The main protease (M^pro) of SARS-CoV-2 is an attractive target in anti-COVID-19 therapy for its high conservation and major role in the virus life cycle. The covalent M^pro inhibitor nirmatrelvir (in combination with ritonavir, a pharmacokinetic enhancer) and the non-covalent inhibitor ensitrelvir have shown efficacy in clinical trials and have been approved for therapeutic use. Effective antiviral drugs are needed to fight the pandemic, while non-covalent M^pro inhibitors could be promising alternatives due to their high selectivity and favorable druggability. Numerous non-covalent M^pro inhibitors with desirable properties have been developed based on available crystal structures of M^pro. In this article, we describe medicinal chemistry strategies applied for the discovery and optimization of non-covalent M^pro inhibitors, followed by a general overview and critical analysis of the available information. Prospective viewpoints and insights into current strategies for the development of non-covalent M^pro inhibitors are also discussed.

Objective:To explore the immunological characteristics of peripheral blood and genetic variations of 11 immunodeficiency virus(HIV)-negative children with Talaromyces marneffei(TM) infection, thus enhancing the diagnostic and therapeutic levels of TM infection in children. Methods:Clinical data of 11 HIV-negative children with TM infection who presented to Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from January 2010 to December 2022 were retrospectively analyzed, including clinical characteristics, peripheral immune profile and genetic test results.Results:A total of 11 HIV-negative children with TM infections were recruited, involving 9 males and 2 females with a median age of 19 months.The main clinical manifestations were fever (10/11, 90.91%), cough (10/11, 90.91%) and hepatomegaly (7/11, 63.64%). Common severe complications included acute respiratory distress syndrome (7/11, 63.64%) and septic shock (5/11, 45.45%). Finally, 2 children died.Transient neutropenia occurred in 6 cases (6/11, 54.55%), and lymphocytopenia combined with serum immunoglobulin (Ig) G decrease was observed in 4 cases (4/11, 36.36%). IgA decrease, IgM decrease, IgE decrease, IgM increase and IgE increase were observed in 6 cases, 3 cases, 5 cases, 3 cases, and 2 cases, respectively.Both T-lymphocyte and B-lymphocyte counts decreases was observed in 1 case.Genetic testing was performed in all recruited children, and genetic variations were detected in all of them.Inborn errors of immunity (IEIs) were diagnosed in 8 cases, including 4 diagnosed as CD 40 ligand deficiency with CD40LG variation, 1 of severe combined immunodeficiency with IL2RG variation, 1 of Signal transduction and activator of transcription 3(STAT3)-hyper-IgE syndrome with STAT3 variation and 1 of familial candidiasis type 2 with CARD9 compound heterozygous mutations.In the other 3 cases, 2 carried genetic variations that were likely pathogenic, and 1 case was considered uncertain. Conclusions:The clinical manifestations of HIV-negative children with TM infection are atypical, which is characterized as serious complications and high mortality.Early identification and gene testing to detect potential IEIs can improve the prognosis of TM infection.

Objective:To analyze the clinical features of bronchiectasis in children after severe adenovirus pneumonia and to provide clinical clues for the early diagnosis of bronchiectasis in children after severe adenovirus pneumonia.Methods:A retrospective study was made to analyze the clinical data of 26 children with bronchiectasis after severe adenovirus pneumonia treated in Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from May 2016 to May 2021.Results:A total of 26 cases were reported, including 18 males and 8 females.The median onset age of severe adenovirus pneumonia was 23.0 (15.0, 48.0) months.A total of 23 cases suffered concurrent infections, and bacterial co-infection was the most common (16 cases). High resolution computed tomography (HRCT) showed multiple lobar solids in the lung with/without pleural effusion.During the acute phase, most of the cases were treated with intravenous immunoglobulin (21 cases), mechanical ventilation (20 cases), and systemic glucocorticoids (19 cases). The median age at diagnosis of bronchiectasis was 29.5 (21.0, 56.8) months, and the median time that the patients took to develop into acute adenovirus pneumonia was 6.0 (3.3, 13.0) months.Six cases suffered bronchiectasis alone, and 20 cases had bronchiectasis combined with post-infectious bronchiolitis obliterans (PIBO). Of these 20 cases, 3 cases developed bronchiectasis and PIBO simultaneously, and the remaining 17 cases developed bronchiectasis after PIBO.In the included 26 cases, diffuse bronchiectasis predominated (24 cases), most frequently involving the left lower lobes (21 cases) and right lower lobes (21 cases). Cylindrical bronchiectasis was the most common type (23 cases). All the patients had recurrent cough and wheezing during follow-up, and only 3 cases coughed up pus sputum without hemoptysis.All children had acute exacerbations, which were mostly caused by bacteria (21 cases). Nineteen cases combined with PIBO and 1 case with only bronchiectasis were rehospitalized.There was no cases of surgical resection or death.Conclusions:Bronchiectasis after severe adenovirus pneumonia mostly occurs in patients with or without PIBO.Multiple lobe involvement and co-infection may be a risk factor for PIBO patients to develop bronchiectasis.The clinical manifestations are mostly recurrent cough and wheezing, while sputum and hemoptysis are less common.Pediatricians should promptly perform chest HRCT for early diagnosis of the disease.

