Objective:To explore the differences in bacterial communities within tissues during the process of oral mucosal carcinogenesis, and analyze the relationship between the high-abundance species Prevotella intermedia (Pi) and the occurrence and development of oral mucosal carcinogenesis. Methods:Fresh tissue samples were collected from patients diagnosed with oral leukoplakia (OLK), oral squamous cell carcinoma (OSCC), and healthy controls (HC) at Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, from January 2022 to November 2024, following strict inclusion criteria. Bacterial DNA was extracted from these specimens, and the 2bRAD sequencing for microbiome (2bRAD-M) was employed to analyze and compare the α and β diversity, as well as the community composition of bacteria within tissues, aiming to identify specifically expressed bacteria. Subsequently, paraffin-embedded clinical specimens were collected: 15 cases in the OLK group (including 4 cases of simple hyperplasia, 6 cases of mild dysplasia, and 5 cases of moderate to severe dysplasia), 12 cases in the OSCC group, and 5 cases in the HC group. A 4-nitroquinoline N-oxide (4NQO)-induced OLK progression mouse model was also constructed. Mice were randomly divided into three groups using a random number table, with six in each group. The negative control group was given distilled water to drink; Group 1 was given distilled water containing 4NQO to drink until week 12, while Group 2 was given distilled water containing 4NQO to drink until week 22. After the mice were sacrificed, their tongue tissue were collected and fixed. Fluorescence in situ hybridization (FISH) with specific probes was used to validate the presence of Pi in human and mouse tissue sections, analyzing the correlation between histopathological grading and the invasion depth of Pi.Results:The 2bRAD-M microbial analysis revealed that the relative abundance of Pi in OSCC tissues (10.80%) was significantly higher than in the HC group (0.50%) ( P=0.001) and OLK group (0.70%) ( P=0.002). FISH probe detection showed that the fluorescence intensity of Pi in human OSCC tissues [123.50 (101.00, 142.30)] was higher than in the HC group [0.00 (0.00, 28.50)], simple hyperplasia OLK [0.00 (0.00, 35.25)], and mild dysplasia OLK [24.50 (0.00, 55.50)] groups, with statistically significant differences respectively ( P=0.002, P=0.003, P=0.005). However, there was no significant difference compared to moderate to severe dysplasia OLK [56.00 (28.00, 62.50)] ( P=0.210). The fluorescence area of Pi in human OSCC tissues [8 615.00 (7 439.00, 11 084.00)] was significantly larger than in the HC group [0.00 (0. 00, 45.00)], simple hyperplasia OLK group [0.00 (0.00, 81.00)], mild dysplasia [49.00 (0.00, 151.00)], and moderate to severe dysplasia groups [1 450.00 (454.00, 2 892.00)], with highly significant differences ( P<0.001). There was a significant correlation between the invasive depth of Pi and the degree of histopathological grading ( P<0.001). In mice, the fluorescence intensity of Pi in OSCC tissues [120.00 (110.00, 127.00)] was significantly higher than in the HC group [0.00 (0.00, 12.25)] ( P<0.01), but showed no significant difference compared with the OLK group [50.00 (0.00, 58.00)] ( P>0.05). The fluorescence area of Pi in mice OSCC tissues [11 020.00 (6 790.00, 12 102.00)] was significantly larger than in the HC group [0.00 (0.00, 56.75)] and the OLK group [0.00 (0.00, 751.50)] ( P=0.006, P=0.043). There is a significant correlation between the depth of invasion of Pi and the degree of histopathological grading ( P<0.01). Conclusions:This study suggests that Pi in oral mucosal tissue may be a potential biomarker for early detection of OSCC and play an important role in the carcinogenesis process of oral mucosa.

