Objective:To observe the clinical efficacy of Yin-Yang balancing acupuncture plus Tuina(Chinese therapeutic massage)for post-stroke depression(PSD). Methods:A total of 72 PSD patients were randomized into an acupuncture-Tuina group and a Western medicine group using the random number table method,with 36 cases in each group.The Western medicine group was treated with fluoxetine hydrochloride capsules.The acupuncture-Tuina group was treated with Yin-Yang balancing acupuncture plus Tuina therapy.Both groups were treated for 4 weeks.After treatment,the clinical efficacy was compared,and the changes in Hamilton depression scale(HAMD)score,National Institute of Health stroke scale(NIHSS)score,serum neurotransmitter levels,and electroencephalogram(EEG)were observed. Results:The total effective rate of the Western medicine group and the acupuncture-Tuina group was 83.3%and 94.4%,respectively.There was a significant difference in the total effective rate between the two groups(P<0.05).After treatment,the serum levels of 5-hydroxytryptamine(5-HT),norepinephrine(NE),and dopamine(DA)were higher than those in the same group before treatment,and the intra-group differences were all statistically significant(P<0.05).The serum levels of 5-HT,NE,and DA in the acupuncture-Tuina group were higher than those in the Western medicine group,and the differences between the groups were statistically significant(P<0.05).After treatment,the scores of HAMD and NIHSS in both groups decreased,and the intra-group differences were statistically significant(P<0.05).The scores in the acupuncture-Tuina group were lower than those in the Western medicine group,and the differences between the two groups were all statistically significant(P<0.05).After treatment,the EEG diffuse wave[(delta+theta)/(alpha+beta)]ratio(DTABR)and delta/beta ratio(DBR)in both groups decreased,and the intra-group differences were statistically significant(P<0.05).The ratios in the acupuncture-Tuina group were lower than those in the Western medicine group,and the differences between the two groups were statistically significant(P<0.05). Conclusion:Yin-Yang balancing acupuncture plus Tuina therapy has a better efficacy than oral fluoxetine hydrochloride capsules in the treatment of PSD,which can increase the levels of serum 5-HT,DA,and NE,and regulate brain waves.

Objective:To construct a new reporter mycobacteriophage, ΦFN, based on a nanoluciferase (Nluc) reporter system, and to analyze its ability to detect drug resistance in Mycobacterium tuberculosis ( Mtb). Methods:A Nluc reporter system controlled by P furAma promoter was constructed and integrated into Mycobacterium smegmatis ( Msm) genome to assess its reporting ability in mycobacteria. Then the P furAma- nluc reporter system was integrated into TM4 mycobacteriophage genome to construct a new phage termed ΦFN. A rapid procedure for detecting drug resistance in mycobacteria using ΦFN was established by adjusting conditions such as drug exposure time and time of infection. The susceptibility of 52 clinical isolates of Mtb with known drug resistance to three first-line anti-tuberculosis drugs were tested in 96-well plates. Results:The recombinant Msm mc 2155 expressing P furAma- nluc repoerter system could generate luminescence signal at a low limit of 100 colony-forming unit (CFU), which was lower than the previously reported nluc system controlled by Pleft promoter. The detection limit of ΦFN for mycobacteria reached 100 CFU within 24 h by luminescent microplate reader. After 48 h of antibiotic exposure and 24 h of phage incubation, no luminescence signal could be detected when susceptible strains were below 10 5 CFU, which could effectively distinguish susceptible strains and rapidly detect drug resistance. The drug susceptibility of 52 clinical isolates of Mtb to rifampin, isoniazid and streptomycin was tested using ΦFN, and the accuracy was 51/52, 48/52 and 49/52, respectively, by using the conventional drug susceptibility test, Lwenstein-Jensen culture medium assay, as reference. Conclusions:The new recombinant luminescent reporter phage, ΦFN, showed high accuracy in detecting the drug susceptibility of Mtb to rifampicin, isoniazid and streptomycin and it only took three days. It is expected to be a new simple assay for the rapid detection of drug susceptibility of Mtb.

