Optimization of Agrobacterium-Mediated Genetic Transformation System of Armillaria gallica / 中国实验方剂学杂志

ObjectiveTo optimize the existing genetic transformation system of Armillaria gallica to improve the transformation efficiency and lay a foundation for the follow-up research on Armillaria molecular marker-assisted breeding and gene function. MethodThe genetically transformed plasmid pH101-PAgGPD-GFP-TrpC was constructed，transformed into Escherichia coli，amplified， and cultured，and the plasmid was extracted. The extracted plasmid was transformed into four different agrobacteria LBA4404，EHA105，GV3101，and AGL-1，respectively. The transformed agrobacteria were used for impregnating A. gallica，and the agrobacteria with the highest conversion rate were screened out. Then the agrobacterium-mediated genetic transformation system of A. gallica was optimized from the type and concentration of antibiotics，co-culture time，concentration of bacterial solution， and impregnation method. The phenotype profiles of A. gallica under different conditions were observed using Synbiosis ProtoCol 3. ResultThe optimized genetic transformation conditions of A. gallica were as follows： the Agrobacterium strain of EHA105 at absorbance A600 nm=0.6， the co-culture time of 2 d， the infection mode of negative pressure impregnation for 10 min， the primary screening medium of PDA medium containing 400 mg·L-1 cefotaxime sodium and 10 mg·L-1 hygromycin，and the secondary screening medium of PDA medium containing 12 mg·L-1 hygromycin. ConclusionIn this study，the existing genetic transformation system of A. gallica was optimized，and there was a significant difference in the transformation rate before and after optimization （P<0.05）. After optimization，the transformation efficiency of A. gallica was about 4.33%，which was about eight times higher than that before optimization.

Efficacy and safety of lenvatinib combined with sintilimab as the second-line therapy for intrahepatic cholangiocarcinoma / 临床肝胆病杂志

Objective To investigate the efficacy and safety of lenvatinib combined with sintilimab as the second-line therapy for advanced intrahepatic cholangiocarcinoma (ICC). Methods A retrospective analysis was performed for the clinical data of the patients with advanced ICC who were admitted to Beijing Ditan Hospital from October 31, 2019 to October 31, 2021 and could not undergo surgery or experienced metastasis after surgery. All patients were treated with lenvatinib combined with sintilimab as the second-line therapy. The patients were followed up, and the RECIST1.1 criteria were used to assess treatment outcome. The primary endpoint was time to progression (TTP), and the secondary endpoints were tumor objective response rate (ORR), disease control rate (DCR), overall survival (OS) time, and safety. The Kaplan-Meier method was used to plot survival curves, and the log-rank test was used for comparison between groups. Results A total of 27 patients were enrolled, among whom there were15 male patients (55.6%) and 12 female patients (44.4%), with a median age of 58 years (range 33-73 years). The median TTP for these patients was 5.5 (95% confidence interval [ CI ]: 1.7-9.3) months, and 13 patients (48.1%) died of disease progression, with a median OS time of 11.2 (95% CI : 5.0-17.4) months. The overall ORR and DCR were 40.7% and 70.3%, respectively. Of all patients, 66.7% experienced varying degrees of adverse events, and among these patients, 44.4% had an increase in alanine aminotransferase, 44.4% had an increase in aspartate aminotransferase, 37.0% had hypertension, 29.6% had an increase in bilirubin, 29.6% experienced diarrhea, and 25.9% each experienced proteinuria, anorexia, and weakness. No treatment-related death was observed, and only 1 patient developed grade Ⅳ immune-related hepatotoxicity and was relieved without sequelae after corticosteroid therapy, resulting in permanent withdrawal of sintilimab. The patients with lymph node metastasis had a significantly shorter median TTP than those without lymph node metastasis (4.5 months vs 18.8 months, P =0.035), and the patients who achieved disease remission had a significantly longer median TTP [11.6 months (95% CI : 5.6-17.6) vs 2.8 months (95% CI : 1.8-3.8), P < 0.001]; the patients with lymph node metastasis had a shorter median OS time [9.6 months (95% CI: 7.9-11.3) vs 21.9 months (95% CI : 0-44.9), P =0.053], and the patients who achieved disease remission had a significantly longer median OS time [16.6 months (95% CI : 9.0-24.2) vs 6.9 months (95% CI : 3.6-10.2), P =0.011]. Conclusion Lenvatinib combined with sintilimab has a marked clinical effect and a low incidence rate of serious adverse events as the second-line therapy for advanced ICC, and therefore, it is a safe and effective treatment regimen.