Objective:To analyze the clinical and imaging characteristics of pulmonary artery sling (PAS) in children and to explore its diagnosis and treatment strategies and risk factors.Method:s A retrospective study was performed in 98 pediatric patients with PAS who were admitted to Guangzhou Women and Children′s Medical Center, Guangzhou Medical University, from January 2011 to October 2018.All their clinical features, imaging examination, treatment and prognosis were reviewed and analyzed.Result:s (1) Among the 98 patients with PAS, 56 cases were male and 42 cases were female.The age range of onset was 0-19 months with the median age of 2 months, and the age range of diagnosis was 0-84 months with the median age of 4 months.(2) The main clinical manifestations were cough (61/98 cases, 62.24%), recurrent wheezing(47/98 cases, 47.96%), shortness of breath (43/98 cases, 43.88%), cyanosis (19/98 cases, 19.39%), and laryngeal stridor (8/98 cases, 8.16%) and so on.(3) Ninety-six patients underwent cardiac ultrasonography, and the diagnostic rate was 87.50%(84/96 cases), and 62 cases (62/96 cases, 64.58%) of them were found to be associated with other congenital cardiovascular anomalies, among which atrial septal defect was the most common (32/96 cases, 33.33%). (4) Chest CT and three-dimensional reconstruction were performed in 92 PAS children, and the diagnostic rate was 100.00%.Airway stenosis was mainly in the middle and lower sections of the main bronchus (76/92 cases, 82.61%), and bronchobridge was found in 13 cases (13/92 cases, 39.13%). Bronchoscopy was performed in 77 children with PAS, showing complete tracheal cartilage ring in 43 cases (43/77 cases, 55.84%), and tracheobronchomalacia in 13 cases (13/77 cases, 16.88%). (5) Among 67 cases who had received left pulmonary artery (LPA) reimplantation, 5 cases died after operation because of airway stenosis and weaning failure (LPA reconstruction was performed in 1 case, and both LPA reconstruction and tracheal intervention were performed in the other 4 cases), 24 cases lost to follow-up, 38 cases survived, and the respiratory symptoms were improved in 2 to 96 months of follow-up.There were 31 cases of non-surgical treatment, of which 18 cases died of respiratory failure, 7 cases lost to follow-up, and the rest 6 cases showed respiratory symptoms of different degrees in the follow-up period.(6) Multivariate regression analysis showed that conservative treatment was an independent risk factor affecting the prognosis of PAS ( OR=7.45, 95% CI: 1.23-48.68). Conclusions:The combination of cardiac ultrasound, chest CT and bronchoscopy is important to the diagnosis of PAS.LPA reconstruction is the main method to treat PAS, which can improve respiratory symptoms, but its fusion with tracheal intervention poses a high risk of death.Conservative treatment is an independent risk factor that affects the prognosis of PAS children.

Objective:To optimize the water extraction and alcohol precipitation technology of Xuanfei Zhike granule .Methods:Orthogonal test was used to investigate the effects of adding water , decocting time and boiling time on the water extraction , and the effects of relative density , alcohol precipitation concentration and alcohol precipitation time on the alcohol precipitation technology by taking comprehensive score including the amount of hesperidin , the amount of tectoridin and the yield of dry cream as the indices .Re-sults:The preferred water extraction technology was as follows: added 10 times water and extracted 1.5 h firstly, and then added 8 times water and extracted twice with 0.5 h for each.The preferred alcohol precipitation technology was as follows:concentrated the wa-ter extraction to a relative density of 1.05 (measured at 60℃), slowly added 95％ethanol to 80％alcohol solution and stored 18 h at low temperature .Conclusion:The optimal water extraction and alcohol precipitation technology is stable and feasible , which can pro-vide reference for the standardized production of Xuanfei Zhike granule .