Objective To investigate the changes of blood related parameters and coagulation function indicators in patients with funge-mia during hospitalization,and analyze the clinical significance of these changes and their impact on the patients'prognosis.Methods The patients with positive fungus blood culture for the first time and diagnosed as fungal bloodstream infection during December 2018 and October 2022 were retrospectively analyzed.Their blood related parameters and coagulation function indicators at admission and the time of positive blood culture were collected,and the differences of these clinical parameters were analyzed.The patients were divided into the survival group and death group based on their prognosis,and the differences in clinical parameters between the two groups were compared using the Mann-Whitney U test.The Logistic regression analysis was used to analyze the risk factors related to death.Results A total of 87 patients were enrolled in the study.The patients'results of monocyte count,lymphocyte count,and platelet(PLT)at the time of positive blood culture were significantly lower than those at admission(P＜0.001),while the levels of platelet-associated pa-rameters MPV and PDW were significantly increased(P＜0.05).The levels of coagulation function indicators such as PT,APTT,and D-dimer at the time of positive blood culture were significantly higher than those at admission(P＜0.05).Compared with the the surviv-al group,the peripheral blood PLT count in the death group at the time of positive blood culture was significantly reduced(P=0.022),while the PT,APTT and TT levels were significantly increased(P＜0.05).Logistic regression analysis showed that the death of patients was significantly correlated with the lower level of PLT(OR=1.46,95％CI:1.08-1.97,P=0.01).Conclusion The fungal infection in hospitalized patients can lead to a decrease in PLT count and the prolongation of PT and APTT.Low levels of PLT are associated with the patients'prognosis,and monitoring its changes can assist in assessing the patients'conditions and prognosis.

Objective:To explore the mechanisms of prickle planar cell polarity protein 1 (PRICKLE1) involved in the occurrence of skeletal Class Ⅲ malocclusion.Methods:After extracting the genomic DNA of all family members of the skeletal Class Ⅲ malocclusion pedigree with maxillary hypoplasia collected in the Department of Orthodontics at the Affiliated Stomatological Hospital of Nanjing Medical University in October 2021, whole exome sequencing and Sanger sequencing were performed to screen pathogenic genes/mutation sites and validate the mutations. Jaw tissue was collected during the operation of orthognathic patients who were treated in the Department of Oral and Maxillofacial Surgery at the same hospital from October 2021 to December 2022. Following the extraction of human jaw bone marrow mesenchymal stem cells and transfection with overexpressing lentivirus (lentiviruses overexpressing the gene of interest served as the wild group, lentiviruses overexpressing mutation site served as the mutant group) and knockdown lentivirus (divided into knockdown group 1 and 2, with transfection interference negative lentiviruses as the control group). Various assays including real-time fluorescence quantitative PCR (RT-qPCR), Western blotting, proliferation and Transwell assays, alkaline phosphatase staining and alizarin red staining were performed. Construction of zebrafish animal model, morpholino oligonucleotide (MO) were injected to knock down the expression of prickle1a and prickle1b in zebrafish (co-knocking group), and the control group was injected with standardized MO as a reference. Transcriptome sequencing, enrichment analysis and co-expression analysis were performed on the zebrafish craniofacial tissues of the two groups.Results:Two patients of this family carried this mutation PRICKLE1 c.113C>T. The transfection experiments showed that compared with the wild group (relative expression of PRICKLE1 was 21.97±0.60), the relative expression of mutant group (5.05±0.05) was significantly reduced ( P<0.05), and cell proliferation and migration ability significantly enhanced ( P<0.05), and osteogenic differentiation ability was significantly reduced ( P<0.05). Compared with the control group, the proliferation and migration ability of cells in the two knockdown groups were significantly enhanced ( P<0.05), and the osteogenic differentiation ability was significantly reduced ( P<0.05). Zebrafish model experiments showed the width of the ethmoid plate was significantly reduced in the co-knocking group (282.50±61.77, t=5.29, P<0.001) compared with the control group (338.80±24.92). Transcriptome data and enrichment analysis showed that the differentially expressed genes were significantly enriched in the mitogen-activated protein kinase (MAPK) signaling pathway after the simultaneous knockdown of prickle1a and prickle1b in zebrafish. Conclusions:PRICKLE1 c.113C>T mutation might suppress the osteoblastic differentiation ability of jaw bone marrow mesenchymal stem cells by downregulating the MAPK signaling pathway, thereby involving the development of skeletal Class Ⅲ malocclusion.

