Establishment and validation of clinical prediction model for steroid-resistant nephrotic syndrome in children / 中华儿科杂志

Objective: To identify the clinically relevant factors of steroid-resistant nephrotic syndrome (SSNS) in children and establish a predictive model followed by verifying its feasibility. Methods: A retrospective analysis was performed in a total of 111 children with nephrotic syndrome admitted to Children's Hospital of ShanXi from January 2016 to December 2021. The clinical data of general conditions, manifestations, laboratory tests, treatment, and prognosis were collected. According to the steroid response, patients were divided into SSNS and steroid resistant nephrotic syndrome (SRNS) group. Single factor Logistic regression analysis was used for comparison between the 2 groups, and variables with statistically significant differences were included in multivariate Logistic regression analysis. The multivariate Logistic regression analysis was used to identify the related variables of children with SRNS. The area under the receiver operating characteristic curve (ROC), the calibration curve and the clinical decision curve were used to evaluate its effectiveness of the variables. Results: Totally 111 children with nephrotic syndrome was composed of 66 boys and 45 girls, aged 3.2 (2.0, 6.6) years. There were 65 patients in the SSNS group and 46 in the SRNS group.Univariate Logistic regression analysis showed that the 6 variables, including erythrocyte sedimentation rate, 25-hydroxyvitamin D, suppressor T cells, D-dimer, fibrin degradation products, β2-microglobulin, had statistically significant differences between SSNS and SRNS groups (85 (52, 104) vs. 105 (85, 120) mm/1 h, 18 (12, 39) vs. 16 (12, 25) nmol/L, 0.23 (0.19, 0.27) vs. 0.25 (0.20, 0.31), 0.7 (0.6, 1.1) vs. 1.1 (0.9, 1.7) g/L, 3.1 (2.3, 4.1) vs. 3.3 (2.7, 5.8) g/L, 2.3 (1.9,2.8) vs. 3.0 (2.5, 3.7) g/L, χ2=3.73, -2.42, 2.24, 3.38, 2.24,3.93,all P<0.05), were included in the multivariate Logistic regression analysis. Finally, we found that 4 variables including erythrocyte sedimentation rate, suppressor T cells, D-dimer and β2-microglobulin (OR=1.02, 1.12, 25.61, 3.38, 95%CI 1.00-1.04, 1.03-1.22, 1.92-341.04, 1.65-6.94, all P<0.05) had significant correlation with SRNS. The optimal prediction model was selected. The ROC curve cut-off=0.38, with the sensitivity of 0.83, the specificity of 0.77 and area under curve of 0.87. The calibration curve showed that the predicted probability of SRNS group occurrence was in good agreement with the actual occurrence probability, χ2=9.12, P=0.426. The clinical decision curve showed good clinical applicability. The net benefit is up to 0.2. Make the nomogram. Conclusions: The prediction model based on the 4 identified risk factors including erythrocyte sedimentation rate, suppressor T cells, D-dimer and β2-microglobulin was suitable for the early diagnosis and prediction of SRNS in children. The prediction effect was promising in clinical application.

Discovery of novel covalent selective estrogen receptor degraders against endocrine-resistant breast cancer

Endocrine-resistance remains a major challenge in estrogen receptor α positive (ERα+) breast cancer (BC) treatment and constitutively active somatic mutations in ERα are a common mechanism. There is an urgent need to develop novel drugs with new mode of mechanism to fight endocrine-resistance. Given aberrant ERα activity, we herein report the identification of novel covalent selective estrogen receptor degraders (cSERDs) possessing the advantages of both covalent and degradation strategies. A highly potent cSERD 29c was identified with superior anti-proliferative activity than fulvestrant against a panel of ERα+ breast cancer cell lines including mutant ERα. Crystal structure of ERα‒ 29c complex alongside intact mass spectrometry revealed that 29c disrupted ERα protein homeostasis through covalent targeting C530 and strong hydrophobic interaction collied on H11, thus enforcing a unique antagonist conformation and driving the ERα degradation. These significant effects of the cSERD on ERα homeostasis, unlike typical ERα degraders that occur directly via long side chains perturbing the morphology of H12, demonstrating a distinct mechanism of action (MoA). In vivo, 29c showed potent antitumor activity in MCF-7 tumor xenograft models and low toxicity. This proof-of-principle study verifies that novel cSERDs offering new opportunities for the development of innovative therapies for endocrine-resistant BC.