Clinical effect of programmed cell death-1 inhibitor combined with lenvatinib in treatment of advanced primary liver cancer and related adverse events / 临床肝胆病杂志

ObjectiveTo investigate the clinical effect of domestic programmed cell death-1 (PD-1) inhibitor combined with lenvatinib in the treatment of advanced primary liver cancer and related adverse events. MethodsA retrospective analysis was performed for the clinical data of 24 patients with advanced primary liver cancer who were treated with domestic PD-1 inhibitor combined with lenvatinib in Beijing Ditan Hospital, Capital Medical University, from January 1, 2019 to April 2, 2020, with 15 patients in the Camrelizumab+lenvatinib group, 7 patients in the Sintilimab+lenvatinib group, and 2 patients in the Toripalimab+lenvatinib group. During follow-up, Modified Response Evaluation Criteria in Solid Tumors was used to evaluate the treatment outcome of intrahepatic lesions, and RECIST1.1 was used to evaluate extrahepatic metastatic lesions. The Kaplan-Meier method was used to evaluate survival time. ResultsAmong the 24 treatment-experienced patients, 11 achieved partial response, 7 achieved a stable disease, and 6 had disease progression, resulting in an objective response rate of 45.8% and a disease control rate of 75.0%. The median time to disease progression was 8.4 (95% confidence interval: 6.89-9.91) months. The incidence rate of adverse events was 54.17%, and the most common adverse events were fatigue (29.17%) and hypertension (25.00%). ConclusionPD-1 inhibitor combined with lenvatinib has a marked clinical effect in the treatment of advanced primary liver cancer, with a low incidence rate of serious adverse events, and thus it is a safe and effective treatment regimen.

Related Factors Affecting Long-term Prognosis of AML Children with Positive RUNX1-RUNX1T1 / 中国实验血液学杂志

OBJECTIVE@#To analyze the related factors affecting the long-term prognosis of acute myeloid leukemia (AML) children with positive RUNX1-RUNX1T1.@*METHODS@#The clinical data of 63 chlidren with positive RUNX1-RUNX1T1 AML treated by BCH-AML 05 regimen in our hospital from January 2010 to December 2015 were collected and analyzed retrospectively. The level of RUNX1-RUNX1T1 was detected at the time of initial diagnosis (T), after the first induction treatment (T), after the second induction treatment (T), after the first consolidation treatment (T), after the second consolidation treatment (T) and after the third consolidation treatment (T). According to the fusion transcript levels of RUNX1-RUNX1T1 the AML children were divided into low-expression group and high-expression group; the threshold values for grouping were 10 copies/10 β-glucuronidase (GUS), 10 copies/10 GUS, 10 copies/10 GUS, 10 copies/10 GUS, 1 copies/10 GUS and 0 copies respectively. The gained data were enrolled in the statistical analysis.@*RESULTS@#23 cases of 63 children died during the follow-up period, and the median follow-up time of the remaining 40 children were 30.04 (11-60) months. There were statistically significant differences in CD15 positive rate between low-expression group and high-expression group (P0.05). Univariate analysis showed that sex, Plt counts at T and fusion transcript levels at T, T and T correlated with the 5-year overall survival rate (P10 copies/10 GUS at T was an independent risk factor for 5-year overall survival rate (HR=2.13, 95%CI: 1.04-7.78)(P<0.05).@*CONCLUSION@#The fusion transcript level after the first induction therapy in RUNX1-RUNX1T1-positive AML children is an independent factor influencing the long-term prognosis.

Salvianolic acids improve liver lipid metabolism in ovariectomized rats via blocking STAT-3/SREBP1 signaling / 中国天然药物

