AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

Objectives miR-207 has been identified as being expressed in natural killer (NK) cell exosomes that play a role in disease progression; however, to date, there are no studies specifically linking miR-207 to tuberculosis (TB). Methods Bioinformatics methods employed for prediction, followed by a dual luciferase reporter assay to determine whether lysosome-associated membrane protein 2 (LAMP2) is targeted by miR-207. The experiments were divided into four groups using the liposome transfection method (OP-LAMP2 group: co-transfected with miR-207 mimics and LAMP2 overexpression plasmid; EP group: co-transfected with mimics NC and null-loaded plasmid; siLAMP2 group: transfected with siLAMP2; and siLAMP2-NC group: transfected with siLAMP2-NC). TB infection was modeled using H37Ra-infected Ana-1 cells. The impact of LAMP2 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria were assessed by tuberculosis colony-forming unit counting. Flow cytometry was used to assess the total apoptosis rate. Real-time fluorescent quantitative PCR was conducted to determine the relative expression of LAMP2, apoptosis genes, pyroptosis genes, and autophagy genes. Western blot analysis was performed to measure the relative expression of LAMP2 proteins, apoptosis proteins, pyroptosis proteins, and autophagy proteins. Results Dual luciferase reporter assay test showed that there was a targeting relationship between LAMP2 and miR-207. The transfection model was successfully constructed under real-time fluorescent quantitative PCR and Western blot statistical analysis, and microscopic observation. The infection model was successfully established under microscopic observation. Colony forming unit counting revealed that the number of colonies in the OP-LAMP2 group was lower than that in the EP group, while the number of colonies in the siLAMP2 group was higher than that in the siLAMP2-NC group. Flow cytometry assay revealed that the total apoptosis in OP-LAMP2 group was lower than that in EP group, and the total apoptosis in siLAMP2 group was higher than that in siLAMP2-NC group. Real-time fluorescence quantitative PCR and Western blot analysis revealed that the relative expression of apoptosis and pyroptosis-related proteins and genes in the control group was lower in the OP-LAMP2 group compared to the EP group, and higher in the siLAMP2 group compared to the siLAMP2-NC group. Real-time fluorescence quantitative PCR detected that the relative expression of autophagy positively regulated genes Microtubule-associated protein 1 light chain 3(LC3)and Beclin1 in the OP-LAMP2 group was higher in the OP-LAMP2 group compared to the EP group, and lower in the siLAMP2 group compared to the siLAMP2-NC group, while the relative expression of negatively regulated autophagy genes followed the opposite trend to that of autophagy positively regulated genes. The relative expression of autophagy-related proteins was consistent with the trend of autophagy genes. Conclusions miR-207 enhances macrophage apoptosis, cellular pyroptosis and inhibits autophagy, promoting survival of Mycobacterium tuberculosis by targeting the autophagy-related protein LAMP2, thus offering a novel therapeutic direction for tuberculosis.
Lysosomal-Associated Membrane Protein 2/metabolism* ; MicroRNAs/metabolism* ; Mycobacterium tuberculosis/physiology* ; Autophagy/genetics* ; Humans ; Macrophages/metabolism* ; Apoptosis/genetics* ; Tuberculosis/metabolism* ; Cell Line ; Pyroptosis/genetics*

Pulmonary diseases, as a prevalent category of respiratory system disorders, have become a significant global public health concern. The increasing incidence of these diseases, caused by environmental pollution and occupational hazards, poses a substantial threat to human health and the overall quality of life. Mesenchymal stem cells (MSCs) are known for their remarkable immunomodulatory, anti-bacterial, and anti-apoptotic capabilities. Exosomes derived from MSCs, carrying a diverse array of proteins, lipids, nucleic acids, and other bio-active molecules, have demonstrated considerable therapeutic potential in treating pulmonary diseases, and have come to the forefront of medical research. This review summarized the therapeutic role of exosomes derived from various sources of mesenchymal stem cells in the context of pulmonary diseases, aiming to provide a robust foundation for their clinical application in diagnosis and treatment.
Exosomes/transplantation* ; Humans ; Mesenchymal Stem Cells/metabolism* ; Lung Diseases/therapy* ; Animals