Dynein axonemal heavy chain 10 deficiency causes primary ciliary dyskinesia in humans and mice / 医学前沿

Primary ciliary dyskinesia (PCD) is a congenital, motile ciliopathy with pleiotropic symptoms. Although nearly 50 causative genes have been identified, they only account for approximately 70% of definitive PCD cases. Dynein axonemal heavy chain 10 (DNAH10) encodes a subunit of the inner arm dynein heavy chain in motile cilia and sperm flagella. Based on the common axoneme structure of motile cilia and sperm flagella, DNAH10 variants are likely to cause PCD. Using exome sequencing, we identified a novel DNAH10 homozygous variant (c.589C > T, p.R197W) in a patient with PCD from a consanguineous family. The patient manifested sinusitis, bronchiectasis, situs inversus, and asthenoteratozoospermia. Immunostaining analysis showed the absence of DNAH10 and DNALI1 in the respiratory cilia, and transmission electron microscopy revealed strikingly disordered axoneme 9+2 architecture and inner dynein arm defects in the respiratory cilia and sperm flagella. Subsequently, animal models of Dnah10-knockin mice harboring missense variants and Dnah10-knockout mice recapitulated the phenotypes of PCD, including chronic respiratory infection, male infertility, and hydrocephalus. To the best of our knowledge, this study is the first to report DNAH10 deficiency related to PCD in human and mouse models, which suggests that DNAH10 recessive mutation is causative of PCD.

Effect of Tripterygium wilfordii Polyglycoside on Expression of NFAT2/COX-2 in Kidney Tissues of Rats with Diabetic Nephropathy / 中国实验方剂学杂志

ObjectiveTo explore the underlying mechanism of Tripterygium wilfordii polyglycoside tablets （TWPT） in the prevention and treatment of kidney injury in diabetic nephropathy （DN） through the nuclear factor of activated T-cells 2（NFAT2）/cyclooxygenase-2（COX-2） pathway. MethodForty-two male SD rats of SPF grade were selected and randomly divided into a normal group （n=8） and an experimental group （n=34） after one week of adaptive feeding. The rats in the normal group were fed conventionally. The DN model was established in rats of the experimental group by intraperitoneal injection of streptozotocin （STZ） following one week of feeding on a high-fat and high-glucose diet. After the death and failure cases during modeling were eliminated， the remaining 24 model rats were randomly divided into model group， valsartan （8.33 mg·kg-1·d-1） group， and TWPT （5 mg·kg-1·d-1） group. Rats in normal group and model group were given equal amounts of normal saline by gavage. After six weeks， body weight was measured and urine samples were collected. Blood samples were collected from the abdominal aorta， and then the rats were sacrificed for sampling. Biochemical indicators， such as serum blood urea nitrogen （BUN）， serum creatinine （SCr）， alanine aminotransferase （ALT）， blood lipid， blood glucose， and 24-hour urine total protein （24 h UTP）， were determined. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathology of the kidney. Enzyme-linked immunosorbent assay （ELISA） was used to detect NFAT2 and COX-2 expression levels in the serum. Western blot and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）were adopted to detect NFAT2， COX-2 protein and mRNA expression in kidney tissues， respectively. ResultCompared with the normal group， the model group showed elevated 24 h UTP， BUN， SCr， CHO， TG， and FBG， increased serum NFAT2 and COX-2 production and expression （P<0.01）， and elevated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. In addition， the pathology of the kidney showed enlarged glomeruli， mild proliferation of mesangial cells， and widened mesangial stroma. Compared with the model group， the TWPT group showed decreased 24 h UTP， BUN， SCr， CHO， TG， and FBG （P<0.05，P<0.01）， basically normal glomerular morphology， decreased expression of serum NFAT2 and COX-2 （P<0.01）， and down-regulated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. ConclusionTWPT can alleviate 24 h UTP in DN model rats， protect renal function， and improve renal pathology， and its mechanism of action may be related to the down-regulation of NFAT2/COX-2 expression in the serum and kidney tissues.

