±0.19, (393.96±100.81) and (291.70±44.09)μm in 2 mg aflibercept group, respectively, showing statistically significant differences (all at P<0.05).The PED, BCVA and CMT before treatment and at the end of follow-up were (393.07±93.76) and (278.63±145.07)μm, 0.66±0.31 and 0.48±0.22, (377.43±79.61) and (284.67±84.88)μm in 4 mg aflibercept group, respectively, with statistically significant differences (all at P<0.05).The CMT value in 4 mg aflibercept group was significantly lower than that in the 2 mg aflibercept group in one month after injection ( P<0.05).No severe ocular and systemic adverse events were found during the follow-up, such as retinal detachment, endophthalmitis, cataract, and persistent high intraocular pressure. Conclusions:Both 2 mg and 4 mg aflibercept can effectively treat ranibizumab-resistant PCV with serous PED, and improve the anatomical structure of retina and BCVA.4 mg aflibercept can accelerate the recovery of PED and CMT.

Analysis of BEST1 gene mutation and clinical phenotype in two families with Best vitelliform macular dystrophy and autosomal recessive bestrophinopathy / 中华眼底病杂志

Objective:To report the BEST1 gene mutations and clinical phenotypes in two pedigrees with Best vitelliform macular dystrophy (BVMD) and autosomal recessive bestrophinopathy (ARB). Methods:A retrospective clinical study. From November 2019 to March 2021, in the Department of Ophthalmology of The First Affiliated Hospital of Zhengzhou University, the BVMD family (4 patients and 6 family members) and the ARB family (2 patients, 2 family members), a total of 6 patients and 8 normal family members were included in the study. Detailed medical history was obtained; best corrected visual acuity, fundus color photography, electrophysiology, optical coherence tomography and fundus autofluorescence examination were performed. The clinical characteristics for all patients in the two families were analyzed. Three milliliter peripheral venous blood of all participants in the family was collected, and the whole genomic DNA was extracted with gene sequencing using next-generation sequencing technology based on targeted capture. Compared with the database to identify the pathogenicity mutation sites, suspected pathogenic mutation sites were selected, then mutations in other members in the family was assayed by Sanger sequencing.Results:In family 1, the proband was demonstrated as typical BVMD, other patients were multifocal vitelliform macular dystrophy. The DNA sequencing result showed that all the 4 patients carried heterozygous missense mutations in exon 3 of BEST1 gene: c.240C>G (p.F80L) (M1) and 2 members carried this mutation, but without clinical phenotype. M1 was a likely-pathogenic mutation reported for the first time. In family 2, the proband and the other patient were diagnosed as ARB. The DNA result showed that the 2 patients carried heterozygous missense mutations in exon 5 and exon 2 of BEST1 gene: c.584C>T (p.A195V) (M2)、c.139C>A (p.R47S) (M3), and a heterozygous frameshift mutation in exon 3 of BEST1 gene: c.235dupT (p.S79Ffs*153) (M4). M2 was a pathogenic mutation reported previously. M3 variant was of undetermined significance. M4 was a first reported pathogenic mutation. Conclusions:The BEST1 gene mutation is the main cause of BVMD and ARB. Different mutation sites have different clinical phenotypes. BVMD and ARB have genetic and clinical heterogeneity.

Predictive value of MB isoenzyme of creatine kinase and poisoning severity score in the prognosis of patients with wasp sting / 中华危重病急救医学

