AIFM1 variants associated with auditory neuropathy spectrum disorder cause apoptosis due to impaired apoptosis-inducing factor dimerization / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Systematic evaluation of the incidence of the knee donor area after autobone cartilage mosaic xentoplasty / 中国骨伤

OBJECTIVE@#To provide an overview of the incidence of knee donor -site morbidity after autologous osteochondral mosaicplasty.@*METHODS@#A comprehensive search was conducted in PubMed, EMbase, Wanfang Medical Network, and CNKI databases from January 2010 to April 20, 2021. Relevant literature was selected based on predefined inclusion and exclusion criteria, and data were evaluated and extracted. The correlation between the number and size of transplanted osteochondral columns and donor-site morbidity was analyzed.@*RESULTS@#A total of 13 literatures were included, comprising a total of 661 patients. Statistical analysis revealed an incidence of knee donor-site morbidity at 8.6% (57/661), with knee pain being the most common complaint, accounting for 4.2%(28/661). There was no significant correlation between the number of osteochondral columns and postoperative donor-site incidence (P=0.424, N=10), nor between the diameter size of osteochondral columns and postoperative donor-site incidence(P=0.699, N=7).@*CONCLUSION@#Autologous osteochondral mosaicplasty is associated with a considerable incidence of knee donor-site morbidity, with knee pain being the most frequent complaint. There is no apparent correlation between donor-site incidence and the number and size of transplanted osteochondral columns. Donors should be informed about the potential risks.

Time-course changes of intraosseous environment and shear resistance mechanical properties of jawbone in SD rats with fluorosis / 环境与职业医学

Background At present, a large number of reports focus on the bones of limbs and trunk, while there are few studies on the effect of fluorosis on jawbone which is the inevitable structural basis for the development and treatment of oral diseases. Objective To preliminarily investigate the effect of fluoride exposure on the mechanical properties of jawbone by observing the changes in the intraosseous environment and the maximum load against shearing force (LSFmax) of the jawbone in rats with chronic fluoride treatment. Methods Screening experiment: 48 SD male rats were randomly divided into a control group and three fluoride exposure groups (50, 150, and 250 mg·L−1 fluoride concentration), 12 rats in each group. The fluoride exposure groups were molded by feeding different concentrations of sodium fluoride solution, and the control group drank tap water from Guizhou area. Each group was further divided into 4 subgroups with 3 animals each according to observation time points after 0, 2, 4, and 6 months. The LSFmax of the jawbone was measured with an electronic universal ergometer, the expression of type I collagen (Col1) was shown by Sirius red staining, and the expression of runt-related transcription factor 2 (Runx2) was determined semi-quantitatively by immunohistochemistry at selected time points. Formal experiment: 12 male SD rats were randomly divided into a fluoride exposure group and a control group. The fluoride exposure group were fed with 150 mg·L−1 sodium fluoride solution, and the control group drank tap water from Guizhou. After feeding with fluoride for 5 months, the ergometer was used to measure the LSFmax of the jawbone. Osteoclasts were counted after tartrate resistant acid phosphatase (TRAP) staining. Col1, Runx2, bone morphogenetic protein 2 (BMP-2), alkaline phosphatase (ALP), and cathepsin K (Cath K) were detected semi-quantitatively by immunohistochemistry expression and Sirius red staining. Micro computed tomography (Micro CT) was used to observe the trabecular bone microstructure. Results Screening experiment: The LSFmax of the control group and the 50 mg·L−1 fluoride exposure group reached the peak value at the 2nd month, and the LSFmax of the 50 mg·L−1 fluoride exposure group reached the valley value at the 4th month. The LSFmax of the 150 mg·L−1 fluoride exposure group at the 4th month was higher than that at the 6th month (P<0.05). There was no significant difference in the LSFmax at each time point in the 250 mg·L−1 fluoride exposure group. At the same time point, there was no statistically significant difference in LSFmax among the groups. The Col1 levels of the 50 mg·L−1, 150 mg·L−1, and 250 mg·L−1 fluoride exposure groups were higher than the time point 0 from the 2nd month (P＜0.05). The Runx2 showed no statistically significant difference by concentration or time. Formal experiment: After feeding with 150 mg·L−1 fluoride for 5 months, the LSFmax of the fluoride exposure group was greater than that of the control group (P<0.05). The expressions of Col1, Runx2, BMP2, ALP, and Cath K in the fluorosis exposure group were higher than those in the control group (P<0.05). There were no statistically significant differences in osteoclast count or indicators of bone trabecular microstructure. Conclusion Chronic fluoride exposure may increase the shear strength of jaw bone.