Objective:To explore the differences in bacterial communities within tissues during the process of oral mucosal carcinogenesis, and analyze the relationship between the high-abundance species Prevotella intermedia (Pi) and the occurrence and development of oral mucosal carcinogenesis. Methods:Fresh tissue samples were collected from patients diagnosed with oral leukoplakia (OLK), oral squamous cell carcinoma (OSCC), and healthy controls (HC) at Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, from January 2022 to November 2024, following strict inclusion criteria. Bacterial DNA was extracted from these specimens, and the 2bRAD sequencing for microbiome (2bRAD-M) was employed to analyze and compare the α and β diversity, as well as the community composition of bacteria within tissues, aiming to identify specifically expressed bacteria. Subsequently, paraffin-embedded clinical specimens were collected: 15 cases in the OLK group (including 4 cases of simple hyperplasia, 6 cases of mild dysplasia, and 5 cases of moderate to severe dysplasia), 12 cases in the OSCC group, and 5 cases in the HC group. A 4-nitroquinoline N-oxide (4NQO)-induced OLK progression mouse model was also constructed. Mice were randomly divided into three groups using a random number table, with six in each group. The negative control group was given distilled water to drink; Group 1 was given distilled water containing 4NQO to drink until week 12, while Group 2 was given distilled water containing 4NQO to drink until week 22. After the mice were sacrificed, their tongue tissue were collected and fixed. Fluorescence in situ hybridization (FISH) with specific probes was used to validate the presence of Pi in human and mouse tissue sections, analyzing the correlation between histopathological grading and the invasion depth of Pi.Results:The 2bRAD-M microbial analysis revealed that the relative abundance of Pi in OSCC tissues (10.80%) was significantly higher than in the HC group (0.50%) ( P=0.001) and OLK group (0.70%) ( P=0.002). FISH probe detection showed that the fluorescence intensity of Pi in human OSCC tissues [123.50 (101.00, 142.30)] was higher than in the HC group [0.00 (0.00, 28.50)], simple hyperplasia OLK [0.00 (0.00, 35.25)], and mild dysplasia OLK [24.50 (0.00, 55.50)] groups, with statistically significant differences respectively ( P=0.002, P=0.003, P=0.005). However, there was no significant difference compared to moderate to severe dysplasia OLK [56.00 (28.00, 62.50)] ( P=0.210). The fluorescence area of Pi in human OSCC tissues [8 615.00 (7 439.00, 11 084.00)] was significantly larger than in the HC group [0.00 (0. 00, 45.00)], simple hyperplasia OLK group [0.00 (0.00, 81.00)], mild dysplasia [49.00 (0.00, 151.00)], and moderate to severe dysplasia groups [1 450.00 (454.00, 2 892.00)], with highly significant differences ( P<0.001). There was a significant correlation between the invasive depth of Pi and the degree of histopathological grading ( P<0.001). In mice, the fluorescence intensity of Pi in OSCC tissues [120.00 (110.00, 127.00)] was significantly higher than in the HC group [0.00 (0.00, 12.25)] ( P<0.01), but showed no significant difference compared with the OLK group [50.00 (0.00, 58.00)] ( P>0.05). The fluorescence area of Pi in mice OSCC tissues [11 020.00 (6 790.00, 12 102.00)] was significantly larger than in the HC group [0.00 (0.00, 56.75)] and the OLK group [0.00 (0.00, 751.50)] ( P=0.006, P=0.043). There is a significant correlation between the depth of invasion of Pi and the degree of histopathological grading ( P<0.01). Conclusions:This study suggests that Pi in oral mucosal tissue may be a potential biomarker for early detection of OSCC and play an important role in the carcinogenesis process of oral mucosa.

Objective:To explore the role of SIRT3 down-regulation in skeletal muscle injury through the changes of sirtuin 3 (SIRT3) expression in mouse skeletal muscle under iron excess in vitro or in vivo.Methods:Murine preosteoblast myoblast C2C12 cells were incubated in a medium supplemented with ferric ammonium citrate (FAC). The proliferation, apoptotic were assessed, the cell morphology was observed, and the expression of SIRT3 mRNA, protein and activity were detected. ICR mice were randomly divided into control group, FAC group and FAC+ deferrioxamine (deferoxamine, DFO) group. Normal saline was injected in the control group. FAC followed by injection of normal saline in the FAC group; and FAC followed by DFO in the FAC+ DFO group. The non-heme iron level, content of SIRT3 protein and in situ apoptosis were detected, morphology of skeletal muscle was observed.Results:Proliferation of C2C12 cells was inhibited, and the apoptotic rate were increased by FAC ( P<0.05). The mRNA, protein and activity of SIRT3 decreased by FAC ( P<0.05). The cells gradually shrank, and the length and number of myotubes were decreased by FAC. Both control and FAC+ DFO groups showed lower levels of non-heme iron in skeletal muscle compared with FAC group ( P<0.05). The levels of SIRT3 protein were decreased in FAC group compared with control group, while increased in FAC+ DFO group with FAC group ( P<0.05). The apoptotic indexes in control and FAC+ DFO groups were lower than that in FAC group. Compared with the control group, the disordered cell arrangement, fat deposition and inflammatory cell infiltration were presented in FCA group, and the change was alleviated in FAC+ DFO group. Conclusion:Iron excess can lead to the decrease of skeletal muscle mass in mice, and the mechanism may be related to the down-regulation of SIRT3 level.