Postmenopausal women, who have reduced circulating estrogen levels, are more prone to develop obesity and related metabolic diseases than premenopausal women. The absence of safe and effective treatments for postmenopausal obesity has changed the focus to natural products as alternative remedies. Total salvianolic acids (TSA) are the major water-soluble ingredients of Danshen. Salvianolic acid (SA) is the major constituent of the TSA. Salvianolic acids, including TSA and SA, are widely used in traditional Chinese medicine. In the present study, ovariectomized rats and LO2 cells were used to study the effects of salvianolic acids on body weight gain and hepatic steatosis. Salvianolic acids reduced ovariectomy (OVX)-induced body weight gain, attenuated the expressions of hepatic lipogenic genes, such as sterol regulatory element binding protein (SREBP)1, fatty acid synthase (FAS), and stearoyl-CoA desaturase (SCD)1, and decreased the liver triglyceride (TG) and total cholesterol (TC). For the molecular mechanisms, OVX and high glucose-induced phosphorylation of signal transducer and activator of transcription (STAT)-3 was inhibited by salvianolic acids treatment. In LO2 cells, inhibition of STAT-3 by siRNA attenuated the increased expression of SREBP1 and TG induced by high glucose. Salvianolic acids reduced the upregulation of SREBP1 and TG induced by high glucose in LO2 cells. In conclusion, these findings illustrated that salvianolic acids markedly alleviated the lipid metabolism disorders and protected against the postmenopausal obesity. The underlying mechanism was probably associated with the regulation of STAT-3 signaling.

Salvianolic acids improve liver lipid metabolism in ovariectomized rats via blocking STAT-3/SREBP1 signaling / 中国天然药物

Postmenopausal women, who have reduced circulating estrogen levels, are more prone to develop obesity and related metabolic diseases than premenopausal women. The absence of safe and effective treatments for postmenopausal obesity has changed the focus to natural products as alternative remedies. Total salvianolic acids (TSA) are the major water-soluble ingredients of Danshen. Salvianolic acid (SA) is the major constituent of the TSA. Salvianolic acids, including TSA and SA, are widely used in traditional Chinese medicine. In the present study, ovariectomized rats and LO2 cells were used to study the effects of salvianolic acids on body weight gain and hepatic steatosis. Salvianolic acids reduced ovariectomy (OVX)-induced body weight gain, attenuated the expressions of hepatic lipogenic genes, such as sterol regulatory element binding protein (SREBP)1, fatty acid synthase (FAS), and stearoyl-CoA desaturase (SCD)1, and decreased the liver triglyceride (TG) and total cholesterol (TC). For the molecular mechanisms, OVX and high glucose-induced phosphorylation of signal transducer and activator of transcription (STAT)-3 was inhibited by salvianolic acids treatment. In LO2 cells, inhibition of STAT-3 by siRNA attenuated the increased expression of SREBP1 and TG induced by high glucose. Salvianolic acids reduced the upregulation of SREBP1 and TG induced by high glucose in LO2 cells. In conclusion, these findings illustrated that salvianolic acids markedly alleviated the lipid metabolism disorders and protected against the postmenopausal obesity. The underlying mechanism was probably associated with the regulation of STAT-3 signaling.

Basic medical insurance for the aged from Zhoupu communities in Shanghai: Analysis of satisfaction and influencing factors / 第二军医大学学报

Objective To investigate satisfactory degree on basic medical insurance system for the aged and to provide more data for improving insurance policies. Methods A face-to-face questionnaire survey was carried out with 367 elderly residents in Zhoupu communities in Shanghai using cluster random sampling method. The survey included social demographic characteristics, medical insurance, chronic disease, health cost, etc. The data were analyzed using descriptive statistical analysis, Chi-square test and multivariate logistic regression analysis by SPSS software. Results Totally 357 effective questionnaires were collected, with the effective rate being 97.3%. The overall satisfaction rate of the retired employees was high according to our survey, but the satisfaction rates of urban non-employed residents and land-requisitioned peasants were relatively low (P<0.001). Single factor analysis and multiple factor analysis both revealed that income, number of chronic diseases and self-rating health had significant association with satisfaction rate of the participants (P<0.05). Conclusion The basic medical insurance for urban non-employed residents and landless peasants needs to be further improved.

Application of a refined fibrosis staging system to evaluate therapeutic efficacy in chronic viral hepatitis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To generate a refined staging system of fibrosis in chronic viral hepatitis and to assess its accuracy and sensitivity for evaluating therapeutic efficacy of anti-fibrosis drug treatments.</p><p><b>METHODS</b>A refined fibrosis staging system was established according to the detailed characteristics of progressive fibrosis. A total of 396 liver puncture biopsy specimens were collected from patients before and after anti-fibrosis therapy and used to evaluate the refined staging system. According to the original fibrosis staging system and refined fibrosis staging system, fibrosis staging differences from before and after treatment were analyzed by Chi-squared test and paired-samples t-test respectively.</p><p><b>RESULTS</b>The refined fibrosis staging system detected significant changes in fibrosis stage that occurred in response to treatment (before treatment: 6.55 +/- 2.93 vs. after treatment: 6.19 +/- 2.945, P less than 0.01). However, the original (unrefined) staging system was unable to differentiate therapy-related changes in fibrosis stage (x2= 3.144, P = 0.534).</p><p><b>CONCLUSION</b>The newly-developed refined fibrosis staging system was able to effectively evaluate the therapeutic efficacy of anti-fibrosis drug treatment and performed better than the original staging system.</p>