Objective To investigate the antioxidant activity of bamboo stem extracts and the therapeutic effect of bamboo stem water extract on oxidative inflammation induced by tert butyl hydroperoxide (t-BHP) in human colon adenocarcinoma cells (Caco-2). Methods In this study, ABTS, DPPH, and FRAP assays were used to determine the extracellular antioxidant activity of petroleum ether extract, ethyl acetate extract, n-butanol extract, 95% ethanol extract, and distilled water extract from bamboo stems. The human intestinal Caco-2 cell line was used as the model cell, and t-BHP was selected as the oxidative stress modeling agent. The CCK-8 assay was used to detect cell viability and the optimal oxidative damage concentration of t-BHP. The content of MDA, 8-OHdG, TNF-α and IL-1β were detected to assess antioxidant stress effect. Results The five extracts of bamboo all had certain antioxidant activity, among which the water extract of bamboo stem had the best comprehensive antioxidant activity with high cell viability in Caco-2 cells. The optimal modeling concentration of t-BHP was 200 μMol/L. The water extract of bamboo stem significantly reduced the content of oxidative stress related biomarkers and inflammatory factors in Caco-2 cells induced by t-BHP. Conclusion The stem extracts of bamboo in Xianning City have strong in vitro antioxidant activity. Among them, the water extract of bamboo stem has a protective effect on t-BHP induced oxidative damage in Caco-2 cells, suggesting that the water extract possesses a potential to be developed as new antioxidant products for clinical prevention and treatment of oxidative damage related diseases.

Objective:By conducting genetic testing of hereditary hearing loss in pregnant women within 17 weeks of gestation in Dali areas, the importance of genetic testing and genetic counseling during pregnancy was emphasized. Methods:Twenty-one mutation sites of 4 hearing loss genes, including GJB2, GJB3, SLC26A4 and mtDNA, were detected by PCR amplification technology. The positive ratio, mutation ratio and ethnic distribution of positive samples were statistically described. Results:The positive ratios of GJB2 and SLC26A4 genes were 1.24% and 1.43%, respectively, with mutation rates of 40.62% and 46.88% in the positive samples, respectively. The positive ratio of GJB3gene was 0.19%, and mtDNA mutation genes accounted for 0.14%, and all of them were mtDNA（Heterozygous）. There was only one case of GJB2/SLC26A4 double positive multi-gene mutation, with a positive ratio of 0.05%. The frequency of GJB2 c. 235delC site was the highest, accounting for 65.38% of GJB2 mutation genes and 26.56% of mutation gene samples. Conclusion:GJB2 and SLC26A4 are the most common genes of hearing loss, and GJB2 c. 235delC site is the most common mutation site. Identifying the hearing loss mutation site is of great importance to prevent the birth of hereditary hearing loss children, and genetic diagnosis, genetic counseling, and appropriate intervention are crucial to alleviate congenital problems.
Humans ; Female ; Pregnancy ; Sulfate Transporters/genetics* ; Connexin 26 ; Genetic Testing/methods* ; Connexins/genetics* ; Mutation ; Hearing Loss/diagnosis* ; DNA, Mitochondrial/genetics* ; Adult ; Membrane Transport Proteins/genetics* ; Genetic Counseling

Objective:This study aimed to explore the clinical application value of the indication selection criteria for bi-pedicled deep inferior epigastric arterial perforator flap(DIEP) versus pedicled transverse rectus abdominis myocutaneous flap(TRAM) combined with DIEP for breast reconstruction.Methods:The clinical data of patients who underwent delayed bi-pedicled abdominal flap breast reconstruction after breast cancer surgery in the Department of Oncoplastic and Reconstructive Breast Surgery and Department of Mammoplasty, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2008 to April 2022 were retrospectively analyzed. According to the indications for each procedure, corresponding blood supply source was selected. Subsequently, patients were divided into two groups: bi-pedicled DIEP group and pedicled TRAM+ DIEP combined flap group according to the type of blood supply source. The indication selection criteria for pedicled TRAM+ DIEP combined flap was as follows: lack of two groups of recipient vessels; no dominant perforator in the abdominal donor site; previous abdominal liposuction or abdominal surgery with large dissection range and severe scar; patients were unable to tolerate prolonged surgery or had high risk factors for microsurgery. A comparison of breast reconstruction surgery characteristics was conducted between the two groups. Independent-samples t-test, rank sum test, Chi-square test and Fisher’s exact probability method were used to analyze various data including age, body mass index (BMI), time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy, history of chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length, preoperative and postoperative conditions, as well as postoperative complications. Statistical significance was defined at P<0.05. Results:A total of 54 patients who underwent unilateral breast reconstruction were included in this study with a mean age of 42.2 years (range, 30-59 years). The pedicled TRAM+ DIEP combined flap group consisted of 21 patients with a mean age of 42.7 years (range, 33-56 years) while the bi-pedicled DIEP group comprised 33 patients with a mean age of 41.8 years (30-59 years). Out of the 54 patients, a total of 38 were eventually followed up, comprising 14 in the pedicled TRAM+ DIEP combined flap group and 24 in the bi-pedicled DIEP group. The average follow-up duration for the pedicled TRAM+ DIEP combined flap group was 42 months (range, 6-69 months). Abdominal bulge occurred in one patient, while another patient experienced abdominal hernia. The average follow-up duration for the bi-pedicled DIEP group was 47 months (6-179 months), with no reported cases of abdominal bulge or abdominal hernia. There were no statistically significant differences observed between the two groups regarding age, BMI, time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy or chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length or time to ambulation (all P>0.05). However, there was a statistically significant difference noted in duration of surgery between the pedicled TRAM+ DIEP combined flap group and bi-pedicled DIEP group [6.0(5.5-6.5) hours vs. 8.5(8.0-8.8) hours] ( P<0.01). Comparison analysis revealed no statistically significant differences in terms of partial flap necrosis (0/21 vs. 1/33), abdominal incision dehiscence (2/21 vs. 2/33), abdominal bulge (1/14 vs. 0 /24) or abdominal hernia (1/1 vs. 0/24) between the two groups ( P>0.05). Conclusion:For patients who needed bilateral vascular pedicled lower abdominal wall for breast reconstruction, utilizing a pedicled TRAM+ DIEP combined flap did not increase surgical complication risks. The proposed indication selection criteria for using a pedicled TRAM + DIEP combined flap outlined in this study could serve as guidance when choosing methods for bi-pedicled abdominal flap breast reconstruction.