Objective:To explore the clinical effect of observation and psychological intervention, splint or brace fixation as well as surgical treatment on habitual dislocation of the hip (HDH) and to combine the authors' data with a compilation of the cases from the literature, evaluingating the epidemiological characteristics of HDH and the treatment scheme to maintain the stability of hip joint by systematic literature review in children.Methods:A retrospective analysis of the relevant data of 11 patients (12 hips) with HDH were treated from March 2007 to March 2021, including 2 boys and 9 girls. The age of the first dislocation was 2.25 (1.66, 3.75) years old and 4.33 (3.33, 6.17) years old at the age of diagnosis. At the same time, the relevant literature reports were searched from 1932 to 2022, and 24 HDH patients reports and clinical studies were confirmed to be included in this study according to the inclusion and exclusion criteria. The data of 33 patients (38 hips) with HDH who were obtained in the literature, including 5 boys and 28 girls. The age of the first dislocation was 2.00 (1.50, 2.00) years old and 4.00 (2.55, 5.00) years old at the age of diagnosis. A total of 44 patients (50 hips) with HDH who were analyzed, including gender, age of first dislocation, age of diagnosis, mode of stimulating dislocation, side and direction of dislocation, accompanying symptoms, family history, trauma history and imaging examination. The data of 41 patients with HDH (3 patients were excluded due to lack of treatment description) were treated with observation and psychological intervention in 19 patients, splint or brace fixation in 13 patients, and surgical treatment in 9 patients. The femoral neck-stem angle, acetabular index, central edge angle (CE angle) and Reimers instability index were measured by AutoCAD software, and the hip function was evaluated by Harris standard. Meantime, the disappearance time of dislocation was recorded.Results:The average follow-up time of 44 patients (50 hips) with HDH were 4.05±2.93 years old, and the time of disappearance of dislocation after treatment were 1.28±1.19 years old. Patients were diagnosed as 7 boys and 37 girls, 30 right and 8 left as well as 6 bilateral, the age of the first dislocation was 2.64±1.54 years old and 4.52±2.64 years old at the age of diagnosis. The data of 39 patients took hip flexion, adduction, internal rotation or hip flexion and adduction as the mode of stimulating dislocation. There were 43 patients with posterior dislocation, 41 patients with an audible "click" sound during dislocation, 36 patients with painless dislocation, and 37 patients with "vacuum phenomenon" were captured at the moment of dislocation. All patients with HDH had no specific family history and obvious history of trauma. There was no significant difference in general data between observation and psychological intervention group, splint or brace fixation group as well as surgical treatment group ( P>0.05). Harris standard to evaluate hip function, CE angle and Reimers instability index of the affected side were significantly different from those before and after treatment at the moment of dislocation ( H=127.51, P<0.001; H=55.70, P<0.001; H=54.69, P<0.001). Compared with the immediate disappearance of dislocation in the surgical treatment group, the disappearance time of dislocation in the observation and psychological intervention group and the splint or brace fixation group were significantly longer, and the difference was statistically significant ( H=20.83, P<0.001). Conclusion:Without specific family history and obvious trauma at young girls, painless posterior dislocation of hip with an audible "click" sound and "vacuum phenomenon" at the moment of dislocation are the significant epidemiological characteristics of HDH. Observation and psychological intervention, splint or brace fixation are recommended as the initial treatment scheme. When conservative treatment is ineffective, surgical treatment is needed to stabilize the hip joint quickly.