Analysis of the glycosylation heterogeneity of recombinant human pro-urokinase using UPLC-MS / 药学学报

The glycosylation heterogeneity of recombinant human pro-urokinase (pro-UK) was assessed using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Firstly, the source of heterogeneity was determined by measuring the Mr of intact protein before and after N-deglycosylation. Glycosylation sites and the proportion of O-glycopeptides then were determined at the peptide level. Finally, the N-glycans were confirmed and quantified using the N-glycan profile. Results show that the structural heterogeneity of pro-UK is mainly caused by glycosylation. All T18 were fucosylated, and 6.4% of S138/139 was O-glycosylated with two kinds of oligosaccharides with a ratio of 6.0% and 0.4% respectively. All N302 positions were N-glycosylated by more than ten types of glycans, among which A2F and A3F accounted for 80% of the total. The assessment of glycosylation heterogeneity of pro-UK will provide a reference for quality standardization.

Investigation of Decoction Process of Digda-4 Decoction / 中国实验方剂学杂志

Objective:To optimize the decoction process of Digda-4 decoction(DGD-4D), and provide reference for the standardization study of decoction of Mongolian medicine decoction. Method:Taking DGD-4D as model drug, different decoction methods of Mongolian medicine were compared, HPLC was used to determine contents of aesculetin, geniposide, picroside Ⅰ and picroside Ⅱ.On the basis of single factor tests, central composite design-response surface methodology was adopted to optimize the decoction process of DGD-4D with transfer rates of 4 components and dry extract rate as indexes, regression model fitting was carried out by Design-Expert 8.0.6 software, prediction model of process parameters was established, and the optimal process was verified. Result:The optimal decoction condition of DGD-4D was determined to be adding 40 times the amount of water and decocting for 17 min, decocting once.Transfer rates of aesculetin, geniposide, picroside Ⅰ, picroside Ⅱ and dry extract rate were 70.01%, 94.11%, 61.23%, 92.32%, 32.89%, respectively. Conclusion:The optimum decoction process of DGD-4D is established, it has important reference significance for excavating, sorting, improving the level of Mongolian medicine preparations and ensuring the consistency of their clinical efficacy.

Development of an Internally Controlled Reverse Transcription Recombinase-aided Amplification Assay for the Rapid and Visual Detection of West Nile Virus / 生物医学与环境科学（英文）

West Nile virus (WNV) causes West Nile fever and West Nile encephalitis. Because infection by WNV creates serious public health problems, its simple, rapid, and visual detection is very important in clinical practice, especially in resource-limited laboratories. We have developed a rapid, specific, and highly sensitive internally controlled reverse transcription recombinase-aided amplification (RTRAA) assay to detect WNV, using both real-time fluorescence and the lateral flow dipstick (LFD) at 39.0 °C for 30 min. The analytical sensitivity of the RT-RAA assay was 10 plasmid copies and 1.6 pfu per reaction with real-time fluorescence, and 1,000 plasmid copies per reaction with the LFD. No crossreaction with other control viruses was observed. Compared with the RT-qPCR assay, the RT-RAA assay demonstrated 100% sensitivity and 100% specificity for WNV.

Research on extraction process of Digeda-4 flavored decoction based on QbD concept / 中国中药杂志

To establish and validate the design space of the Digeda-4 flavored decoction( DGD-4D) extraction process by using the quality by design( Qb D) concept. With DGD-4D decoction pieces as a model drug,with the transfer rate of aesculin,picroside I,picroside Ⅱ,geniposide and the yield of extract as critical quality attributes( CQAs),the single factor experiment design was used to determine the level of each factor; the Plackett-Burman experiment design was used to select the critical process parameters( CPPs);and the Box-Behnken experiment design was used to optimize the extraction process. The design space of the DGD-4D extraction process was established,and finally,four experimental points were selected to verify the established model. The single factor experiment determined the levels of each factor,including soaking time 60 min and 30 min,water adding volume 12 times and 8 times,extraction time 90 min and 30 min,number of extraction times 3 times and 1 time,as well as extraction temperature 100 ℃ and 90 ℃.By Plackett-Burman experimental design,the DGD-4D water addition,extraction time and number of extraction times were determined to be CPPs. The Box-Behnken experimental variance analysis showed that P of the regression model was less than 0. 01 and the misstated value was more than 0. 01,indicating that the model had good predictive ability,and the operation space of CPPs in the DGD-4D extraction process was determined as follows: the amount of water addition was 10-12 times; extraction time 50-80 min; and number of extraction times was 3 times. The design space of DGD-4D extraction process based on the concept of Qb D is conducive to improving the stability of product quality and laying a foundation for the future development of DGD-4D.