Objective:To explore the predictive value of MB isoenzyme of creatine kinase (CK-MB) and poisoning severity score (PSS) in the clinical prognosis of patients with wasp sting.Methods:A retrospective study was conducted. The clinical data of patients who were stung by wasps admitted to emergency department of the First Affiliated Hospital of Henan University of Science and Technology from July 2017 to November 2019 were collected. The 24-hour acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), CK-MB and PSS scores of the patients were collected after admission, and 28-day outcome was recorded. Spearman correlation analysis method was used to analyze the correlation between CK-MB and PSS score. Logistic regression model was used to construct joint predictors, and the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of various indicators for 28-day prognosis of patients with wasp stings.Results:Finally 90 patients were included in the analysis. There were 67 patients survived at 28 days, and 23 dead with the 28-day mortality of 25.6%. APACHEⅡ score, CK-MB and PSS score in the death group were significantly higher than those in the survival group [APACHEⅡscore: 19.7±2.7 vs. 13.7±2.3, CK-MB (U/L): 183 (151, 243) vs. 36 (21, 75), PSS score: 17.7±2.6 vs. 9.3±4.5, all P < 0.01]. The correlation analysis showed that CK-MB and PSS score were positively correlated ( r = 0.843, P < 0.01). Logistic regression model fitted CK-MB and PSS score, and Hosmer-Lemeshow test showed that the model fitted well. ROC curve analysis showed that the area under ROC curve (AUC) of CK-MB for predicting 28-day outcome was 0.957, the sensitivity was 91.3%, and the specificity was 88.1%; the AUC of PSS score was 0.908, the sensitivity was 91.3%, and the specificity was 90.8%. The AUC of CK-MB combined with PSS score was 0.964, the sensitivity was 100%, and the specificity was 79.4%, indicating that CK-MB combined with PSS score had higher predictive value and higher sensitivity for 28-day prognosis of patients with wasp sting. Conclusions:High CK-MB level and high PSS score in early stage of wasp sting injury indicate poor prognosis. Both CK-MB and PSS score can be used as predictors for predicting the prognosis of patients with wasp stings. In addition, CK-MB combined with PSS score have greater predictive value .

Local infiltration of tissue-resident memory CD4 + T cells in lesions of patients with pemphigus and its clinical implications / 中华皮肤科杂志

Objective:To investigate the local infiltration of tissue-resident memory CD4 + T (CD4 + T RM) cells in lesions of patients with pemphigus and its clinical implications. Methods:From September 2017 to December 2018, 20 patients with pemphigus and 15 healthy human controls were collected from Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. Flow cytometry was performed to determine the proportion of CD4 + T RM cells in skin lesions of pemphigus patients and normal skin of healthy controls. The degree of CD4 + T RM cell infiltration in skin lesions was compared among different body sites of the patients with pemphigus, and the correlation between the proportion of CD4 + T RM cells and the time to disease control was analyzed. Normally distributed data were analyzed by using t test, and non-normally distributed data by using non-parametric test; the Pearson correlation coefficient was used to analyze correlations of the proportion of CD4 + T RM cells with pemphigus disease area index (PDAI) scores and circulating anti-desmoglein (Dsg) antibody titers. Results:Among the 20 patients, there were 16 with pemphigus vulgaris and 4 with pemphigus foliaceus. All the patients had skin involvement, 14 lesional tissue samples were taken from the trunk, and 6 from the limbs. There was no significant difference between the healthy control group and pemphigus group in terms of age, gender or biopsy sites (all P > 0.05) . The proportions of CD3 + T cells (72.75% ± 8.22%) and CD4 + T RM cells (44.05% ± 14.27%) in the skin lesions of patients with pemphigus were significantly higher than those in the skin tissues of the healthy controls (31.33% ± 8.72%, 12.60% ± 5.12%, t = 14.24, 9.10, respectively, both P < 0.001) . Among the patients with pemphigus, the proportion of CD4 + T RM cells was significantly higher in the skin lesions on the trunk (49.57% ± 12.32%) than in those on the limbs (31.17% ± 9.75%, t = 3.23, P < 0.05) . The proportion of CD4 + T RM cells in the skin lesions was positively correlated with the PDAI scores ( r2 = 0.246, P = 0.026) , but not correlated with serum titers of circulating anti-Dsg1 ( r2 = 0.137, P > 0.05) or anti-Dsg3 ( r2 = 0.162, P > 0.05) antibodies in the patients. During the treatment with systemic glucocorticoids, the proportion of CD4 + T RM cells in the skin lesions was significantly higher in the patients whose lesions could not be controlled within 4 weeks than in those whose lesions could be controlled within 4 weeks ( t = 3.22, P < 0.05) . Conclusion:The proportion of CD4 + T RM cells markedly increased in the skin lesions of patients with pemphigus, which may be related to the severity of the disease and response to treatment.