Objective:To analyze the failure patterns and influencing factors of stereotactic ablative radiotherapy (SABR) for early-stage non-small cell lung cancer (ES-NSCLC).Methods:113 cases of ES-NSCLC treated with SABR from 2012 to 2020 in our hospital were retrospectively analyzed. The failure patterns, recurrence time, recurrence site and influencing factors were analyzed. Kaplan-Meier method was used to calculate the local recurrence rate, regional lymph node recurrence rate and distant metastasis rate. Univariate analysis was performed by Log-rank test, and multivariate analysis was performed by Cox model.Results:The median follow-up time was 58 months (range: 6-108 months), and a total of 45 patients (39.8%) recurred. The median recurrence time was 36 months. Distant metastasis (DM) occurred in 31 patients (27.4%) and DM alone in 24 patients (21.2%). Local recurrence (LR) was developed in 12 patients (10.6%) and LR alone in 7(6.2%). Regional lymph node recurrence (RR) occurred in 11 patients (9.7%) and RR alone in 6 patients (5.3%). LR combined with RR was observed in 1 case (0.9%), LR combined with DM in 3(2.7%), LR combined with RR and DM in 1(0.9%), and RR combined with DM in 3(2.7%). The 1-, 2-, 3-, 4-and 5-year recurrence rates were 5.4%, 16.6%, 27.5%, 44% and 51.2%, respectively. Univariate and multivariate analyses suggested that EGFR mutation was an influencing factor of high recurrence rate.Conclusion:ES-NSCLC patients treated with SABR alone have a high recurrence rate, and DM is the most common mode of failure. Follow-up consolidation therapy is recommended, especially for EGFR mutation-positive NSCLC patients.

Objective:To investigate clinical phenotypes of type Ⅳ hereditary hemochromatosis caused by c. 430A>G heterozygous mutation of SLC40A1 gene and the correlation between genotype and phenotype, exploring ferritin cutoff value for screening.Methods:One case of type Ⅳ hereditary hemochromatosis with c. 430A>G heterozygous mutation in the SLC40A1 gene and 5 generations of their family lineage with a total of 47 members who were seen at the First Affiliated Hospital of Nanjing Medical University in July 2020 were studied for systematic clinical investigation. Thirty-nine surviving individuals were tested for ferritin, liver function, fasting plasma glucose (FPG), and sex hormones, and Sanger sequencing was performed to verify the mutation loci and to map the family tree. Spearman correlation analysis was used to explore the relationship between ferritin and other indicators, and receiver operating characteristic curves were used to calculate the ferritin cutoff value for screening for this genotype of hemochromatosis.Results:Ten patients with c. 430A>G heterozygous mutation in the SLC40A1 gene were identified among 39 family members, and five of them were diagnosed with hemochromatosis, presenting incomplete penetrance. The differences in levels of ferritin, aspartate aminotransferase (AST; both P<0.01) and FPG, as well as incidences of hypogonadotropic hypogonadism and arthritis (all P<0.05) between group of mutation positive and group negative were statistically significant, while the difference in alanine aminotransferase (ALT) was not. Spearman correlation analysis showed that, ferritin levels were significantly associated with ALT ( r=0.903), AST ( r=0.879), FPG ( r=0.782), and the incidences of hypogonadotropic hypogonadism ( r=0.798) and arthritis ( r=0.798; all P<0.01) in those with the c. 430A>G heterozygous mutation in the SLC40A1 gene. The ferritin cutoff value for screening of hereditary hemochromatosis with c. 430A>G heterozygous mutation in the SLC40A1 gene was 1 036.7 μg/L, with a sensitivity and specificity of 100% and 94.3%, respectively. Conclusion:The SLC40A1 gene c. 430A>G heterozygous mutation is closely associated with elevated levels of AST and FPG, increased incidences of hypogonadotropic hypogonadism and arthritis, and the ferritin cutoff value is a useful screening parameter.

Forkhead box protein O1 (FOXO1) has been extensively studied as a tumor suppressor. In oral squamous cell carcinoma, studies have demonstrated that FOXO1 can inhibit tumor cell oxidative stress, stemness and epithelial-mesenchymal transition, and promote tumor cell autophagy and apoptosis. FOXO1 may serve as a potential target for the treatment of oral squamous cell carcinoma.