Objective Four different methods were examined to identify a safer and more reliable method for tail vein punctures in C57BL/6 mice. Methods In total,320 mice were randomly divided into four groups: a blank group, incandescent lamp baking method group,three-line method group,and combined method group. Blood samples were taken from the left or right peripheral vein of puncture mice. Puncture success rate of each group was recorded. SPSS 13.0 software was used to compare statistical difference among groups. Results Compared with the blank group,success rates of the other three methods were significantly higher(P < 0.001). Further, the three-line method was better than the incandescent lamp baking method(P< 0.001). The success rate of the combined method was significantly higher than the three-line and incandescent lamp baking methods(P< 0.001). Conclusions The combined method greatly improved the success rate of tail vein punctures in C57BL/6 mice. This method is more reliable and should be more widely used in the future.

Objective To investigate the osteogenic differentiation potency of ligament cells in thoracic ossification of the ligamentum flavum (TOLF) and analyze further by using transcriptome high-throughput sequencing.Methods Clinically,the patients with non-TOLF and TOLF (n=10 in each group) who underwent surgery in our hospital from October 2015 to April 2016 were included in this study.The primary ligament cells that derived from the two groups were separately cultured and induced osteogenesis with 15％ strength of cyclic mechanical stress for 12h and 24h using a device called Flexcell FX-4000.The ALP activity was determined to evaluate the osteogenesis using quantitative analysis and ALP staining assay.Real-time PCR and westernblotting were used to detect the mRNA and protein expression of osteogenic-related genes including ALP,BMP-2 and Osteocalcin.Then,three patients in each group were included in the study of transcriptome high-throughput sequencing and bioinformatics analysis using Illumina HiSeqTM 2500 sequencing platform to compare further.Results The morphology of the cells that derived from two groups was basically similar,all presented an elongate spindle-shape.To evaluate the ostogenesis,ALP activity assays including quantitative and staining assays were performed.Under microscope,the ALP staining in the TOLF group was higher than non-TOLF group and increased with the longer duration of stress induction.The result of semi-quantitative analysis showed the stained area and positive cells in TOLF group were more than non-TOLF group significantly at 0 h,and were increased with the induction.The results of quantitative analysis showed ALP activity in the TOLF group was significantly higher than non-TOLF group and were increased with the induction significantly all the time.But no significant change in ALP staining or quantitative analysis was found in non-TOLF.The results of real-time PCR indicated that the expression of ostegenic markers above in the TOLF group was more than non-TOLF group significantly except the expression of OCN at 0 h.The expression of the three ostegenic markers in TOLF group was increased with the stress induction for 12 h and 24 h significantly except the expression of BMP-2 and OCN at 12 h.The results of western-blotting indicated that the expression of the three ostegenic markers above in the TOLF group was more than non-TOLF group significantly except the expression of ALP at 0 h.The expression of the three ostegenic markers in TOLF group was increased with the stress induction for 12 h and 24 h,but only the expression of ALP at 24 h was significant.And no significant change in the expression of mRNA and protein was found in non-TOLF group.In the transcriptome analysis,671 genes of TOLF group were up-regulated and 314 genes were found to be down-regulated compared to the control group.In addition,22 significant GO terms associated with upregulated genes were found to be closely related to ossification.Conclusion TOLF ligament cells have high osteogenic differentiation potency,which could express obvious osteogenesis-related gene spectrum,and differentially expressed genes including L1RL1 、PTHLH、DKK1 、BMP6、SPP1 and FGF1 may be related with the osteogenic potency of ligament cells in thoracic ossification.