Dynamic contrast-enhanced MSCT findings of intraductal papillary neoplasm of the bile ducts / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the findings of contrast-enhanced multislice computed tomography (MSCT) that characterize intraductal papillary neoplasms of bile ducts (IPNB).</p><p><b>METHODS</b>The MSCT findings and clinical data of 16 cases of IPNB proven by surgical pathology were reviewed retrospectively.</p><p><b>RESULTS</b>Among the 16 cases, nine were adenoma (multi-lesions, n = 5; single lesions, n = 4) and seven were adenocarcinoma (multi-lesions, n = 4; single lesions, n = 3). Among the nine adenoma cases, seven showed nodules or masses in the expanding intrahepatic bile ducts with asymmetrical low density on plain scan, and two showed obvious expansion of biliary ducts and the inner wall of bile ducts was rough. All seven of the adenocarcinoma cases showed nodules or masses in the expanding intrahepatic bile ducts with asymmetrical low density-like adenoma. When contrast enhancement was applied, the nine adenoma cases manifested slight-to-moderate degrees of asymmetrical enhancement. For the seven adenocarcinoma cases, two showed asymmetrical enhancement similar to that of the adenoma cases and five showed continued enhancement; one case showed malignant infiltration of the bile duct and evident damage in the adjacent hepatic tissue. The CT plain scan findings for the two groups (adenoma and adenocarcinoma) were not significantly different (t = -1.17, P = 0.2632). Significantly different findings were obtained with the MSCT imaging analysis for the arterial phase (t = 6.53, P less than 0.01) and the portal vein phase (t = 5.63, P less than 0.01). All cases showed asymmetrical expansion of intrahepatic biliary ducts, diffuse or local, and four cases showed moderate expansion of the common bile duct. One adenocarcinoma case showed intumescence in the celiac lymph node by moderate asymmetrical enhancement.</p><p><b>CONCLUSION</b>MSCT is helpful for the differential diagnosis of IPNB from other hepatic lesions.</p>