The effect of Wendler Glottoplasty to elevate vocal pitch in transgender women / 中华耳鼻咽喉头颈外科杂志

Objective: To evaluate the effect of Wendler Glottoplasty to elevate vocal pitch in transgender women. Methods: The voice parameters of pre-and 3-month post-surgery of 29 transgender women who underwent Wendler Glottoplasty in department of otorhinolaryngology head and neck surgery of Beijing Friendship Hospital from January, 2017 to October, 2020 were retrospectively analyzed. The 29 transgender women ranged in age from 19-47 (27.0±6.3) years old. Subjective evaluation was performed using Transsexual Voice Questionnaire for Male to Female (TVQMtF). Objective parameters included fundamental frequency (F0), highest pitch, lowest pitch, habitual volume, Jitter, Shimmer, maximal phonation time (MPT), noise to harmonic ratio (NHR) and formants frequencies(F1, F2, F3, F4). SPSS 25.0 software was used for statistically analysis. Results: Three months after surgery, the score of TVQMtF was significantly decreased [(89.9±14.7) vs. (50.4±13.6), t=11.49, P<0.001]. The F0 was significantly elevated [(152.7±23.3) Hz vs. (207.7±45.9) Hz, t=-6.03, P<0.001]. Frequencies of F1, F2 and F3 were significantly elevated. No statistical difference was observed in the frequencies of F4. The highest pitch was not significantly altered while the lowest pitch was significantly elevated [(96.8±17.7) Hz vs. (120.0±28.9) Hz, t=-3.71, P=0.001]. Habitual speech volume was significantly increased [(60.0±5.2) dB vs. (63.6±9.6) dB, t=-2.12, P=0.043]. Jitter, Shimmer, NHR and MPT were not obviously altered (P>0.05). Conclusions: Wendler Glottoplasty could notably elevate the vocal pitch, formants frequencies and degree of vocal femininity in transgender women without affecting phonation ability and voice quality. It can be an effective treatment modality for voice feminization.

Association of aggression and social support among adolescents / 中国学校卫生

Objective@#To explore the association between aggression and social support and their gender differences among Chinese adolescence, and to provide a scientific reference for preventing and reducing aggressive behaviors of adolescents.@*Methods@#Conducted a cross sectional survey of 15 623 adolescents in 5 provinces in China, namely, HeiLongjiang, Hubei, Anhui, Guangdong and Yunnan Province. And the Chinese version of the Adolescent Social Support Scale was employed to assess the aggression and social support, life events, psychological characteristics, family condition and demographic characteristics among adolescents.@*Results@#The prevalence of self reported high level of aggression was 23.5%(3 670/15 623). Males reported higher rate of high level aggression than females (24.4% vs 22.5%, χ 2=19.30, P <0.01). Significant association between aggression and social support was identified in univariate analysis ( χ 2=620.68, P <0.01). After controlling for potential confounders, aggression was also significantly negatively associated with social support ( OR =1.27-1.84), and there was dose response relationship between them( P < 0.05 ). Furthermore, the association between aggression and social support was similar among male participants and female participants ( ROR =1.02-1.10, P >0.05).@*Conclusion@#The findings indicate that aggression is associated with social support both in male and female adolescents. Improving the social support for adolescents can reduce their aggressive behaviors.

Construction of an online nomogram of risk factors for postoperative delirium in elderly patients with hip fracture / 中华创伤骨科杂志

Objective:To investigate the risk factors for postoperative delirium in the elderly patients with hip fracture and to construct an online nomogram of the risk factors.Methods:Retrospectively analyzed were the data of 483 elderly patients with hip fracture who had been treated with artificial joint replacement from May 2020 to August 2021 at Department of Orthopaedics (Department of Joint Surgery), Jinling Hospital Affiliated to Medical College of Nanjing University. There were 166 males and 317 females, aged from 61 to 99 years (average, 82.1 years). Fracture types: 333 femoral neck fractures and 150 intertrochanteric fractures. The patients were divided into a delirium group ( n=149) and a delirium-free group ( n=334) according to whether postoperative delirium occurred after surgery. The 2 groups were compared in terms of general data like age, gender, body mass index, and concomitant diseases, as well as in terms of indexes like pre-operative albumin, preoperative hemoglobin, and postoperative C-reactive protein (CRP). Factors with P < 0.05 were included in the multi-factor logistic regression analysis to screen out the risk factors for postoperative delirium. The "rms" package of R software was used to draw the nomogram; the Bootstrap method was used to repeat the sampling 1,000 times for evaluation, calculation of the consistency index ( CI), and drawing of the ROC curve and correction curve; the decision curve was plotted using the "rmda" package. Results:There were significant differences between the delirium group and the delirium-free group in age, preoperative anxiety, Alzheimer＇s disease, history of cerebrovascular disease, preoperative albumin, intraoperative hypotension and postoperative CRP ( P < 0.05). The multifactorial logistic regression analysis showed that high age, preoperative anxiety, Alzheimer＇s disease, preoperative albumin < 35 g/L, and postoperative CRP ≥90 mg/L were the risk factors for postoperative delirium in the elderly patients with hip fracture after artificial joint replacement ( P < 0.05). The area under the ROC curve of the nomogram constructed by incorporating the risk factors for postoperative delirium was 0.894 (95% CI: 0.865 to 0.923) with a CI of 0.889; the calibration curve showed that the calibration curve of this nomogram model tended to be close to the ideal curve. The decision curve analysis showed that the threshold value was 0.01 to 1.00, showing the net benefit rate of this nomogram model > 0 when used to predict the postoperative delirium in the elderly patients with hip fracture. Conclusions:High age, preoperative anxiety, Alzheimer's disease, preoperative albumin < 35 g/L, and postoperative CRP ≥90 mg/L may be the risk factors for postoperative delirium in the elderly patients with hip fracture after artificial joint replacement. The online nomogram based on these factors demonstrates a good value in prediction of postoperative delirium.