Objective:To preliminarily evaluate the outcome of fat grafting breast augmentation using filtration-adsorption technique.Methods:Patients undergoing autologous fat grafting (AFG) breast augmentation between July of 2020 and March of 2021 were retrospectively reviewed at Breast Plastic and Reconstructive Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College. Patients with complete data who followed up for at least 3 months were included. The lipoaspirates were processed with filtration-adsorption technique and AFG breast augmentation were performed (filtration-adsorption group), and then sedimentation method which was commonly used clinically was set as the control group (sedimentation group). During the follow-up, breast palpation and ultrasound inspection were performed, and the breast volumetric change between preoperative and postoperative was quantitatively compared by three-dimensional scanning technique. Demographic and surgical details were statistically analyzed. Postoperative breast volume retention rates and complication rates were calculated and compared. Normally distributed continuous variables were described in the form Mean±SD, and t-test was used for analysis. Non-normal distributed continuous variables were described in the form of M ( Q1, Q3) and analyzed by Mann-Whitney U test. The classified data were analyzed by Fisher exact probability test. Results:A total of 20 female patients (40 breasts) were included, including 10 patients (20 breasts) aged (31.4±2.5) years in sedimentation group and 10 patients (20 breasts) aged (28.5±4.4) years in the filtration-adsorption group. The fat injection volume in the filtration-adsorption group was 151.1 (125.0, 175.0) ml, and 165.0 (151.3, 196.3) ml in sedimentation group, respectively, with no statistical significance ( P>0.05). The breast volume retention rate in filtration-adsorption group was (62.93±14.06)%, which was significantly higher than that in sedimentation group (24.97±11.02)% ( P<0.01). During postoperative breast ultrasound examination, 2 breasts in sedimentation group had palpable nodules (< 6 mm), 1 breast had ultrasonically detectable but clinically inaccessible nodules (< 4 mm), while only 1 breast in the filtration-adsorption group had an oil cyst (4 mm×2 mm). The complication rate in the filtration-adsorption group was lower than that in sedimentation group[5.0%(1/20) vs. 15.0%(3/20), P>0.05]. Conclusion:The filtration-adsorption technique is an ideal method for fat grafting, which would multiply the volume retention rate in large-volume AFG breast augmentation and reduce the rate of complications.