Effects of different artificial dermal scaffolds on vascularization and scar formation of wounds in pigs with full-thickness burn / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effects of three kinds of artificial dermal scaffolds on vascularization and scar formation of wounds in pigs with full-thickness burn.</p><p><b>METHODS</b>Eighteen Bama miniature pigs were divided into chitosan scaffold (CS) group, sulfonated carboxymethyl chitosan scaffold (SCCS) group, and acellular dermal matrix (ADM) scaffold group according to the random number table, with 6 pigs in each group. Every pig in all groups was inflicted with 4 or 8 full-thickness scald wounds on the back (totally 96 wounds). Forty-eight hours after injury, eschars of all wounds were excised. Twenty-four wounds in CS group were transplanted with double-layer artificial dermis of collagen-chitosan and silicone rubber, those in SCCS group with double-layer artificial dermis of collagen-sulfonated carboxymethyl chitosan and silicone rubber, and those in ADM scaffold group with ADM. The rest 24 wounds in the three groups were dressed with vaseline gauze as control group. After 2 weeks of treatment, all wounds of every group were covered with skin. In post treatment (scaffold transplantation or gauze covering) week (PTW) 1, 2, 3, and 4, gross condition of wound was observed, and specimens from central parts of wounds were harvested for observation and assessment of vessels or cells with positive expression of CD31, α smooth muscle actin (α-SMA), TGF-β(1) and TGF-β(3) with SP staining. Data were processed with one-way analysis of variance and LSD test.</p><p><b>RESULTS</b>(1) Degree of vascularization in SCCS group was better than that in the other three groups. (2) The number of vessels with positive expression of CD31 in CS, SCCS, ADM scaffold, and control groups increased gradually from PTW 1 to PTW 3, and decreased in PTW 4. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 24.005, 38.822, 25.274, 3.856, P < 0.05 or P < 0.01). The numbers of vessels that expressed CD31 in SCCS group from PTW 1 to PTW 3 were more than those in the other three groups (with P values all below 0.05). (3) The numbers of vessels that expressed α-SMA in CS, SCCS, and ADM scaffold groups from PTW 1 to PTW 3 showed the similar trend of change to those of vessels that expressed CD31, which increased gradually in control group from PTW 1 to PTW 4. There were obvious differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 22.637, 28.087, 62.651, 18.055, P values all below 0.01). The number of vessels that expressed α-SMA in SCCS group from PTW 1 to PTW 4 was more than that in the other three groups (with P values all below 0.05). (4) From PTW 1 to PTW 4, the number of cells with expression of TGF-β(1) in CS group was respectively (127 ± 8), (167 ± 19), (170 ± 18), (144 ± 10) per 400 times visual field, that in SCCS group was respectively (171 ± 17), (207 ± 25), (130 ± 30), (69 ± 16) per 400 times visual field, that in ADM scaffold group was respectively (106 ± 8), (159 ± 17), (171 ± 11), (145 ± 11) per 400 times visual field, and that in control group was respectively (100 ± 20), (150 ± 18), (200 ± 14), (172 ± 20) per 400 times visual field. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 29.675, 9.503, 13.107, 54.515, P values all below 0.01). Compared with those in SCCS group, the number of cells that expressed TGF-β(1) in the other three groups was decreased in PTW 1, 2 but increased in PTW 3, 4 (with P values all below 0.05). (5) The number of cells that expressed TGF-β(3) in 4 groups increased gradually from PTW 1 to PTW 3, and decreased or increased continually in PTW 4. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 140.612, 945.850, 714.037, 119.147, P values all below 0.01). The number of cells with positive expression of TGF-β(3) in SCCS group from PTW 1 to PTW 4 was more than that in the other three groups (with P values all below 0.05).</p><p><b>CONCLUSIONS</b>The collagen-sulfonated carboxymethyl chitosan dermal scaffold can rapidly induce growth and maturation of blood vessels during wound healing after burn. It is beneficial for wound repair at early stage with inhibition of scar proliferation.</p>

A two-year follow-up study of 58 patients with postpartum thyroiditis / 中华内分泌代谢杂志

ObjectiveTo investigate the cumulative incidence of persistent hypothyroidism in patients who were diagnosed as postpartum thyroiditis ( PPT),and to determine the factors associated with the development of persistent hypothyroidism in those patients.MethodsThe present study was performed as the continuous study followed by the former epidemiological survey on PPT,in which 58 patients with PPT (35 overt PPT and 23 subclinical PPT) were diagnosed.The 58 patients were followed up at 12th month postpartum,and then for every 6 months until 24 months postpartum.Fasting blood samples were taken for testing serum TSH,thyroid peroxidase antibody ( TPOAb),and thyroglobulin antibody ( TgAb ).Free T3 ( FT3 ),free T4 ( FT4 ),and TSH receptor antibody ( TRAh ) were detected if TSH was abnormal.50 healthy postpartum women were used as control group.ResultsOf the total 58 PPT patients,91,4％ ( n =53 ) were successfully followed.Five patients with overt PPT and 6 patients with subclinical PPT developed persistent hypothyroidism,and the cumulative incidence of persistent hypothyroidism in the studied PPT patients was 20.8％.Among 15 PPT patients who had a classical biphasic course (a thyrotoxic phase followed by a hypothyroid phase),persistent hypothyroidism was seen in 26.7％ (n =4 ).Among 11 PPT patients with hypothyroidism only,persistent hypothyroidism was seen in 63.6％ ( n =7).On the contrary,none of the patients with thyrotoxicosis only had persistent hypothyroidism.All of the patients who developed persistent hypothyrodism had a higher TSH levels than 4.8 mU/L at 6th month postpartum.Before delivery,TSH levels of the patients developed persistent hypothyroidism were significantly higher than those of the patients with transient hypothyroidism,and this was the case at the 12th month postpartum ( all P＜0.01 ).PPT patients maintained a relatively higher rate of thyroid autoantibodies.The positive rate of TPOAb at the 12th,18th,and 24th month postpartum was 56.6％,50.9％,and 52.8％,respectively; and the positive rate of TgAb being 35.8％,30.2％,and 30.2％,respectively.Both the positive rate and titer of TPOAb in patients with overt PPT were higher than those in patients with subclinical PPT at the 18th and 24th month postpartum (P＜0.05).Conclusions 20.8％ patients with PPT developed persistent hypothyroidism at the 24th month postpartum.Whether a patient with PPT would develop persistent hypothyroidism depends on his clinical feature and TSH level.