Clinical phenotypes of primary ciliary dyskinesia / 中南大学学报(医学版)

Primary ciliary dyskinesia (PCD) is a hereditary disease characterized by airway mucociliary clearance dysfunction. The estimated prevalence of PCD is 1꞉10 000 to 1꞉20 000. The main respiratory manifestations in children are cough, expectoration, chronic rhinitis, sinusitis, and chronic otitis media, while the most common symptoms in adults are chronic sinusitis, bronchiectasis, and infertility. About 50% of patients with certain PCD-related gene variants are combined with situs inversus, and the incidence of congenital heart disease is also high. The pathogenesis behind PCD is that gene variants cause structural or functional disorders of respiratory cilia and motile cilia of other organs, leading to a series of heterogeneous clinical manifestations, which makes it difficult to identify and diagnose PCD. Combining different disease screening tools and understanding the relationship between genotypes and phenotypes may facilitate early diagnosis and treatment for PCD.

The Effects of Decitabine Combined with All-Trans Retinoic Acid on the Number of Immune Cells in Myeloid Neoplasms / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of decitabine (DEC) combined with all-trans retinoic acid (ATRA) on the number of immune cells, efficacy and adverse reactions in the treatment of myeloid neoplasms patients.@*METHODS@#Eighty-four patients with myeloid tumors, including AML, MDS-EB-1 or MDS-EB-2 treated by the regimen containing decitabine in our hospital from January 2009 to October 2019 were enrolled and retrospectively analyzed, among the patients, 21 patients treated with DEC alone, 24 patients treated with DEC combined with ATRA (DEC/ATRA) and 39 patients treated with DEC combined with G-CSF priming regimen (DEC/priming). The changes of peripheral blood immune cell levels before and after treatment of the patients between the three groups were compared, and the differences in clinical efficacy and adverse reactions of the patients between the three groups were also compared.@*RESULTS@#There was no statistical differences in the number of immune cells among the patients in the three groups before treatment (P>0.05). NK cell levels decreased significantly in the patients in DEC and DEC/ATRA group after treatment (P<0.05); After treatment, the levels of CD8+ and CD3+T cells in the patients treated by DEC /priming regimen significantly increased (P<0.05), while the levels of CD3-HLA-DR+ B cells significantly decreased (P<0.05). The overall response rate (ORR) of the patients in DEC/ATRA group (75%) and DEC/priming group (74.36%) was significantly higher than 42.86% in DEC monotherapy group, and the differences showed statistically significant (P<0.05), while the ORR between the patients in DEC/ATRA and DEC/priming group showed no statistic differences (P>0.05). There were no statistical differences in overall survival (OS) and incidence of bleeding between the patients in the three groups (P>0.05). The incidences of grade 3 to 4 bone marrow suppression and the infection rate of the patients in DEC monotherapy and DEC/ATRA group were significantly lower than that in DEC/priming regimen group after treatment (all P<0.05), however, there was no statistical difference between DEC monotherapy and the DEC/ATRA group.@*CONCLUSION@#The efficacy of DEC/ATRA on myeloid neoplasms is comparable to that of DEC/priming regimen, and the anti-myeloid tumor effect of DEC/ATRA regimen may be related to the regulation of NK cells and T cells.