Relationship between MAPK1 gene polymorphism and gefitinib hepatotoxicity in NSCLC patients with activating EGFR mutations / 药学学报

The hepatotoxicity of gefitinib is an important factor limiting its clinical application. In order to control the toxicity, we conducted this study to find the gene variation that can explain and predict the occurrence and severity of hepatotoxicity of gefitinib. Ninety patients with non-small cell lung cancer were included in the retrospective clinical study. Detailed hepatotoxicity induced by gefitinib and epidemiological characteristics were recorded. Twenty-six candidate single-nucleotide polymorphisms of molecular targets, metabolic enzymes, transporters and chemokines were genotyped by matrix-assisted laser desorption/ionization time-of-flight platform. Various confounding factors, such as age, gender and smoking status, were included in the follow-up analysis and variability in the extent of hepatotoxicity was best explained by a multivariate logistic regression model incorporating. The severity of hepatotoxicity was associated with mitogen-activated protein kinase 1 rs13515 (OR=9.467, P=0.074). The research about pharmacogenomic of gefitinib identified the determinants of the drug-induced liver injury. These findings provide a basis to design clinical trials targeting a particular toxicity of gefitinib or similarly targeted agents to benefit patients on long-term gefitinib treatment.

Fingerprinting and multi-indicator quantitative analysis of Mongolian drug Digeda-4 decoction / 中国中药杂志

To establish the high performance liquid chromatography (HPLC) fingerprint for Digeda-4 decoction (DGD-4D), determine the contents of aesculetin, geniposide, picroside Ⅰ, picroside Ⅱ and ellagicacid in DGD-4D, and provide the scientific foundation for quality control of DGD-4D. The analysis was performed on Diamonsil(2) C₁₈ (4.6 mm×250 mm,5 μm) column, with methanol-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL·min⁻¹; injection size was 10 μL; temperature was maintained at 30 °C, and the detection wavelength was set at 254 nm. The common mode of DGD-4D HPLC fingerprint was established, and the hidden information was analyzed by Chemometrics. Chromatographic peaks for DGD-4D were identified by HPLC and quantitative analysis was conducted for characteristic peaks. There were 17 common peaks in the fingerprints and the similarity of the fingerprints was over 0.9 in all 15 batches. The samples were broadly divided into four kinds by principal component analysis and clustering analysis. Four marker compounds were verified by partial least squares discriminant analysis, and No. 9, 12 and 14 peaks were identified as geniposide, picroside Ⅱ, and picroside Ⅰ respectively. The average recoveries were in the range of 95.91%-97.31%. The HPLC fingerprint method for content determination is reliable, accurate, rapid, simple, and reproducible, and can be used as one of the effective methods to control the quality of DGD-4D.

Comparison on the preparation of rat models of pneumonia by different infection route / 中国实验动物学报

Objective To explore the preparation of a rat model of pneumonia model induced by Pseudomonas aeruginosa( PA) using different methods,and to lay the foundation for further studies. Methods 48 SD rats were randomly divided into 4 groups:the control group (A), the intratracheal injection group (B), the trachea cannulation group (C) and the intranasal inoculation group ( D) . After intervention with different treatment modalities, the body weight,tempera-ture,white blood cell count and lung pathological changes in the rats of all groups were detected at 5, 10, 15 days. Results 1. The behavior, body weight, temperature, leukocytes and pathological inflammatory changes of the lung in rats of the model groups were significantly different from that of control group. 2. Pseudomonas aeruginosa was detected in rats of all the model groups, but the control group was negative. Conclusions Rat model of Pseudomonas aeruginosa infected pneu?monia can be successfully established by intranasal inoculation. This method can avoid the inflammatory interference from operation, and is simple and suitable for popularization.

