Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

【Objective】 To identify a case of antibody against highly prevalent antigen through molecular biology technology. 【Methods】 Blood group typing, unexpected antibody identification and cross matching were performed by serological test, and genetic testing of Diego blood group was performed by molecular biology technology. 【Results】 Serological test showed that there was a high prevalence of anti-Di^b in the serum of the patient. Gene sequencing showed that the genotype of the patient was Di(a+b-) . Two cases with Di(a+b-) matched with the patient were screened from 856 blood donors. 【Conclusion】 The combined detection method based on serological test supplemented by molecular biology technology is beneficial to the detection of antibody against highly prevalent antigens, and is of great significance for ensuring the safety of clinical blood transfusion.

Objective:To investigate the changes in the morphology, structure and function of the bladders and their effects on the upper urinary tract dilatation(UUTD) after lumbosacral nerve transecting in rats.Methods:A total of 45 female SD rats were included, randomly divided into 3 groups with 15 rats in each group. Two groups were performed bilateral lumbar 6(L6) and cauda equina nerve shearing to establish neurogenic bladder(NB) model, which were nerve transected for 4 weeks(NB-4W) group and nerve transected for 12 weeks(NB-12W) group. Another group was performed bilateral L6 nerves and cauda equine exposing but not transecting, which was sham-operation (Sham) group. Cystometry and renal ultrasound examination were performed and rats in each group were killed to collect the kidney and bladder tissues in NB-4W group at 4 weeks, in Sham group and NB-12W group at 12 weeks after operation. HE, Masson staining, immunohistochemical staining and western blot were used to detect histological changes, expression of transforming growth factor-β1(TGF-β1) and α-smooth muscle actin(α-SMA).Results:All rats in NB-4W and NB-12W group showed acontractile detrusor. In the NB-4W and NB-12W group, the maximum cystometric capacity [(5.84±0.33) ml and (3.13±0.35) ml], the detrusor leak point pressure [(25.41±0.86) cm H 2O and (27.36±2.04) cm H 2O] (1 cm H 2O = 0.098 kPa), were significantly higher than those in the Sham group [(0.98±0.14) ml, (7.13±0.90) cm H 2O, both P<0.05]. Compliance in NB-4W group [(0.28±0.21) ml/cm H 2O] and NB-12W group [(0.17±0.12) ml/cm H 2O] were significantly lower than that of the Sham group [(0.34±0.26) ml/cm H 2O], and the compliance of NB-12W group was lower than that of NB-4W group significantly (all P<0.05). HE staining of the bladder showed that the inflammatory cell infiltration was obvious in the NB-4W and NB-12W group. Bladder collagen volume fractions in NB-4W group [(30.5±1.5) %] and NB-12W group [(45.2±3.8) %] were both higher than that of Sham group [(20.7±2.2) %, both P<0.05]. The expression of TGF-β1 and α-SMA in the bladder tissue of NB-4W group were higher than those of sham group, and that of NB-12W group were higher than NB-4W group. In NB-4W group and NB-12W group, 3 (20.0 %) and 7 (46.7 %) rats were found hydronephrosis, respectively. Additionally, HE staining showed that the degree of renal tubule injury and the number of inflammatory cell infiltration in the NB-4W and NB-12W group were higher than those in the Sham group. Masson staining showed that the volume fraction of collagen in kidneys of NB-4W and NB-12W group were (13.1±1.4) % and (21.6±1.9) %, respectively, which were significantly higher than that in sham operation group [(4.6±0.7) %, both P<0.05]. Conclusions:Bilateral L6 + cauda equina nerve transecting can induce NB with hydronephrosis in parts of rats. The degree of bladder fibrosis gradually increased with the time of nerve transection, and the incidence and severity of UUTD also increased with the time of nerve transection.