Clinical efficacy of artificial vascular replacement for external iliac artery dissection during renal transplantation / 中华器官移植杂志

Objective:To explore the clinical efficacy of treating external iliac artery dissection in renal transplantation by artificial vascular replacement.Methods:Four sudden intraoperative cases of external iliac artery dissection were selected.After removing vascular sutures, intimal arterial peeling blocked external iliac artery( n=3)and transplanted renal artery( n=1). Artificial vascular replacement of external iliac artery was performed using artificial vessels made from puffed polytetrafluoride ethylene(ePTFE). Secondary perfusion was performed in four transplanted kidneys for anastomosing with internal iliac artery. Results:One patient regained normal renal function within 1 week post-operation.Two cases had delayed graft function.Another case had delayed graft function plus acute rejection.After hemodialysis, renal function normalized at 2-3 weeks post-operation.During a follow-up period of(0.5-5.0)years, transplanted kidney function remained stable, blood supply, skin temperature and movement of operated lower extremities normalized.Conclusions:The incidence of vascular dissection of external iliac artery is not high during renal transplantation.However, the disease has a rapid and dangerous progression.The consequences of delayed intervention are quite serious.Treating external iliac artery dissection with renal transplantation may achieve satisfactory clinical outcomes.

Debridement, antibiotics irrigation and implant retention for acute periprosthetic joint infection after primary total joint arthroplasty / 中华创伤骨科杂志

Objective:To analyze the clinical efficacy of debridement, antibiotics irrigation and implant retention (DAIR) in the treatment of acute periprosthetic infection (PJI) and to explore the risk factors leading to the failure of DAIR.Methods:From January 2010 to January 2021, 122 patients underwent DAIR for acute PJI at Department of Orthopedics, General Hospital of Eastern Theater of PLA. They were 55 males and 67 females, aged from 50 to 86 years (mean, 68.0 years). Their C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), body temperature, white blood cell count and visual analogue scale (VAS) were compared at admission and discharge to analyze the clinical efficacy of DAIR. The 122 patients were assigned into a cured group (81 cases) and an uncured group (41 cases). Risk factors were screened by univariate analysis out of their gender, age, body mass index, site of infection, type of infection (early postoperative infection or acute hematogenous infection), type of surgery (primary or revision), comorbidities, CRP, ESR, albumin and hemoglobin at admission, duration of symptoms, Staphylococcus aureus infection, multiple bacterial mixed infection, and preoperative sinus tract. For the factors of P<0.05, multivariate binary logistic regression analysis was used to determine the risk factors for failure of DAIR. Survival curves were plotted for the patients using DAIR failure as the endpoint event. Results:The CRP, ESR, VAS score, body temperature and white blood cell count at discharge in the 122 patients were significantly lower than the corresponding values at admission ( P<0.05). The success rate of DAIR was 66.39%(81/122). The multivariate binary logistic regression analysis suggested that duration of symptoms over 3 weeks( OR=1.230, 95% CI: 1.092~1.576, P=0.020), Staphylococcus aureus infection( OR=4.607, 95% CI: 2.057~10.318, P<0.001), preoperative sinus tract( OR=6.115, 95% CI: 2.630~14.220, P<0.001) and multiple bacterial mixed infection( OR=2.600, 95% CI: 1.131~5.977, P=0.020) were risk factors for DAIR failure; Kaplan-Meier survival curve also confirmed that the patients with Staphylococcus infection, multiple bacterial mixed infection, duration of symptoms over 3 weeks, or preoperative sinus tract had a significantly lower rate of survival than their controls ( P<0.05). Conclusions:For acute PJI, DAIR can be used to retain the prosthesis and control infection. However, DAIR is not recommended for the patients with Staphylococcus aureus infection, multiple bacterial mixed infection, symptoms lasting more than 3 weeks, or preoperative sinus formation.

SARS -CoV -2 Omicron BA.5 infection recurrence after Paxlovid treatment: The first case report in Hunan Province / 中南大学学报(医学版)

We report a case of coronavirus disease 2019 (COVID-19) patient who was cured by oral administration of nirmatrelvir/ritonavir (Paxlovid). The patient was treated with Paxlovid after being first infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron BA.5 variant. On the 11th day after SARS-CoV-2 nucleic acid test turned negative, SARS-CoV-2 nucleic acid test was positive again, and the threshold of nucleic acid cycle number was equivalent to that of the first infection. The results of two whole gene sequencing showed that it was the same virus strain infection, suggesting that the case was re-positive. Without specific treatment, SARS-CoV-2 nucleic acid detection in nasopharyngeal swab turned negative. It is not uncommon for Paxlovid to recover after treating COVID-19, and most of the patients can recover without specific treatment. However, it is necessary to further study the mechanism that may lead to the recovery of SARS-CoV-2.