Effect of Qingfei Mixture () on pediatric mycoplasma pneumoniae pneumonia with phlegm heat obstructing Fei (Lung) syndrome / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effect and mechanism of Qingfei Mixture (), a Chinese medicine, in treating mycoplasma pneumonia (MP) in MP patients and rat model METHODS: A total of 46 MP children with phlegm heat obstructing Fei (Lung) syndrome were randomly assigned to two groups by the method of random number table, with 23 children in each group. The control group was treated with intravenous infusion of azithromycin; the treatment group received intravenous infusion of azithromycin and oral administration of Qingfei Mixture. The treatment course was 7 days. Major symptoms and minor symptoms were observed and scored before and after treatments. A rat model of MP was also established. A total of 120 wistar rats were randomly divided into 5 groups: a normal group, infection group, Qingfei Mixture treatment group, azithromycin treatment group, and Qingfei Mixture + azithromycin treatment group. Each group contained 24 rats, from which every 6 were euthanatized 1, 3, 7 and 14 days after infection. MP DNA in pulmonary tissue homogenates was detected using real-time fluorescence quantitative polymerase chain reaction. Pathology was assessed after hematoxylin (HE) staining and lung tissue pathology scores were determined in pulmonary tissue. Transmission electron microscopic detection and electronic image analysis were performed on lung tissue 3 days after infection. Interleukin (IL)-17 was detected in serum using enzymelinked immunosorbent assay (ELISA) 7 days after infection.</p><p><b>RESULTS</b>In the clinical study, both control and the treatment group showed improved results on removing symptoms of phlegm heat syndrome compared to the control group (P<0.05). In animal experiments, On the 7th day after MP infection, as detected by electron microscopy, the pulmonary capillary basement membranes of the azithromycin + Qingfei Mixture treatment group were much thinner than those of the azithromycin or Qingfei mixture treatment groups (P<0.05). The level of serum IL-17 in the azithromycin + Qingfei Mixture treatment group was lower than that in the azithromycin or Qingfei Mixture groups (P<0.01).</p><p><b>CONCLUSION</b>Both Qingfei Mixture and azithromycin have therapeutic effects on mycoplasma pneumoniae pneumonia, but the combination of both agents had the greatest effect.</p>

Effects of Cyclin A1 on the Proliferation of SKM-1 Cells and Its Potential Role in Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Cyclin A1 on the proliferation of SKM-1 cells and its underlying role in myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>Cyclin A1 was knocked down with its small interfering RNA (siRNA). The efficiency of siRNA transfection was measured by Western blot and RT-PCR. Then the proliferation of SKM-1 cells and the expression of CDK2,RUNX1 and SRSF2 with and without knockdown of Cyclin A1 recorded and analysed respectively.</p><p><b>RESULTS</b>Cyclin A1 was knocked down by siRNA after transfected for 48 h. The kncokdown of Cyclin A1 inhibited the proliferation of SKM-1 cells and down-regulated the expression of CDK2, RUNX1 and SRSF2, and these effects were at least partially mediated through RUNX1 and SRSF2 signaling pathway.</p><p><b>CONCLUSION</b>Cyclin A1 plays an important role in the proliferation of SKM-1 cells. These findings provide new insights into the pathogenesis of MDS, and it may be a potential target in the treatment of MDS.</p>

The maternity beds demands and gaps under the universal two-child policy / 中国卫生政策研究

With the universal two-child policy implementation, the birth rate increased, posing challenges to the maternity beds resource allocation. Based on the birth rate and the method of health service demand, we calculat-ed the maternity beds demands and gaps during the period from 2016 to 2020 . Results showed that numbers between 73 478 and 99 004 of maternity beds are needed annually and mainly allocated to eastern and central areas as well. In addition, the maternity beds of different delivery institutions should be adjusted and the hierarchical diagnosis system should be improved in order to alleviate the pressure of the obstestric acceepts in big cities like Beijing and Shanghai.

The pediatrician demands and gaps under the universal two-child policy / 中国卫生政策研究

With the universal two-child policy implementation, the birth rate increased, posing challenges to pediatrician allocation. Based on the birth rate, we calculated pediatrician demands and gaps during the period from 2016 to 2020 by the method of health service demand. The results showed that except Beijing and Shanghai, the ped-iatrician supply and demand ratios are less than 0 . 80 and additional numbers of pediatricians ranging from 191 981 to 198 287 are needed to meet the service demands. We recommend increasing the number of pediatricians taking both national supply-demand ratios and gaps by rationally using reasonable enrolment quota and improving the treatment and other reasonable ways to increase pediatricians. In addition, we should enhance information disclosure and guid-ance, and improve the hierarchical hospital visit system to alleviate the pressure of big cities.

Analysis on the premarital healthcare policy in Guangxi province based on the policy framework / 中国卫生政策研究

Based on the policy framework , the premarital healthcare policy of Guangxi was analyzed from four aspects including policy background , process, content and actors.We analyzed the factors influencing this policy and their interactions , and discussed the important impact of Thalassemia prevention , multi-sectoral consensus and appro-priate premarital healthcare measures to the policy building and implementation .Then the lessons such as strengthe-ning the multi-sector cooperation , specifically making clear the pre-marital screening projects and implementing the“one-stop” service model were summed up in Guangxi province .Finally, combined with existing major problems , some policy recommendations were put forward including clarify the funding criteria and strengthen the premarital healthcare publicity in order to promote the sustainable development of premarital healthcare services .