Analysis of mental health status and its influencing factors of the elderly living in xi'an, China / 第二军医大学学报

Objective To analyze the mental health status and its influencing factors in the elderly living in Xi'an communities, so as to bring forth countermeasures to improve their mental health. Methods Convenience sampling was used to select 310 residents aged 60 years or above from 2 communities in Xi'an, and the subjects were surveyed by a self-designed questionnaire. The survey included social demographic characteristics, social support, disability, chronicdisease costs, physical exercises, mental health problems and the coping methods. Univariate chi-square test and multivariable logistic regression were used to identify factors associated with mental health of the elderly. Results Questionnaires with uncompleted answersor suspected unreal answers were excluded. Totally 304 effective questionnaires were collected, with the effective rate being 98. 1%. The prevalence of mental disorderswas 27. 1 % in the present study. Chi-square tests showed that the chronicdisease costs, disability, social support and physical exercises were the influencing factors of the mental health status. Multivariable logistic regression analysis demonstrated that medical insurance, chronicdisease costs, social support and physical exercises were important factors influencing themental health of the elderly. Conclusion It is important to emphasize the spiritual support for the aging population and the fading family support in China. We should improve the social support for the elderly and build up mental supporting services so as to improve their mental health.

Analysis of mental health status and its influencing factors of the elderly living in xi'an, China / 第二军医大学学报

Objective To analyze the mental health status and its influencing factors in the elderly living in Xi'an communities, so as to bring forth countermeasures to improve their mental health. Methods Convenience sampling was used to select 310 residents aged 60 years or above from 2 communities in Xi'an, and the subjects were surveyed by a self-designed questionnaire. The survey included social demographic characteristics, social support, disability, chronicdisease costs, physical exercises, mental health problems and the coping methods. Univariate chi-square test and multivariable logistic regression were used to identify factors associated with mental health of the elderly. Results Questionnaires with uncompleted answersor suspected unreal answers were excluded. Totally 304 effective questionnaires were collected, with the effective rate being 98. 1%. The prevalence of mental disorderswas 27. 1 % in the present study. Chi-square tests showed that the chronicdisease costs, disability, social support and physical exercises were the influencing factors of the mental health status. Multivariable logistic regression analysis demonstrated that medical insurance, chronicdisease costs, social support and physical exercises were important factors influencing themental health of the elderly. Conclusion It is important to emphasize the spiritual support for the aging population and the fading family support in China. We should improve the social support for the elderly and build up mental supporting services so as to improve their mental health.

Pooled sampling method under given sensitivity and specificity in improving accuracy of point estimator of population rate / 第二军医大学学报

Objective: To use the pooled sampling method to reduce the error caused by the detecting precision, so as to improve the accuracy of population rate estimates. Methods: A formula was deduced on how to obtain the ideal rate under given sensitivity and specificity, and the mean relative errors of rate estimation with different pool sizes were simulated and calculated using software SAS 9.1. Results: When the actual rate was lower than the ideal rate, the errors caused by the detecting precision could be greatly reduced through adjusting the rate of mixed samples. Conclusion: When the population rate is low, the accuracy of rate estimation can be improved and the numbers of tests can be reduced by using pooled sampling method under given specificity and sensitivity.

PD-1/PD-L1 expressions in liver tissues of patients with chronic HBV infection / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To detect and compare the PD-1/PD-L1 (programmed death 1/programmed death 1 ligand) expressions in the liver tissues of chronic HBV infection patients in immune tolerant phase and those in immune clearance phase.</p><p><b>METHODS</b>Liver biopsy samples were divided into two groups: 25 samples from patients in immune clearance phase and 19 samples from patients in immune tolerant phase. PD-1/PD-L1 expressions on T lymphocytes in these liver biopsy specimens were detected by immunohistochemistry method. Percentage of PD-1/PD-L1 positive cells among CD3 positive cells was calculated by semi-quantitative evaluation. Differences between the two groups were statistically analyzed.</p><p><b>RESULTS</b>PD-1/PD-L1 expressions were significantly higher in the patients in immune tolerant phase as compared to that in immune active phase (P < 0.05). No statistical difference found between the two groups for PD-L1 expression in Kupffer cells (P > 0.05).</p><p><b>CONCLUSION</b>PD-1/PD-L1 expression level can reflect the immune functions of chronic hepatitis B patients.</p>