Diagnostic accuracy of recognition of stroke in the emergency room scale: a Meta-analysis / 中华危重病急救医学

Objective To systematically evaluate the diagnostic accuracy and clinical applicability of recognition of stroke in the emergency room (ROSIER) scale by systematic review and Meta-analysis. Methods The Chinese and English literatures concerning the diagnostic accuracy of ROSIER published from January 1st 2005 to December 31st 2018 by PubMed, Embase, Wanfang, VIP and CNKI databases were searched comprehensively and systematically. The sensitivity, specificity, and diagnostic odds ratio (DOR) of ROSIER in total population and subgroup analysis were pooled by using bivariate mixed effects model. Sensitivity analysis was used to evaluate the stability of the results. Deek funnel plot was utilized to evaluate publication bias. The clinical applicability of ROSIER was evaluated by Fagan Nomogram. Results A total of 28 studies incorporating 7 579 subjects were enrolled in this Meta-analysis. Meta-analysis in total population showed that the pooled sensitivity, specificity and DOR of ROSIER was 0.89 [95% confidence interval (95%CI) = 0.86-0.91, P = 0.00], 0.74 (95%CI = 0.67-0.80, P = 0.00) and 22.09 (95%CI =14.86-32.82, P = 0.00), respectively. Subgroup analysis of pooled sensitivity of ROSIER showed that Asian patients was significantly higher than European patients [0.89 (95%CI = 0.86-0.92) vs. 0.74 (95%CI = 0.66-0.82), P < 0.01], prospective study was significantly higher than retrospective study [0.89 (95%CI = 0.87-0.92) vs. 0.74 (95%CI = 0.61-0.88), P < 0.05], pre-hospital emergency was significantly higher than emergency department [0.87 (95%CI = 0.80-0.94) vs. 0.85 (95%CI = 0.81-0.90), P < 0.01], study with sample size ≤ 200 was significantly higher than study with sample size > 200 [0.88 (95%CI = 0.83-0.93) vs. 0.82 (95%CI = 0.76-0.88), P < 0.05], but there was no significant difference between different evaluators or different male to female ratio subgroups. Subgroup analysis of pooled specificity of ROSIER showed that European patients was significantly higher than Asian patients [0.81 (95%CI = 0.73-0.89) vs. 0.79 (95%CI = 0.73-0.85), P < 0.05], retrospective study was significantly higher than prospective study [0.88 (95%CI =0.78-0.97) vs. 0.79 (95%CI = 0.73-0.84), P < 0.05], pre-hospital emergency was significantly higher than emergency department [0.82 (95%CI = 0.73-0.91) vs. 0.79 (95%CI = 0.73-0.85), P < 0.01], emergency physicians was significantly higher than other medical workers [0.80 (95%CI = 0.74-0.86) vs. 0.79 (95%CI = 0.69-0.90), P < 0.05], study with sample size ≤ 200 was significantly higher than study with sample size > 200 [0.82 (95%CI = 0.76-0.89) vs. 0.78 (95%CI = 0.71-0.85), P < 0.05], but there was no significant difference between different male or female ratio subgroups. Sensitivity analysis showed that there was no significant change in pooled DOR before and after excluding each study, indicating that the results were stable. Funnel plot showed that there was a significant publication bias in the total population (P = 0.04), but there was no publication bias in the European population (P = 0.57) or the Asian population (P = 0.08). According to the results of the Fagan Nomogram, with the pretest probability of 50%, when ROSIER was positive, the probability of being diagnosed with stroke increased to 77%, and when ROSIER was negative, the probability of being diagnosed with non-stroke decreased to 13%. It was suggested that ROSIER had good applicability and high clinical diagnostic value. Conclusions ROSIER has high diagnostic sensitivity and specificity, and has high clinical diagnostic value. It is a valid stroke identification tool which can be widely used in Asian population, pre-hospital emergency and be utilized by trained medical worker.