Objective:Compared with closed reduction and percutaneous pinning (CRPP) treatment, evaluating the clinical observation of utilizing a transolecranon pin joystick technique combined with CRPP in the teatment of multidirectionally unstable supracondylar humeral fractures in children.Methods:From thirty nine pediatric multidirectionally unstable supracondylar humeral fractures hospitalized between January 2012 and January 2019, twenty seven males (69.23%) and twelve females (30.77%) were included in the study, with a average age of 6.68±2.52 years (range, 2.17-13.75 y), twenty three fractures (65.7%) were treated with CRPP (CRPP group) and the remaining Sixteen fractures (41.03%) were treated utilizing a transolecranon pin joystick technique combined with CRPP (joystick group). Both groups were followed over 16 weeks. The paired sample t test or χ2 test and Fisher's exact test were used to compare the surgical time, times of fluoroscopy, quality of reduction and neurological or vascular complications, Baumann angle, carrying angle, lateralcapitellohumeral angle, postoperative range of motion as well as function-al outcomeduringthe Sixteen weeks and the last follow-up appointment. Results:All caseswere followed up for 1.98±1.43 years, and all fractures achieved clinical healing at 4 to 6 weeks postoperation. The surgical time and times of fluoroscopy were significantly shorter for patients in the joystick group (27.17±9.68 min, 24.25±5.92 times) when compared with CRPP group (48.59±15.75 min, 49.65±23.83 times, P<0.05). All cases showed restoration of the normal lateral capitellohumeral angle. Compared with Baumann angle of normal contralateral upper extremity during the sixteen weeks and the last follow-up appointment, the quality of reduction on the anteroposterior radiographic view was significantly better for patients in the joystick group than that of CRPP group ( P<0.05). The Baumann angle of the affected upper extremity was 77.70°±2.16°, and that of the normal contralateral upper extremity was 73.78°±4.04° in the CRPP group, joystick group was 73.06°±1.81° and 72.81°±3.45°, respectively at the sixteen weeks follow-up. The Baumann angle of the affected upper extremity was 77.13°±2.20°, and that of the normal contralateral upper extremity was 74.17°±4.17° in the CRPP group, joystick group was 72.69°±1.70° and 73.38°±3.48°, respectively at the last follow-up. The range of motion and clinical outcomes based on the criteria of Flynn were similar in both groups ( P<0.05). The elbow joint function of excellent and good rate of the criteria of Flynn was 82.61%, elbow flexion was 134.13°±8.61°, elbow extension was -3.48°±6.47° in the CRPP group, joystick group was 81.25%, 132.19°±9.48° and -3.44°± 4.37°, respectively at the sixteen weeks follow-up. The elbow joint function of excellent and good rate of the criteria of Flynn was 91.30%, elbow flexion was 140.14°±5.76°, elbow extension was -0.65±3.79° in the CRPP group, joystick group was 93.75%, 141.88°±5.12° and -0.31°±3.86°, respectivelyat the last follow-up. No immediate postoperative complications were observed. Conclusion:A transolecranon pin joystick technique combined with CRPP is a safe and effective method, can decrease surgical time and times of fluoroscopy and improve quality of reduction with no increasing risk of complications for closed reduction of multidirectionally unstable supracondylar humeral fractures in children.

Objective: To construct influenza B virus Vero cell adapted strain by genetic recombination technology by using the influenza B virus Vero cell adapted strain as the parent strain. Methods: The chick embryo and Vero cell were co-infected with influenza virus Vero cell adapted strain B/Malaysia/2506/2004 Va (Bv) and the vaccine strain B/massachusetts/2/2012 (BX-51B) recommended by WHO. The reassortants were screened with the anti-Bv serum. Plaque-purified reassortants were used to screen for Vero cell-adapted influenza B virus strains containing the surface antigen of the epidemic strain. Results: A Vero cell-adapted influenza B virus strain was obtained with successive passage in Vero cells. The hemagglutination inhibition test and the one-way immunogold agar diffusion test both showed that the reassortant virus was homologous to NYMC BX-51B, and sequence analysis result showed that the reassortment virus has the same HA and NA gene with the vaccine strain. Conclusion B/Malaysia/2506/2004Va (Bv) can be used as a parent strain to prepare Vero cell vaccine against influenza B virus.

Objective To evaluate the immunogenicity and safety of inactivated poliovirus vaccine derived from Sabin strain (sIPV) in Banna mini pigs, and to provide experimental evidence for the new animal model. Methods sIPV vaccines which are listed at Institute of Medical Biology at Chinese Academy of Medical Sciences were used in this study. The groups of intramuscular sIPV and the wild strain IPV injections (IPV derived from wild strain, wIPV) were designed, and the saline group was used as a negative control group. The Banna mini pigs in various groups were immunized at 0, 1 and 2 months. Blood samples were collected before immunization and on days 30 after each immunization. Levels of neutralizing antibodies were tested for evaluating immunogenicity. The safety was evaluated by observation of the status and weight of mini pigs. Results After the three dose immunization schedules in the Banna mini pigs, the seroconversion rates of type Ⅰ,Ⅱ and Ⅲ sIPV experimental groups and wIPV group were all up to 100%. The neutralizing antibody levels in all the three types were much higher than the protective titer 1: 8. The weight of mini pigs increased after vaccination. Conclusions The sIPV vaccine has good immunogenicity and safety in Banna mini pigs. Banna mini pigs could be a new animal model for evaluation of sIPV vaccine.