Change in serum IgG antibody during the recovery stage of Omicron variant infection in children: an analysis of 110 cases / 中国当代儿科杂志

OBJECTIVES@#To investigate the serum level of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific RBD IgG antibody (SARS-CoV-2 IgG antibody for short) in children with SARS-CoV-2 Omicron variant infection during the recovery stage, as well as the protective effect of SARS-CoV-2 vaccination against Omicron infection.@*METHODS@#A retrospective analysis was performed on 110 children who were diagnosed with coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 Omicron variant infection in Tianjin of China from January 8 to February 7, 2022. According to the status of vaccination before diagnosis, they were divided into a booster vaccination (3 doses) group with 2 children, a complete vaccination (2 doses) group with 90 children, an incomplete vaccination (1 dose) group with 5 children, and a non-vaccination group with 13 children. The clinical data and IgG level were compared among the 4 groups.@*RESULTS@#The complete vaccination group had a significantly higher age than the non-vaccination group at diagnosis (P<0.05), and there was a significant difference in the route of transmission between the two groups (P<0.05). There were no significant differences among the four groups in sex, clinical classification, and re-positive rate of SARS-CoV-2 nucleic acid detection (P>0.05). All 97 children were vaccinated with inactivated vaccine, among whom 85 children (88%) were vaccinated with BBIBP-CorV Sinopharm vaccine (Beijing Institute of Biological Products, Beijing, China). At 1 month after diagnosis, the booster vaccination group and the complete vaccination group had a significantly higher level of SARS-CoV-2 IgG antibody than the non-vaccination group (P<0.05), and at 2 months after diagnosis, the complete vaccination group had a significantly higher level of SARS-CoV-2 IgG antibody than the non-vaccination group (P<0.05). For the complete vaccination group, the level of SARS-CoV-2 IgG antibody at 2 months after diagnosis was significantly lower than that at 1 month after diagnosis (P<0.05).@*CONCLUSIONS@#Vaccination with inactivated SARS-CoV-2 vaccine has a protective effect against Omicron infection in children. For children vaccinated with 2 doses of the vaccine who experience Omicron infection, there may be a slight reduction in the level of SARS-CoV-2 IgG antibody at 2 months after diagnosis. Citation:Chinese Journal of Contemporary Pediatrics, 2022, 24(7): 736-741.

Identification of 24 Rana Species Including Rana dybowskii and Rana chensinensis Based on COⅠ Sequences / 中国实验方剂学杂志

Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I （<italic>CO</italic>Ⅰ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>， <italic>R. chensinensis</italic>， <italic>R. amurensis</italic>， <italic>R. culaiensi</italic>s， and <italic>R. huanrenesis</italic>， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>，<italic> Odorrana</italic>， <italic>Nidirana</italic>， <italic>Hylarana</italic>， and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>， <italic>R. zhengi</italic>， <italic>R. coreana</italic>， and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.

Pathogeny research of pulmonary infection after renal transplantation based onmetagenomics nextgeneration sequencing / 中华器官移植杂志

Objective:Discuss the etiological characteristics of pulmonary infection after renal transplantation and the diagnostic value of metagenomics nextgeneration sequencing (mNGS) technique.Methods:A total of 40 patients with pulmonary infection who were admitted to the Department of Renal Transplantation of the First Affiliated Hospital of Medical College, Xi'an Jiaotong University from January, 2018 to January, 2021 were selected, and identification of pathogens through routine pathogen detection methods and mNGS. The routine pathogen detection methods included: blood culture, bronchoalveolar lavage fluid (BALF) and sputum culture and smear staining, lung histopathology, antigen detection and PCR, etc. BALF were used to search for pathogens by mNGS. Combined with the results of the two groups to give accurate anti-infection treatment, the clinical data were retrospectively analyzed.Results:Eventually 36 patients were cured and discharged, and 4 patients deaths. In 40 cases of pulmonary infection, the BALF mNGS pathogens detection of BALF was positive in 37 cases and negative in 3 patients, with a detection sensitivity of 92.5%. In addition, there were 15 cases of single pulmonary infection and 22 cases of mixed pulmonary infection, including 8 cases of bacterial infection, 9 cases of viral infection and 20 cases of fungal infection, among which pneumocystis (20/40, 50%) and cytomegalovirus (10/40, 25%) were the most common. In contrast, the positive rate of pathogens by routine detection were only 30% (12/40), and the difference between the two detection methods was statistically significant ( χ2=32.92, P<0.05). The diagnostic rates of mixed pulmonary infection were 55% and 10% respectively, the difference was statistically significant ( χ2=18.46, P<0.05), the single type pulmonary infection was 30% and 20% respectively, the difference was not statistically significant( χ2=2.99, P>0.05). Conclusions:mNGS has more advantages than routine pathogen detection methods in terms of pathogen species and distribution, detection time, sensitivity, mixed infection diagnosis rate and benefit. Using mNGS can be more efficient to find pathogens of pulmonary infection after renal transplantation, take accurate treatment, reduce costs, and improve cure rate, such as worth wide application..