Analysis of treatments for 86 cases type Ⅲa hepatolithiasis without hepatectomy / 中国实用外科杂志

OBJECTIVE: To explore therapeutic strategies of type Ⅲa hepatolithiasis. METHODS: The clinical data of 86 patients with type Ⅲ a hepatolithiasis admitted from July 2014 to December 2017 in Department of Hepatobiliary Surgery, the First Affiliated Hospital of Guangxi University of Chinese Medicine were retrospectively analyzed. Patients without liver resection were included. Variables including residual stones,times of choledochoscopy performing,stone clearance rate and recurrence rate were observed. RESULTS: There was a total of 23 patients with biliary strictures and 2 cases of bile leakage after operation,83 patients(96.5%) residual stones after Initial treatment. About six weeks after surgery,the choledochoscopy was performed via T tube sinus or subcutaneous blind loop. The median lithotomy times was 3. There were 4 cases of residual small stones,and stone clearance rate was 95.4%. Median follow-up time was 24 months. 6 patients(7.0%) had recurrent intrahepatic bile duct stones,and the proficiency was 93.0%(80/86). 6 patients(7.0%) had sporadic cholangitis. CONCLUSION: It is safe and feasible to correct the stricture and remove the stones through the combination of biliary surgery and choledochoscope for type Ⅲa hepatolithiasis. The short-term outcomes are satisfactory.

Role of eosinophils in the pathogenesis of bullous pemphigoid / 中华皮肤科杂志

Bullous pemphigoid (BP) is the most common autoimmune subepidermal blistering skin disease,and a large number of eosinophils (EOS) obviously infiltrate the dermis of BP lesions.Additionally,EOS and their activated cytokines and chemokines are abundantly present in blisters and peripheral blood of BP patients.It is also proved that EOS can induce the dermal-epidermal separation.The level of interleukin (IL)-5 is markedly increased in the sera and blister fluids of BP patients,and IL-5 secreted by Th2 cells and EOS can regulate the differentiation,activation and survival of EOS.All the above evidence indicates that EOS are associated with the occurrence of BP.Researches on EOS and their cytokines will develop a new field of targeted therapy for BP.

Impacts of multicomponent environment on intrinsic dissolution rate of berberine in biopharmaceutics classification system of Chinese materia medica / 中国中药杂志

To illustrate the intrinsic dissolution rate(IDR) involved in biopharmaceutics classification system of Chinese materia medica(CMMBCS), investigate the effect of artificial multicomponent environment on IDR of berberine, by using a non-disintegrating disk, and summarize related rules by using the obtained data. Progressive levels design was used to investigate the effects of single component environment (puerarin, baicalin, and glycyrrhizic acid); double-component environment(puerarin+baicalin, puerarin+ glycyrrhizic acid, baicalin+glycyrrhizic acid); and triple-component environment(puerarin+baicalin+glycyrrhizic acid) on IDR of berberine, laying a foundation for further researches on the absorption mechanism of multiple components.

Pathological features of ectopic lymphoid structure in skin lesions of patients with pemphigus / 中华皮肤科杂志

Objective To investigate pathological features of infiltrating lymphocytes in skin lesions of patients with pemphigus,and to analyze their correlation with titers of anti-desmoglein (Dsg) 1 and anti-Dsg3 antibodies in peripheral blood.Methods A retrospective pathological analysis was performed in 93 patients with pemphigus vulgaris or pemphigus foliaceus,who visited the Department of Dermatology of Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine between 2014 and 2016.For each HE-stained section,the total number of lymphocytes per × 50 microscopic field was counted,and defined as lymphocyte density index.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in the patients with pemphigus.The correlations between the lymphocyte density index and titers of anti-Dsg1 and anti-Dsg3 antibodies were analyzed.Immunohistochemical staining was performed in lesional skin samples from 8 patients with pemphigus vulgaris and 8 patients with pemphigus foliaceus,so as to analyze the distribution of CD3+ T cells,CD20+ B cells and CD138+ plasma cells.Results Of the 93 pathological sections,93 (100.00％) showed Grade1 lymphocyte aggregates,64 (68.09％) showed Grade 2 lymphocyte aggregates,and 10 (10.64％) showed Grade 3 lymphocyte aggregates,and the 56 cases of pemphigus vulgaris and 37 of pemphigus foliaceus showed the similar proportion of grade 1,2 and 3 lymphocyte aggregates.There was also no significant difference in the lymphocyte density index between patients with pemphigus vulgaris and pemphigus foliaceus (P ＞ 0.05),and the lymphocyte density index was uncorrelated with the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in patients with pemphigus.Of the 16 cases of pemphigus,CD3+ T cells were found in all cases,CD20+ B cells in 15,and CD138+ plasma cells in 12.Of 16 sections,all showed a large amount of CD3+ T cells in Grade 1-3 lymphocyte aggregates,while lymphocyte aggregates containing CD20+ B cells and CD138+ plasma cells were found in 52.80％ ± 5.78％ and 34.59％ ± 7.42％ of sections respectively.No significant differences in the distribution of CD3+ T cells,CD20+ B cells,CD138+ plasma cells were found between the 8 cases of pemphigus vulgaris and 8 cases of pemphigus foliaceus.Conclusion Different degrees of lymphocyte infiltration generally exist in skin lesions of patients with pemphigus,which may form ectopic lymphoid structures and contribute to the development and aggravation of pemphigus skin lesions.