Effects of premarital healthcare services on birth defects:A case study of Guangxi province / 中国卫生政策研究

Objective:To discuss the effects of premarital health care services on birth defects in order to pro-vide some references for promoting the premarital policy .Methods: The rate of premarital healthcare and the inci-dence of birth defects and other relevant data during the period from 2000 to 2014 were collected in Guangxi province using SPSS18 .0 statistical software to carry out descriptive analysis , rank correlation analysis and time series analysis of the relevant collected data .Results: With the increase in premarital healthcare rate , the incidence of birth de-fects, neural tube defects , cleft lip and fetal edema syndrome decreased .Having a premarital subsequent effect , the higher the premarital healthcare rate , the lower of the incidence of the congenital heart disease and neural tube de-fects in the next year .Conclusion:The premarital healthcare services play an important role in reduction of birth de-fects and people's health quality improvement .As a suggestion , more attention should be paid to premarital health-care services and strengthen them , intended to improve people's health and reduce birth defects incidence .

A study on“one-stop”premarital healthcare services based on SWOT analysis / 中国卫生政策研究

Based on SWOT analysis and relevant information about the premarital healthcare services through on-site survey and literature research , this paper systematically analyzed the Strengths , Weaknesses , Opportunities and Threats of “one-stop” premarital healthcare services .It also puts forward some strategic suggestions on premarital healthcare services in China including enhancing communication and cooperation between the related national minis -tries and commissions , building a government-led multi-sectoral coordination mechanism , increasing the resource in-vestment , promoting the premarital health care service and pre-pregnancy physical examination integration , and strengthening publicity and mobilization of “one stop” service for premarital healthcare service policy recommenda-tions to provide a basis for decision-making in the promotion of the policy-related works .

Analysis of the premarital healthcare service policies implementation:Based on the Mitt-Horn Model / 中国卫生政策研究

Objective:This article uses the Mitt-Horn Model to analyze the premarital healthcare policies imple-mentation, in order to seek viable path for the effective implementation .Methods: Eight provinces, namely Guangxi, Fujian, Yunnan, Jiangsu, Hubei, Heilongjiang, Tianjin and Beijing were chosen to carry out the field research .Relevant information and data were collected through interviews , questionnaires, site visits, etc., and all relate to the implementa-tion of a qualitative premarital analysis policy .Results:The research areas have refined the local premarital policy targets under the relevant legal framework , and have improved the policy implementation resources and execution environment . Conclusion:The premarital healthcare effects of measures implemented in the country based on the actual situation is good, but there are still some problems such as poor policy implementation with no in-depth sector, resource investment deficit in the policy , insufficient inter-department coordination and other issues affecting the policy implementation results .

Empirical research on the financing mechanism of premarital healthcare services in China / 中国卫生政策研究

In this paper , we discussed the financing mechanisms of premarital healthcare services from their properties combined with the status quo financing in our country .We then provided the scientific basis and decision-making reference for improving financing mechanism to improve the premarital healthcare system .A literature or doc-umentary and field researches have combed the present situation of the premarital healthcare services financing mech -anisms at home and abroad .Related data were collected in eight research areas , namely Fujian, Guangxi, Yunnan, Hubei, Jiangsu, Heilongjiang, Beijing and Tianjin and the related personnel from health administrative department , civil affairs departments , maternal and child healthcare institutions were interviewed in terms of Field Research .From the main content at present , premarital healthcare services belong to the category of quasi-public goods .We dis-cussed the financing mechanisms from the financing levels , sources of funds , fund allocation , payment and financial regulatory and other aspects .Finally, some policy recommendations were put forward including making clear the na-ture of premarital healthcare service property , establishment of a special financial investment mechanism , unify the basic financing service standard after scientific feasibility studies , timely and fully allocate special premarital health-care service funds to strengthen the fund supervision policy recommendations .

Experiences and implications of premarital healthcare services in foreign countries / 中国卫生政策研究

As one of the primary prevention measures to reduce birth defects , the premarital healthcare service has played an important role in the prevention of communicable diseases , maintaining the harmonious and happy mar-riage, health and other aspects of protection of the next generation .Different countries and regions also design and implement the premarital medical examination system based on their actual situations .This article summarized the practices, characteristics and experiences of premarital healthcare services in some foreign countries and regions through several aspects including model , set projects content , financing and other aspects of the application of the premarital results.Finally, this paper put forwards suggestions on the premarital healthcare services in our country to help healthy the carrying out of the premarital work .