Objective: To investigate the effect of hypochloric acid on Escherichia coli biofilm and the clinical efficacy of hypochloric acid for wounds with Escherichia coli infection. Methods: One strain of Escherichia coli with the strongest bacterial biofilm forming ability among the strains isolated from specimens in 25 patients (16 males and 9 females, aged 32-67 years) from five clinical departments of the 940^th Hospital of the Joint Logistic Support Force was collected for the experimental study from September to December 2019. The Escherichia coli was cultured with hypochloric acid at 162.96, 81.48, 40.74, 20.37, 10.18, 5.09, 2.55, 1.27, 0.64, and 0.32 μg/mL respectively to screen the minimum bactericidal concentration (MBC) of hypochloric acid. The Escherichia coli was cultured with hypochloric acid at the screened MBC for 2, 5, 10, 20, 30, and 60 min respectively to screen the shortest bactericidal time of hypochloric acid. The biofilm formation of Escherichia coli was observed by scanning electron microscopy at 6, 12, 24, 48, 72, and 96 h of incubation, respectively. After 72 h of culture, hypochloric acid at 1, 2, 4, 8, and 16 times of MBC was respectively added to Escherichia coli to screen the minimum biofilm eradicate concentration (MBEC) of hypochloric acid against Escherichia coli. After hypochloric acid at 1, 2, 4, and 8 times of MBEC and sterile saline were respectively added to Escherichia coli for 10 min, the live/dead bacterial staining kit was used to detect the number of live and dead cells, with the rate of dead bacteria calculated (the number of samples was 5). From January to December 2020, 41 patients with infectious wounds meeting the inclusion criteria and admitted to the Department of Burns and Plastic Surgery of the 940^th Hospital of Joint Logistic Support Force of PLA were included into the prospective randomized controlled trial. The patients were divided into hypochloric acid group with 21 patients (13 males and 8 females, aged (46±14) years) and povidone iodine group with 20 patients (14 males and 6 females, aged (45±19) years) according to the random number table. Patients in the 2 groups were respectively dressed with sterile gauze soaked with hypochloric acid of 100 μg/mL and povidone iodine solution of 50 mg/mL with the dressings changed daily. Before the first dressing change and on the 10^th day of dressing change, tissue was taken from the wound and margin of the wound for culturing bacteria by agar culture method and quantifying the number of bacteria. The amount of wound exudate and granulation tissue growth were observed visually and scored before the first dressing change and on the 3^rd, 7^th, and 10^th days of dressing change. Data were statistically analyzed with one-way analysis of variance, Dunnett-t test, independent sample t test, Mann-Whitney U test, Wilcoxon signed-rank test, chi-square test, or Fisher's exact probability test. Results: The MBC of hypochloric acid against Escherichia coli was 10.18 μg/mL, and the shortest bactericidal time of hypochloric acid with MBC against Escherichia coli was 2 min. Escherichia coli was in a completely free state after 6 and 12 h of culture and gradually aggregated and adhered with the extension of culture time, forming a mature biofilm at 72 h of culture. The MBEC of hypochloric acid against Escherichia coli was 20.36 μg/mL. The Escherichia coli mortality rates after incubation with hypochloric acid at 1, 2, 4, and 8 times of MBEC for 10 min were significantly higher than that after incubation with sterile saline (with t values of 6.11, 25.04, 28.90, and 40.74, respectively, P<0.01). The amount of bacteria in the wound tissue of patients in hypochloric acid group on the 10^th day of dressing change was 2.61 (2.20, 3.30)×10⁴ colony forming unit (CFU)/g, significantly less than 4.77 (2.18, 12.48)×10⁴ CFU/g in povidone iodine group (Z=2.06, P<0.05). The amounts of bacteria in the wound tissue of patients in hypochloric acid group and povidone iodine group on the 10^th day of dressing change were significantly less than 2.97 (2.90, 3.04)×10⁶ and 2.97 (1.90, 7.95)×10⁶ CFU/g before the first dressing change (with Z values of 4.02 and 3.92, respectively, P<0.01). The score of wound exudate amount of patients in hypochloric acid group on the 10^th day of dressing change was significantly lower than that in povidone iodine group (Z=2.07, P<0.05). Compared with those before the first dressing change, the scores of wound exudate amount of patients in hypochloric acid group on the 7^th and 10^th days of dressing change were