[Objective] To explore the risk factors for mortality of bloodstream infections in the patients with hematologic diseases,so as to provide evidence for reasonable and effective application of treatments.[Methods] The clinical data of 242 cases of bloodstream infections who were hospitalized from Jan 2012 to Jun 2016 were analyzed retrospectively,then the analysis was performed for risk factors.The statistical analysis was processed by SPSS 19.0.[Results] A total of 266 strains of pathogens were isolated,including 99 strains of gram-positive bacteria,accounting for 37.2％,and 164 strains of gram-negative bacteria,accounting for 61.7％.Multivariate analysis showed that the significant independent risk factors for mortality were active states of hematologic diseases (P =0.007,OR =5.622,95％ CI 1.586 ～ 19.924),presentation with septic shock(P =0.007,OR =4.978,95％ CI 1.560 ～15.884),cardiac insufficiency (P =0.001,OR =11.878,95％ CI 2.760 ～ 51.120),level of albumin less than 35 g/L (P =0.036,OR =3.468,95％ CI 1.087 ～ 11.066),polymicrobial infection (P =0.010,OR =6.024,95％ CI 1.540 ～ 23.563),and Staphylococcus haemolyticus (P =0.001,OR =19.308,95％ CI 3.392 ～ 109.888)/Enterococcus (P =0.002,OR =15.266,95％ CI 2.817 ～82.728) infection.The survival curves show that the inappropriate initial antimicrobial therapy group or presentation with any one of the independent risk factors had a lower probability of survival than the control group.[Conclusions] Bloodstream infections in patients may cause high mortality rate,so it is necessary that we use antibiotic reasonably and spare no effort to reduce the mortality rate by appropriate application of antimicrobial therapy and effective intervention of the risk factors.

Objective To evaluate the correlation of the single nucleotide polymorphisms (SNPs) in CDH13 with non-small cell lung cancer (NSCLC) . Methods 115 patients with NSCLC and 110 healthy controls were included in present study. Two SNPs (rs11646213 and rs7195409) in CDH13 were genotyped using TaqMan method. The association of these two SNPs with NSCLC was calculated and assessed. Results The genotypic and allelic frequencies of rs11646213 showed significant difference between NSCLC patients and the control group (P<0.05), (OR=0.464, 95% CI:0.273~0.789) . The genotypic and allelic frequencies of rs7195409 showed significant difference between the stage I+II and stage III+IV groups (P<0.05), (OR=0.491, 95% CI:0.243~0.991) . Conclusions The rs16146213 has a strong association with NSCLC and G allelic showed a protective effect. The rs7195409 has a strong association between stage I+II and III+IV in NSCLC, and G allele may play a protective role in the development of NSCLC.

Objective To investigate the preparation of influenza virus vaccine without thimerosal for children dose and its sta-bility .Methods H1N1 ,H3N2 ,B-type influenza virus were inoculated into allantoic fluid to prepare three batches of influenza virus vaccine without thimerosal for children dose and the vaccine stability test was performed .Single radial immunodiffusion(SRID) and enzyme-linked immunosorbent assay(ELISA) were used to detect the concentration of hemagglutinin and egg albumin .Total protein concentration ,appearance ,sterility test ,endotoxin ,free formaldehyde and the pH value of vaccine were also measured .Results The pH value of vaccine was 7 .2 ,with total protein concentration of 182-189 mg/mL .Hemagglutinin concentrations of H1N1 ,H3N2 and B-type influenza virus decreased when they had been placed in 2-8 ℃ for 3 ,6 ,9 ,12 ,18 months or (37 .0 ± 2 .0) ℃ for 7 ,14 days ,however ,they maintained at 6 .0 μg/0 .25 mL or more at last .Conclusion Influenza virus vaccine without thimerosal for chil-dren dose shows improved safty and is accord with the standard of Chinese Pharmacopoeia(2010 edition) .