Objective:To analyze the clinical efficacy of different treatment modalities and prognostic factors of patients with Masaoka-Koga stage Ⅲ thymoma.Methods:Clinical data of patients diagnosed with Masaoka-Koga stage Ⅲ thymoma admitted to Affiliated Cancer Hospital of Zhengzhou University from January 2000 to December 2018 were analyzed retrospectively. A total of 133 patients had complete treatment and follow-up data. Kaplan-Meier method was used to calculate the cumulative survival rate, log-rank method was used to compare the survival between two groups, and Cox regression model was used for multivariate analysis.Results:The median follow-up time was 50 months (3-221 months). The median overall survival (OS) was 51 (3-221) months, and the median disease-free survival (DFS) was 45 (2-221) months. The survival rate in the radical surgery group was better than that in the palliative surgery group. The 5- and 10-year OS rates in radical surgery group were 88.2% and 74.4% respectively, while in palliative surgery group were 51.8% and 32.4% respectively ( P<0.001). The 5- and 10-year DFS rates in radical surgery group were 72.2% and 45.5%, respectively, while in palliative surgery group were 32.3% and 16.1% respectively ( P=0.001). The OS in the surgery combined with radiotherapy group was better than that in the surgery alone group. The 5- and 10-year OS rates in the radical surgery group were 82.8% and 64.2% respectively, while in the palliative surgery group were 55.8% and 50.2% ( P=0.033). There was no significant difference in DFS between two groups ( P=0.176). Multivariate analysis showed that age < 50 years old ( HR=0.264, P=0.001), radical resection ( HR=0.134, P<0.001), surgery combined with radiotherapy ( HR=2.778, P=0.009) were independently associated with better OS. Age < 50 years old ( HR=0.550, P=0.046), radical resection ( HR=0.555, P=0.042), and invasion of single organ ( HR=0.111, P=0.003) were independently associated with better DFS. Conclusions:OS and DFS in patients undergoing radical surgery are significantly better than those in their counterparts treated with palliative surgery, which is the most important factor affecting prognosis. Surgery combined with radiotherapy yields better OS. It is necessary to design a rigorous and reasonable multicenter prospective study to evaluate the efficacy of various treatment modalities and prognostic factors.

Objective:To analyze the invasion characteristics and prognostic factors of patients with Masaoka-Koga stage Ⅲ thymoma.Methods:The tumor invasion characteristics of 179 patients who were diagnosed with Masaoka-Koga stage Ⅲ thymoma and treated in Affiliated Cancer Hospital of Zhengzhou University from January 2000 to June 2018 were analyzed retrospectively. According to the treatment methods, all patients were divided into the radical operation group ( n=94), palliative operation group ( n=39) and simple biopsy group ( n=46). The χ2 test was used to compare the classified variables, Kaplan- Meier method was utilized to calculate the cumulative survival rate, log-rank method was used for group comparison and univariate analysis, and Cox’s regression model was used for multivariate analysis. Results:Mediastinal pleural invasion (86.0%) was the most common site, followed by pericardium (50.8%), great vessel (40.8%) and lung (36.3%). The proportion of macrovascular invasion in the radical operation group was 14.9%, significantly lower than 79.5% and 60.9% in the palliative surgery group and biopsy group (both P<0.001). Multivariate analysis showed that the nature of operation ( P<0.001), age ( P=0.011), radiotherapy ( P=0.020) were the independent factors affecting overall survival (OS), while nature of operation ( P<0.001), age ( P=0.004), radiotherapy ( P=0.020), number of invasive organs ( P=0.023) and pathological type ( P=0.016) were the independent factors affecting progress-free survival (PFS). Conclusions:For patients with Masaoka-Koga stage Ⅲ thymoma, mediastinal pleura is the most common site of invasion, pericardium, lung and great vessels are also commonly invaded. The invasion of mediastinal pleura, pericardium and lung exerts slight effect on surgical resectability, whereas great vessel involvement can significantly affect surgical resectability. OS and PFS in patients undergoing radical resection are significantly better than those in patients treated with palliative resection and biopsy. Radical resection is the most important factor affecting prognosis.

Objective:To summarize the experience of radiation oncology residency training programs in western coutries, and provide evidence to improve the present Phase-I Radiation Oncology Residency Training Programs in China.Methods:An electronic questionnaire-based survey was conducted among residents and staffs in 6 top cancer centers in China and 5 centers from Europe and North America to collect their feedback regarding the description and comparison of different programs.Results:A total of 70 responses and 4 papers explaining relevant training programs from 26 residents and 23 teachers in Chinese hopsitals and 20 residents and 1 teacher from Europe and North America were received. The Radiation Oncology Residency Training Programs in China were designed into 2 phases, and the results in the current study were involved with the first phase. Program designs were similar in the following aspects: goal, clinical practice-based training, rotation curriculum, interim and final assessment. However, the total timeframe in the investigated Chinese Phase-I programs was shorter than that in western hospitals (2 to 3 years vs. 4 to 5 years). Chinese programs covered major common diseases, whereas rotation design was performed based on each individual disease in western programs. In Chinese programs, the working hours were mainly 40-60 h every week. Although the working hours were commensurate with local workforce regulations, the residents from MSK program had an outstanding longer working time of 60-80 h every week and treated more patients compared with other programs. Conclusions:The investigated Phase-I Radiation Oncology Residency Training Programs in 6 top cancer centers in China share common features in goals, training modes and assessments with western programs. However, our programs have shorter timeframe and less detailed requirements in individual disease than the western programs.