Comparative study on repair of full-thickness burn wound with different artificial dermal stent in pigs / 中华整形外科杂志

<p><b>OBJECTIVE</b>To compare differences of angiogenesis among collagen- chitosan, collagen-sulfonated carboxymethyl chitosan porous scaffolds and acellular dermal matrix after these three different scaffolds with silicone membrane were transplanted on the wounds of full thickness burn, and the wound repair of different scaffolds with epidermis grafting on.</p><p><b>METHODS</b>Angiogenesis in different dermal scaffolds, the wound surface and epidermis survival were observed in 1, 2, and 3 weeks after the three different scaffolds were respectively transplanted on wounds of full thickness burn with debridement in 6 Bama miniature pigs (total 18 pigs in 3 groups). At the same time, CD34 positive signals (neo-forming microvessels) were detected by immunohistochemical staining. The wounds without any scaffold transplantation were studied as the control.</p><p><b>RESULTS</b>Angiogenesis had been fundamentally finished in 2 weeks after implantation of collagen- sulfonated carboxymethyl chitosan porous scaffold. And fundamental angiogenesis in collagen- chitosan porous scaffolds and acellular dermal matrix needed at least 3 weeks. Neo-forming micro-vessels perpendicular to wound beds with these three different scaffolds were more than those in the control wounds without scaffold. CD34 positive signals (neo-forming micro-vessels) were significantly higher in wounds at the second week than those in wounds at the first week. And those in wounds at the third week were significantly higher than those in wounds at the second week in all wounds with different scaffold transplantations and the control wounds. CD34 positive signals in the group of sulfonated carboxymethyl chitosan porous scaffold on the 1st, 2nd and 3rd week after the scaffold transplantation were significantly higher than those corresponding signals in the other three groups. Epidermis on the sulfonated carboxymethyl chitosan porous scaffold which had been transplanted on burn wound for 1 week could survive perfectly, however, epidermis on the collagen- chitosan porous scaffold or acellular dermal matrix could not survive until these two scaffolds had been transplanted on the burn wounds for at least 2 weeks.</p><p><b>CONCLUSIONS</b>These three different scaffolds could repair the full thickness skin defects caused by burn, and angiogenesis of sulfonated carboxymethyl chitosan porous scaffold is the best.</p>

Differences of wound contraction and apoptosis in full-thickness burn wounds repaired with different artificial dermal stent in pigs / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the roles and differences of angiogenesis of different dermal scaffolds on wound contraction and apoptosis during full-thickness burn wound repair.</p><p><b>METHODS</b>Wounds were observed at different time after the collagen-sulfonated carboxymethyl chitosan porous scaffold or collagen-chitosan porous scaffold or acellular dermal matrix were respectively transplanted on wounds of full thickness burn with debridement in Bama miniature pigs. At the same time, vessels and myo-fibroblasts expressing α-smooth muscle action(α-SMA) and apoptosis in wounds of different time were detected in situ by immunohistochemical staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling. The burn wounds without any scaffold transplantation were studied as the control.</p><p><b>RESULTS</b>Wounds with different scaffolds transplantation were different from granulation wounds. Vessels expressing α-SMA had been increasing continuously in the wounds from 1 to 3 weeks after different scaffolds transplantation and decreased in wounds after epidermis had been grafted for 2 weeks on surface of the scaffolds transplanted on wounds for 2 weeks. Vessels expressing α-SMA were the most in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the least in the control wounds without dermal scaffold at different time. Myo-fibroblasts expressing α-SMA was the least in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the peak of expressions was on the 2nd week, however, the peak in the wounds with the other two scaffolds transplantation and in the control wound without dermal scaffold was on the 3rd week. Myo-fibroblasts expressing α-SMA was the most in the control wounds. Apoptosis had been increasing continuously in the transplantation wounds from 2 to 4 weeks after different scaffolds transplantation, however, apoptosis had begun to increase continuously from 3 to 4 weeks in the control wounds. Apoptosis was the most in the wounds with collagen-sulfonated carboxymethyl chitosan porous scaffold transplantation and the least in the control wounds without dermal scaffold from 3 to 4 weeks.</p><p><b>CONCLUSION</b>Sulfonated carboxymethyl chitosan can promote migration of reparative cells and angiogenesis, and it can repair full-thickness burn wound fast and well.</p>