RASSF6 plays a role in suppressing cancer and improving the sensitivity of oxaliplatin in gastric cancer cells / 西安交通大学学报(医学版)

【Objective】 To investigate the effects of RASSF6 gene on gastric cancer cells’ proliferation， autophagy， apoptosis， and sensitivity to oxaliplatin chemotherapy. 【Methods】 Gastric cancer BGC823 cells were cultured in vitro and divided into experimental control group （control group）， RASSF6 overexpression group （Oe group）， RASSF6 interference group， and lentivirus control group according to the expression effect of lentivirus gene. The changes in cell proliferation， cell cycle distribution， cell migration， autophagy， apoptosis and sensitivity to oxaliplatin in each group were detected， and the number of autophagy bodies in each group was detected by electron microscopy. Real-time PCR （qRT-PCR） and Western blotting were used to detect the expression levels of apoptosis- and autophagy-related genes in each group. 【Results】 Studies on the biological behavior of gastric cancer BGC823 cells induced by RASSF6 gene expression showed that compared with the control group， the percentage of G0/G1 phase cells in the Oe group increased， while the percentage of G2 and S phase cells decreased， with statistical significance （P<0.05）. The apoptosis rate was significantly increased （P<0.05）. The cell scratch assay showed that the scratch healing rate was significantly decreased （P<0.05）. Studies on the sensitivity of RASSF6 gene expression to oxaliplatin showed that compared with the drug group （L-OHP group）， the apoptosis rate of Oe+L-OHP group was increased significantly （P<0.05）. In the Oe+L-OHP group， the expression of anti-apoptotic protein Bcl-2 decreased， the expressions of Bax and Caspase-3 were increased； the expression of autophagosomes was increased； the expressions of Beclin-1 and P62 and the ratio of LC3-Ⅱ/LC3 were all increased （P<0.05）. 【Conclusion】 The RASSF6 gene plays a role in suppressing gastric cancer cell BGC823， which can increase the sensitivity to oxaliplatin chemotherapy by promoting apoptosis and autophagy.

Structure-based engineering of PET hydrolase from Ideonella sakaiensis / 生物工程学报

Polyethylene terephthalate (PET) is a synthetic polymer consisting of ester bond-linked terephthalate and ethylene glycol. Tremendous amounts of PET have been produced and majority of them enters terrestrial and marine environment as wastes, posing serious threats to the global ecosystems. In 2016, a PET hydrolase from a PET-assimilating bacterium Ideonalla sakaiensis was reported and termed as IsPETase. This enzyme outperforms other PET-hydrolyzing enzymes in terms of its PET hydrolytic activity at ambient temperature, thus holds a great promise for PET biodegradation. In order to improve IsPETase activity, we conducted structure-based engineering to modify the putative substrate-binding tunnel. Among the several variants to the N233 residue of IsPETase, we discovered that the substitution of N233 with alanine increases its PET hydrolytic activity, which can be further enhanced when combined with a R280A mutation. We also determined the X-ray crystal structure of the IsPETase N233A variant, which shares nearly identical fold to the WT protein, except for an open end of subsite Ⅱ. We hypothesized that the smaller side chain of N233A variant might lead to an extended subsite Ⅱ for PET binding, which subsequently increases the enzymatic activity. Thus, this study provides new clues for further structure-based engineering of PETase.

Association between PTCH1 gene polymorphisms and chronic obstructive pulmonary disease susceptibility in a Chinese Han population: a case-control study / 中华医学杂志(英文版)