Analysis of Genetic Polymorphism and Genetic Distance of 19 Autosomal STR Loci in Jiangsu Han Population / 法医学杂志

Objective To investigate the distribution of alleles in 19 autosomal short tandem repeat (STR) loci in Jiangsu Han population.Methods Goldeneye20A kit was used to detect 9 025 samples.Genetic analysis was performed on typing data of 19 autosomal STR loci, and genetic distance with other 17 populations was analyzed.Results All the 19 autosomal STR loci were consistent with the Hardy-Weinberg equilibrium (P＞0.05), with the heterozygosity 0.616 1-0.916 3, probability of match 0.012 8-0.202 6, discrimination power 0.797 4-0.987 2, probability of paternity exclusion 0.310 8-0.828 8, and polymorphic information content 0.561 7-0.913 6.The cumulative discrimination power and cumulative probability of exclusion were 0.999 999 999 999 999 998 434 1 and 0.999 999 989, respectively.The Jiangsu Han population had close genetic distances with the Han population in Tianjin, Hunan and Jilin, and significant difference with Han population in Aletai region in Xinjiang (P＜0.05).Conclusion The STR allele polymorphism data and population genetic parameters of Jiangsu Han population can provide data support for the forensic application of these STR loci in Jiangsu Han population.

Standardization of preparation of rice fried Codonopsis Radix based on correlation analysis of functional component and shape index / 中国中药杂志

The quality standard of Chinese herbal medicine is the key in quality control, and the establishment of rice processing technology standard of rice fried Codonopsis Radix has important significance for the overall quality control. In this study, the color brightness (L*), red-green chromaticity coordinate (a*), yellow-blue chromaticity coordinate (b*) and total color value（E*ab）were determined by colorimeter and used as the shape indexes. Ethanol extractives, polysaccharide, lobetyolin and 5-hydroxymethyl furfural contents were determined by using high performance liquid chromatography (HPLC) and ultraviolet (UV), and used as chemical indexes in the analysis by normalization distance method. The two index methods were adopted to optimize the processing technology of rice fried Codonopsis Radix by taking processing temperature, processing time, rice types and rice dosage as factors. The correlation analysis between the two index methods was carried out by SAS 9.40 and JMP 11 software. The results showed that all the four factors had significant differences, and the best processing technology was as follows: polished round-grained rice as raw material, processing temperature at 140 °C, processing time of 10min, and the ratio of Codonopsis pilosula and rice is 100:30. Chemical indexes showed significant positive correlations with shape indexes, providing basis for the quality evaluation of rice fried Codonopsis Radix in processing.

Expression and specific antibody-producing capacity of B lymphocytes infiltrating in lesions of patients with pemphigus vulgaris / 中华皮肤科杂志