Objective:This study aimed to explore the clinical application value of the indication selection criteria for bi-pedicled deep inferior epigastric arterial perforator flap(DIEP) versus pedicled transverse rectus abdominis myocutaneous flap(TRAM) combined with DIEP for breast reconstruction.Methods:The clinical data of patients who underwent delayed bi-pedicled abdominal flap breast reconstruction after breast cancer surgery in the Department of Oncoplastic and Reconstructive Breast Surgery and Department of Mammoplasty, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2008 to April 2022 were retrospectively analyzed. According to the indications for each procedure, corresponding blood supply source was selected. Subsequently, patients were divided into two groups: bi-pedicled DIEP group and pedicled TRAM+ DIEP combined flap group according to the type of blood supply source. The indication selection criteria for pedicled TRAM+ DIEP combined flap was as follows: lack of two groups of recipient vessels; no dominant perforator in the abdominal donor site; previous abdominal liposuction or abdominal surgery with large dissection range and severe scar; patients were unable to tolerate prolonged surgery or had high risk factors for microsurgery. A comparison of breast reconstruction surgery characteristics was conducted between the two groups. Independent-samples t-test, rank sum test, Chi-square test and Fisher’s exact probability method were used to analyze various data including age, body mass index (BMI), time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy, history of chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length, preoperative and postoperative conditions, as well as postoperative complications. Statistical significance was defined at P<0.05. Results:A total of 54 patients who underwent unilateral breast reconstruction were included in this study with a mean age of 42.2 years (range, 30-59 years). The pedicled TRAM+ DIEP combined flap group consisted of 21 patients with a mean age of 42.7 years (range, 33-56 years) while the bi-pedicled DIEP group comprised 33 patients with a mean age of 41.8 years (30-59 years). Out of the 54 patients, a total of 38 were eventually followed up, comprising 14 in the pedicled TRAM+ DIEP combined flap group and 24 in the bi-pedicled DIEP group. The average follow-up duration for the pedicled TRAM+ DIEP combined flap group was 42 months (range, 6-69 months). Abdominal bulge occurred in one patient, while another patient experienced abdominal hernia. The average follow-up duration for the bi-pedicled DIEP group was 47 months (6-179 months), with no reported cases of abdominal bulge or abdominal hernia. There were no statistically significant differences observed between the two groups regarding age, BMI, time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy or chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length or time to ambulation (all P>0.05). However, there was a statistically significant difference noted in duration of surgery between the pedicled TRAM+ DIEP combined flap group and bi-pedicled DIEP group [6.0(5.5-6.5) hours vs. 8.5(8.0-8.8) hours] ( P<0.01). Comparison analysis revealed no statistically significant differences in terms of partial flap necrosis (0/21 vs. 1/33), abdominal incision dehiscence (2/21 vs. 2/33), abdominal bulge (1/14 vs. 0 /24) or abdominal hernia (1/1 vs. 0/24) between the two groups ( P>0.05). Conclusion:For patients who needed bilateral vascular pedicled lower abdominal wall for breast reconstruction, utilizing a pedicled TRAM+ DIEP combined flap did not increase surgical complication risks. The proposed indication selection criteria for using a pedicled TRAM + DIEP combined flap outlined in this study could serve as guidance when choosing methods for bi-pedicled abdominal flap breast reconstruction.

Objective:To preliminarily evaluate the outcome of fat grafting breast augmentation using filtration-adsorption technique.Methods:Patients undergoing autologous fat grafting (AFG) breast augmentation between July of 2020 and March of 2021 were retrospectively reviewed at Breast Plastic and Reconstructive Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College. Patients with complete data who followed up for at least 3 months were included. The lipoaspirates were processed with filtration-adsorption technique and AFG breast augmentation were performed (filtration-adsorption group), and then sedimentation method which was commonly used clinically was set as the control group (sedimentation group). During the follow-up, breast palpation and ultrasound inspection were performed, and the breast volumetric change between preoperative and postoperative was quantitatively compared by three-dimensional scanning technique. Demographic and surgical details were statistically analyzed. Postoperative breast volume retention rates and complication rates were calculated and compared. Normally distributed continuous variables were described in the form Mean±SD, and t-test was used for analysis. Non-normal distributed continuous variables were described in the form of M ( Q1, Q3) and analyzed by Mann-Whitney U test. The classified data were analyzed by Fisher exact probability test. Results:A total of 20 female patients (40 breasts) were included, including 10 patients (20 breasts) aged (31.4±2.5) years in sedimentation group and 10 patients (20 breasts) aged (28.5±4.4) years in the filtration-adsorption group. The fat injection volume in the filtration-adsorption group was 151.1 (125.0, 175.0) ml, and 165.0 (151.3, 196.3) ml in sedimentation group, respectively, with no statistical significance ( P>0.05). The breast volume retention rate in filtration-adsorption group was (62.93±14.06)%, which was significantly higher than that in sedimentation group (24.97±11.02)% ( P<0.01). During postoperative breast ultrasound examination, 2 breasts in sedimentation group had palpable nodules (< 6 mm), 1 breast had ultrasonically detectable but clinically inaccessible nodules (< 4 mm), while only 1 breast in the filtration-adsorption group had an oil cyst (4 mm×2 mm). The complication rate in the filtration-adsorption group was lower than that in sedimentation group[5.0%(1/20) vs. 15.0%(3/20), P>0.05]. Conclusion:The filtration-adsorption technique is an ideal method for fat grafting, which would multiply the volume retention rate in large-volume AFG breast augmentation and reduce the rate of complications.