BACKGROUND@#Chronic obstructive pulmonary disease (COPD) is a leading cause of morbidity and mortality worldwide. Genome-wide association studies in non-Asian population revealed a link between COPD and mutations in the PTCH1 gene encoding Patched1, a receptor in the Hedgehog signaling pathway important for lung morphogenesis and pulmonary function. The aim of this study was to investigate the association between PTCH1 polymorphisms and the COPD risk in the Chinese Han population.@*METHODS@#We performed a case-control study including 296 patients with COPD and 300 healthy individuals. Single-nucleotide polymorphisms in the PTCH1 gene were identified and genotyped based on the linkage disequilibrium analysis in all participants. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were estimated using logistic regression analysis after adjustment for age, gender, and smoking.@*RESULTS@#In total, 28 single-nucleotide polymorphisms were identified in patients with COPD. Among them, "A" allele of rs28491365 (OR: 1.388, 95% CI: 1.055-1.827, P = 0.018), and "G" alleles of rs10512248 (OR: 1.299, 95% CI: 1.021-1.653, P = 0.033) and rs28705285 (OR: 1.359, 95% CI: 1.024-1.803, P = 0.033; respectively) were significantly associated with an increased COPD risk. Genetic model analysis revealed that the "T/T" genotype of rs34695652 was associated with a decreased COPD risk under the recessive model (OR: 0.490, 95% CI: 0.270-0.880, P = 0.010), whereas rs28504650/rs10512248 haplotype CG was significantly associated with an increased COPD risk after adjustment for age, gender, and smoking status (OR: 6.364, 95% CI: 1.220-33.292, P = 0.028).@*CONCLUSIONS@#The study provides a new insight into the role of PTCH1 polymorphisms in the susceptibility to COPD in the Chinese Han population.

Diagnosis and treatment of Pneumocystis jirovecii pneumonia after renal transplantation: a report of 20 cases / 中华器官移植杂志

Objective:To explore the pathogenesis, diagnosis and treatment of pneumocystis jirovecii pneumoniae (PJP) after renal transplantation.Methods:A total of 20 PJP patients were selected from January 2018 to January 2020. The major symptoms included fever, chest tightness, dyspnea and cough with minimal sputum. Chest computed tomography (CT), laboratory tests and metagenomic next-generation sequencing (mNGS) of bronchoscopic pulmonary alveolar perfusion fluid (BALF) were performed. They received cotrimoxazole (SMZ: 18.75-25 mg/kg + TMP: 3.75-5 mg/kg q6 h) and basic regimen of caspofungin (50-70 mg/d) for 14-21 days. At the same time, bacterial, fungal or viral infections were treated, immunosuppressants were tapered or discontinued, supportive therapy and methylprednisolone, immunoglobulin and continuous renal replacement therapy (CRRT) were provided. Chest CT examination was performed for 5-7 days to evaluate the therapeutic effect and clinical data were retrospectively analyzed.Results:Among them, fever was predominant in 13 cases with an average body temperature of (38.8±0.68) ℃ while chest tightness and dyspnea occurred in 7 cases. Fourteen patients presented hypoxemia, 13 received non-invasive ventilator assisted breathing, 1 received mechanical ventilation via endotracheal intubation and the remainders received mask oxygen therapy. Seventeen patients were cured and discharged from hospital for 12~90 days. Three patients died of respiratory and circulatory failures due to deteriorating pulmonary infection. And 19 patients had elevated renal function initially and basically normalized around 2 weeks.Conclusions:PJP is one of the most serious complications after renal transplantation. Timely diagnosis and targeted treatment are essential. And mNGS examination plays a crucial role in the diagnosis of PJP. The basic anti-infective scheme of compound sulfamethoxazole plus carbophennet is efficacious. At the same time, appropriate tapering or discontinuation of immunosuppressants, supportive therapy and assisted respiration are also important.

Assessing in-situ conservation status of threatened medicinal vascular plants in central China / 中国中药杂志

It is of great significance to carry out scientific investigation of threatened species and assess their in-situ conservation status in order to guide the conservation and management of these species within a region. In this study, we explored and assessed the biodiversity and in-situ conservation status of the threatened medicinal vascular plants(TMVPs) in central China. The results showed that there were 276 TMVPs in central China, including 18 critically endangered(CR), 77 endangered(EN), and 181 vulnerable(VU) species. Of which, 222 TMVPs were distributed in 49 national nature reserves, with an in-situ conservation rate of 80.43%. And the in-situ conservation rate of CR, EN and NR species were 83.33%, 77.92% and 81.22%, respectively. The complementary algorithm was used to select the nine national nature reserves with the highest protection and complementary contribution to the TMVPs. The cumulative protection contribution rate of these nine nature reserves was 81.98%. Fifty-four TMVPs were distributed outside of national nature reserves, while mainly distributed within 10 hotspot counties including Badong county, Hubei province, Sangzhi county, Hunan province, Jianghua Yao Autonomous county, Hunan province and so on. In general, TMVPs are well protected by the national nature reserves in central China. However, there are still some conservation gaps. It is necessary to carry out further field investigation on the species with conservation gaps and identify the gap areas, so as to provide scientific guidance for the optimization of the conservation of TMVPs in this region.