Objective To evaluate the specific antibody-producing capacity of locally infiltrating B lymphocytes in lesions of patients with pemphigus vulgaris (PV).Methods Totally,35 patients with PV and 22 healthy controls were enrolled into this study.Skin tissues were resected from blisters or erosions of the patients with PV,and from normal skin of healthy controls.Then,mononuclear cells were isolated from these skin tissues.Flow cytometry was performed to determine the percentages of lymphocytes,CD 19+ B lymphocytes,and desmoglein (Dsg)1-and Dsg3-specific CD19+ B lymphocytes.B lymphocytes isolated from the lesional skin of patients with PV were cultured in vitro.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supernatant.Receiver operating characteristic (ROC) curve analysis was conducted to calculate positive rates of anti-Dsg1 and anti-Dsg3 antibodies.Results The percentages of lymphocytes (17.95％ ± 3.85％) and CD19+ B lymphocytes (4.27％ ± 1.13％) were significantly higher in the lesional skin of PV patients than in the normal skin of healthy controls (7.83％ ± 1.29％,0.61％ ± 0.31％ respectively;t =2.49,U =13.00 respectively,both P ＜ 0.05).Among the CD19+ B lymphocytes in the lesional skin of PV patients,the percentage of CD19qgG+ B cells was (38.33 ± 5.56)％,and percentages of Dsg1-and Dsg3-specific CD19+ B lymphocytes were 12.87％ ± 1.267％ and 10.42％ ± 1.243％ respectively.After the in vitro culture for 6 days,the titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supematant were (4.89 ± 1.56) U/ml and (35.45 ± 13.03) U/ml respectively,with their positive rates being 85％ (17/20)and 95％ (19/20) respectively.Conclusion There are Dsg1-and Dsg3-specific B lymphocytes aggregating in the lesional skin of patients with PV,which can produce anti-Dsg1 and anti-Dsg3 antibodies after in vitro culture.

Genetic Polymorphism of 30 InDel Loci in Han Population from Jiangsu Province / 法医学杂志

Objective To investigate the genetic information of 30 insertion/deletion (InDel) loci in Han population from Jiangsu Province, and to explore the application values of Investigator? DIPplex kit for guiding the forensic analysis in Han population from Jiangsu Province. Methods The autosomal InDel loci of 305 unrelated healthy Han individuals from Jiangsu Province were genotyped and analysed by In-vestigator? DIPplex kit, and the allelic frequencies and forensic parameters of 30 InDel loci were statis-tically analysed. Results The distribution of 30 InDel loci in Han population from Jiangsu Province con-formed to Hardy-Weinberg equilibrium. The minor allele frequencies of 21 InDel loci were above 0.3. The polymorphism information content ranged from 0.089 to 0.375, while the discrimination power dis-tributed from 0.093 to 0.500. The paternity exclusion in duo cases and trio cases were 0.047-0.250 and 0.046-0.219, respectively. The linkage disequilibrium analysis of 30 InDel loci showed that all loci were independent from each other. The combined discrimination power was 1-7.369 ×10-8, whereas the com-bined mean exclusion chance in duo cases was 0.998933978, in trio cases was 0.997806392. The Fst values were all less than 0.06 except HLD118 and other four loci, which showed small differences be-tween groups. Conclusion The InDel loci of Investigator ? DIPplex kit can be used as complementary genetic markers for the cases associated with forensic genetics.

Overall intestinal permeability of multiple components in lotus leaf by in situ single pass intestinal perfusion models / 中国中药杂志

To investigate the overall intestinal permeability of multiple components in lotus leaves and make clear the interaction in composition absorption process. Rat single-pass intestinal perfusion technique was used, and the results showed that the Peff values of nuciferine, demethylanuciferine, rutin, quercetin, kaempferol from lotus leaf were greater than 0.5×10⁻⁴ cm•s⁻¹. In the biopharmaceutics classification system (BCS) intestinal permeability property, these ingredients were high permeable components, while the hyperin was low permeable component. However, in the multi-component environment of the lotus leaf extract, component permeation was changed. Semi quantitative analysis of the unclear components showed that under the multi-component environment, four in seven components with relatively high contents had a Peff value less than 0.5×10⁻⁴ cm•s⁻¹, indicating these 4 components were of low permeability, while other 3 components were of high permeability. The results could be valuable to make clear the overall intestinal permeability of multiple components in lotus leaf, and lay a foundation for studying the mechanism of the lipid-lowering effect of lotus leaf.