significantly decreased (with Z values of -3.99 and -4.12, respectively, P<0.01), and the scores of wound exudate amount of patients in povidone iodine group on the 7^th and 10^th days of dressing change were significantly decreased (with Z values of -3.54 and -3.93, respectively, P<0.01). The score of wound granulation tissue growth of patients in hypochloric acid group on the 10^th day of dressing change was significantly higher than that in povidone iodine group (Z=2.02, P<0.05). Compared with those before the first dressing change, the scores of wound granulation tissue growth of patients in hypochloric acid group on the 7^th and 10^th days of dressing change were significantly increased (with Z values of -3.13 and -3.67, respectively, P<0.01), and the scores of wound granulation tissue growth of patients in povidone iodine group on the 7^th and 10^th days of dressing change were significantly increased (with Z values of -3.12 and -3.50, respectively, P<0.01). Conclusions: Hypochloric acid can kill Escherichia coli both in free and biofilm status. Hypochloric acid at a low concentration shows a rapid bactericidal effect on mature Escherichia coli biofilm, and the higher the concentration of hypochloric acid, the better the bactericidal effect. The hypochloric acid of 100 μg/mL is effective in reducing the bacterial load on wounds with Escherichia coli infection in patients, as evidenced by a reduction in wound exudate and indirect promotion of granulation tissue growth, which is more effective than povidone iodine, the traditional topical antimicrobial agent.
Adult ; Aged ; Biofilms ; Escherichia coli ; Escherichia coli Infections/drug therapy* ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Surgical Wound Infection ; Treatment Outcome

【Objective】 To investigate the distribution frequency of RBC rare blood group among blood donors in Chongqing, so as to provide basic data for the establishment of regional rare blood group donor database. 【Methods】 A total of 14 805 voluntary blood donors of Chongqing Blood Center from December 2020 to May 2021 were screened for Jk(a-b-) phenotype of Kidd blood group system by urea hemolysis test and confirmed by saline agglutination test. The indirect anti-globulin test was used to screen the Fy(a-) phenotype of Duffy blood group system, s-phenotype of the MNS blood group system and k- phenotype of Kell blood group system in 1 466 O type blood donors. The polyamine test was used to screen the Di(b-) phenotype of Diego blood group system in 856 voluntary blood donors, and confirmed by anti-globulin test. 【Results】 Among the voluntary blood donors in Chongqing, the proportion of Jk(a-b-) phenotype was 0.0203% (3/14 805). The ratio of both Fy(a-b+ ) and S+ s- phenotype among type O blood donors was 0.136 4% (2/1 466), and k- phenotype was not seen. The proportion of Di(a+ b-) phenotype among 856 blood donors was 0.233 6% (2/826). 【Conclusion】 The distribution frequency of rare blood group antigens in the above five blood group systems in Chongqing voluntary blood donors presents regional characteristics.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective To analyze stress distributions of alveolar bone around implants with different types of implant thread shapes and thread depths during dental immediate implantation, so as to provide references for the design and selection of implants. Methods The simplified model of mandible bone block, implants and mandibular molar were established by Geomagic Studio, SolidWorks and ANSYS Workbench, and vertical and oblique loads were applied on this model, respectively. Stress distributions on implants with different thread shape and thread depth as well as alveolar bone around implants were calculated. Results Under vertical loading, the peak stresses of implants, cortical bone and cancellous bone were in the range of 120.51-129.63 MPa, 9.94-13.25 MPa and 3.92-8.01 MPa, respectively. And the stress of cortical bone around V-shaped, rectangular, buttress or reverse buttress implant remained stable in the range of 0.40-0.45 mm thread depth. Under oblique loading, the peak stresses of implants, cortical bone and cancellous bone were in the range of 220.23-286.51 MPa, 33.39-45.08 MPa, 4.96-12.5 MPa, respectively. Among the models, V-shaped, buttress, reverse buttress implant with 0.45 mm thread depth showed the minimum stress. Conclusions The V-shaped, buttress or reverse buttress implant with a thread depth of 0.45 mm, or the rectangular implant with a thread depth of 0.40 mm had better biomechanical properties.