Objective To compare the effect of breast conserving surgery and modified radical mastectomy on the clinical efficacy of patients with early breast cancer.Methods CNKI,Chinese Journal Full-text Database (CJFD),China Biology Medicine Disc (CBMdisc),China Dissertation Database (Chinese Dissertation Database,CDDB) google academic and other databases were comprehensive searched.And then the search time was limited to between January 1,2015 and September 1,2017.Key words can be locked for breast conserving surgery,improved surgery for breast surgery,case-control studies,etc.,and then meet the conditions of the literature into the study,for a retrospective analysis.The authors reviewed the literature independently,extracted data and evaluated the risk of bias,and used Review Manager 5.3 software for systematic analysis.Results A total of 1 093 patients with early breast cancer were enrolled in the study.The Meta-analysis showed:there was a significant difference in operation time between the two groups(MD =-30.71,95％ CI:-31.96--29.46,P ＜ 0.01);there was a significant difference in intraoperative blood loss between the two groups(MD =-53.30,95％ CI:-55.38--51.22,P ＜ 0.01);there was a significant difference in postoperative hospital stay (MD =-5.66,95％CI:-7.17--5.17,P ＜0.01) and the incidence of complications (OR =0.30,95％ CI:0.19-0.47,P ＜ 0.01)compared with modified radical mastectomy in early breast cancer patients.There was no significant difference between the two groups in the postoperative recurrence and metastasis (OR =0.78,95％ CI:0.54-1.13,P =0.19).Conclusions In the choice of surgical methods,breast-conserving therapy is better than modified radical surgery,and postoperative recurrence and metastasis rate has no significant difference.

Objective To analyze the clinical and anatomic characteristics of bridging bronchus (BB) malformation in children,and to explore its diagnostic strategy,treatment and risk factors.Methods A retrospective study was performed in 23 pediatric patients with BB who were admitted to Guangzhou Women and Children's Medical Center form May 2010 to October 2016.All their clinical features,imaging examination,treatment and prognosis were reviewed and analyzed.Results (1) Among the 23 patients with BB,15 were males and 8 were females.The age range of onset was 0 to 4 years old with a median age of 3.17 months.(2)The main clinical manifestations were cough (23/23 cases,100.0％),recurrent wheezing (20/23 cases,87.0％),cyanosis (8/23 cases,34.8％) and feeding diffficulty/slow body weight growth (6/23 cases,24.6％).(3) There were 17 cases of type Ⅰ and 4 cases of type Ⅱ according to Wells,and the other 2 cases were anterior BB.There were 2 cases with atypical anatomical morphology in type Ⅰ and type Ⅱ respectively.(4) Twenty-one cases were associated with bronchial stenosis in which the origin of left main bronchus complicated with BB stenosis was the most common(5 cases).Bronchoscopy revealed bronchomalacia in 5 of the 16 patients.Sixteen patients associated with cardiac or vascular anomalies,11 of whom were compound anomalies,and the most common type was sling left pulmonary artery (SLPA) (12 cases).(5) Nine of 12 patients with SLPA received surgical management,ages ranging from 1 month to 8 years old.The respiratory symptoms remitted gradually or disappeared from 4 to 17 months after surgical management.Two died and one has been lost to follow-up among the remaining 3 patients with SLPA.Conclusions BB is a rare tracheobronchial malformation that is often associated with stenosis or bronchomalacia,and cardiac or vascular anomalies.The severity of the disease mainly depends on whether there is airway stenosis or malacia and its extent.The definitive diagnosis of BB requires the combination of different imaging modalities.The combination with simple SLPA could get better curative effect.

Objective To assess the efficacy and safety of combining rociverine tablets with Suoquan cap-sules in the treatment of female patients with overactive bladder(OAB).Methods 152 female patients with OAB were randomly divided into three groups.Patients in rociverine group were administered rociverine tablets 10mg three times a day for 4 weeks.Patients in Suoquan capsule group were administered Suoquan capsules 1.8g three times a day for 4 weeks.Patients in combination group were administered rociverine tablets 10mg and Suoquan capsule 1.8g three times a day for 4 weeks.Each group of patients had to complete 3d voiding diary before and after treatment to record daily frequency of micturition,urgency,urgency incontinence,nocturia and avoiding volumes,OABSS score and drug side effects were also observed.All the data were analyzed to assess the efficacy and safety of treatment.Results There were significant differences in daily frequency of micturition,urgency,urgency incontinence,nocturia,avoiding volumes and OABSS score of patients in each group before and after treatment(all P <0.01 ).Moreover,compared with combination group,rociverine and Suoquan capsule group all had significant differences(F =41.765,53.834, 25.304,37.867,214.791,73.486,all P <0.01 ).There was no significant differences between rociverine and Suoquan capsule group(all P >0.05).The incidence rates of adverse events of rociverine and combination group were 7.8% and 22.6%,respectively,there was significant difference between the two groups(χ2 =4.372,P =0.032). Conclusion Combining rociverine tablets with Suoquan capsules in the treatment of female patients with OAB was effective and safe.