Validation of in situ single pass perfusion model based on P-gp / 中国中药杂志

To validate in situ rats intestinal single pass perfusion model based on P-glycoprotein (P-gp). Firstly, phenol red perfusion was carried out to verify the close connection structure of intestinal epithelial cells, and the integrity of the intestinal epithelium, with a gravimetric method for correcting water flux. The level of phenol red was determined by high performance liquid chromatography (HPLC) both before and after perfusion. Secondly, the positive drug digoxin specified by FDA was used to validate the model. After different mass concentrations of verapamil were given in the rats, the absorption parameters of digoxin in ileum of rats were observed and compared. The results showed that the phenol red was absorbed in rats ileum segment, with an effective permeability coefficient of (1.09±0.62)×10 ⁻⁶ cm•s ⁻¹. The experiment results indicated that the close connection structure of intestinal epithelial cells was normal, and the integrity of the intestinal epithelium was maintained well. In digoxin perfusion experiment, in case no verapamil was given, digoxin showed certain degree of absorption in rat ileum, with an effective permeability coefficient (Peff) of (1.07±0.59)×10 ⁻⁵ cm•s ⁻¹; after mass concentrations of 0.01,0.1 mmol•L ⁻¹ verapamil were given, the absorption of digoxin was on the rise in rat ileum, with an effective permeability coefficient Peff of (1.58±0.69)×10 ⁻⁵, (3.28±0.95)×10 ⁻⁵ cm•s ⁻¹ respectively (P<0.05). Digoxin perfusion experiment verified that P-gp expression in small intestine epithelium was intact and can be used in the research of P-gp efflux transporter.

In vitro and in vivo intestinal absorption of Huanglian decoction in multi-component environment / 中国中药杂志

The effect of drugs in the body is always inseparable with the dynamic processes of absorption, distribution, metabolism and excretion. For oral drug, absorption, as the first link to enter the body, is particularly valued. Traditional Chinese medicine has multiple components and multiple targets, and the nature of its single composition is different from that in the multicomponent environment. Alkaloids in Huanglian extract was used as the main object in this study to establish an analytical method for determining the content of alkaloids in Huanglian. In addition, the compositions of the Huanglian aqueous extract solution which can be absorbed through intestinal wall into blood, were initially determined by the means of everted gut sac and in intestinal perfusion with venous sampling experiment. This paper can provide data reference and support for the further study on the absorption and metabolism of Huanglian.

Study on biopharmaceutics classification system for Chinese materia medica of extract of Huanglian / 中国中药杂志

One of the advantages of biopharmaceutics classification system of Chinese materia medica (CMMBCS) is expanding the classification research level from single ingredient to multi-components of Chinese herb, and from multi-components research to holistic research of the Chinese materia medica. In present paper, the alkaloids of extract of huanglian were chosen as the main research object to explore their change rules in solubility and intestinal permeability of single-component and multi-components, and to determine the biopharmaceutical classification of extract of Huanglian from holistic level. The typical shake-flask method and HPLC were used to detect the solubility of single ingredient of alkaloids from extract of huanglian. The quantitative research of alkaloids in intestinal absorption was measured in single-pass intestinal perfusion experiment while permeability coefficient of extract of huanglian was calculated by self-defined weight coefficient method.

Discussion on research thinking of traditional Chinese medicine standardization system based on whole process quality control / 中国中药杂志

The concept of "Quality by design" indicates that good design for the whole life cycle of pharmaceutical production enables the drug to meet the expected quality requirements. Aiming at the existing problems of the traditional Chinese medicine (TCM) industry, the TCM standardization system was put forward in this paper from the national strategic level, under the guidance by the idea of quality control in international manufacturing industry and with considerations of TCM industry's own characteristics and development status. The connotation of this strategy was to establish five interrelated systems: multi-indicators system based on tri-indicators system, quality standard and specification system of TCM herbal materials and decoction pieces, quality traceability system, data monitoring system based on whole-process quality control, and whole-process quality management system of TCM, and achieve the whole process systematic and scientific study in TCM industry through "top-level design-implement in steps-system integration" workflow. This article analyzed the correlation between the quality standards of all links, established standard operating procedures of each link and whole process, and constructed a high standard overall quality management system for TCM industry chains, in order to provide a demonstration for the establishment of TCM whole-process quality control system and provide systematic reference and basis for standardization strategy in TCM industry.