Optimized design for in vitro experiment of electret transdermal patches / 第二军医大学学报

Objective: Currently most researches concerning electret transdermal patches are performed by one-factor experimental design with relatively poor efficiency. To analyze the influencing factors in a more systematical and effective manner, we put forward an optimized design for in vitro experiment with electret transdermal patches. Methods: Due to the great number and various levels of factors impacting the charge storage stability of electrets, we combined uniform design and orthogonal design to optimize the screening for the stable transdermal patch. In the following in vitro study, we optimized the design of percutaneous experiments to screen for charge polarity and penetration enhancers using a two-factor experimental design. And using an orthogonal experimental design, we further studied the influence of different levels of main drug, surface potential and penetration enhancer on transdermal absorption. Conclusion: The present design, taking into consideration of various factors and using multi-factor experimental design, can more effectively analyze data, reduce workload, and is feasible.

Comparison of the mechanisms of intralesional steroid, interferon or verapamil injection in the treatment of proliferative scars / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of intralesional steroid, interferon alpha-2b or verapamil injection on proliferation, apoptosis and TGF-beta1 expression in keloid and hypertrophic scar in vivo.</p><p><b>METHODS</b>6 patients with keloids and 6 patients with hypertrophic scar were treated with intralesional injection of triamcinolone acetonide (40 mg/ml) or IFN alpha-2b (15 x 10(5) U/ml) or verapamil (2.5 mg/ml). Samples were collected on the 7th day after intralesional injection. Samples of untreated keloid and hypertrophic scar and normal skin were used as control. Expression of PCNA and TGF-beta1 was detected in situ by immunohistochemical staining, and apoptosis was detected in situ by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL).</p><p><b>RESULTS</b>1) Triamcinolone acetonide could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, as well as inducing apoptosis. 2) IFN alpha-2b could prohibit proliferative scars through inhibiting cell proliferation and TGF-beta1 expression, but not inducing apoptosis; 3) Verapamil could also prohibit proliferative scars through inhibiting proliferation and TGF-beta1 expression in fibroblasts, as well as inducing apoptosis. While the effect of inducing apoptosis was stronger than that of triamcinolone acetonide, the effect of inhibiting TGF-beta1 expression was weaker than those of triamcinolone acetonide and IFN alpha-2b.</p><p><b>CONCLUSIONS</b>Although intraleional injection of steroid, interferon alpha-2b or verapamil were all effective in the treatment of keloid and hypertrophic scar, their mechanisms are not similar.</p>

Study on repair of full-thickness skin defect with collagen-chitosan dermal stent in pigs / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate angiogenesis of collagen-chitosan porous scaffold, and to study survive of skin grafts on the scaffold after bilayer dermal equivalent (BDE) was transplanted on wounds with full thickness skin defects.</p><p><b>METHODS</b>The full thickness skin defects were made on 10 Bama miniature pigs and the BDE composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wound. Angiogenesis in dermal equivalent, wound healing, and healing and survive of skin grafts on dermal equivalent were observed in 1, 2, and 3 weeks after the BDE transplantation. At the same time, CD34 positive signals (neo-forming micro-vessels) were detected by immunohistochemical staining.</p><p><b>RESULTS</b>Inflammatory cells and fibroblasts infiltrated into dermal equivalent and a few new micro-vessels had been formed in 1 week after the BDE transplantation; neo-forming micro-vessels perpendicular to wound bed had increased significantly in 2 weeks after the BDE transplantation; neo-forming micro-vessels could be observed in almost all dermal equivalents in 3 weeks after the BDE transplantation. CD34 positive signals (neo-forming micro-vessels) in 3 weeks after the BDE transplantation was much more than those in 2 weeks after the BDE transplantation, and CD34 positive signals in 2 weeks after the BDE transplantation was much more than those in 1 week after the BDE transplantation. Survival rate of intermediate split thickness skin graft were 10%, 70% and 100% respectively after the skin grafts had been grafted for 2 weeks on surface of the scaffold which had been transplanted for 1, 2 and 3 weeks. Epidermis which had been grafted on surface of the scaffold for 1 or 2 weeks could perfectly survive after BDE had been transplanted for 1 or 2 weeks.</p><p><b>CONCLUSIONS</b>Collagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect and can induce fibroblast infiltration and new micro-vessel formation. Epidermis grafted on surface of collagen-chitosan porous scaffold can perfectly repair wounds, and it has brilliant applied prospects in repairing skin defect.</p>