Objective To understand the distribution and antimicrobial resistance of bacteria isolated from bronchoalveolar lavage fluid (BALF) in Sichuan Province over a five-year period,and provide a reference for antimicrobial therapy.Methods Bacteria were identified and subjected to antimicrobial susceptibility testing (including disk diffusion and automated systems) according to the protocol of the China Antimicrobial Resistance Surveillance System (CARSS).The results were interpreted according to the breakpoints recommended in CLSI M100 (2022 Edition).Results A total of 26575 strains of bacteria were included in this study,of which 83.2％ were Gram-negative bacteria and 16.8％ were Gram-positive bacteria.The top bacteria species were Klebsiella pneumoniae (19.8％),Pseudomonas aeruginosa (18.5％),Acinetobacter baumannii (12.3％),Staphylococcus aureus (7.7％),and Stenotrophomonas maltophilia (6.7％).None of S.aureus strains were resistant to vancomycin,teicoplanin,or linezolid.The prevalence of methicillin-resistant S.aureus (MRSA) was 32.5％.The resistance rate of Streptococcus pneumoniae to penicillin was 1.5％,and the resistance rate to erythromycin and clindamycin was>90％.The prevalence of β-lactamase-producing strains was 63.3％ in Haemophilus influenzae,while>90％ of H.influenzae strains were susceptible to levofloxacin.Less than 5％ of E.coli strains were resistant to carbapenems,and less than 15％ of the strains were resistant to cefoperazone-sulbactam and piperacillin-tazobactam.The resistance rate of K.pneumoniae to carbapenems increased year by year,from 11.1％ in 2019 to 19.1％ in 2023.The resistance rate of A.baumannii to most antimicrobial agents was>50％.The resistance rate of P.aeruginosa to carbapenems was about 15％.Conclusions The main bacteria isolated from BALF specimens in Sichuan Province are Gram-negative bacteria.The antimicrobial resistance of the bacterial isolates is on the rise.For the patients suspected of lower respiratory tract infection,clinical practitioners should collect more high-quality BALF specimens for microbiology testing.Surveillance of the distribution and antimicrobial susceptibility of the pathogens in respiratory tract infections can inform better empirical treatment.

Objective This study aims to investigate the influence of glucose regulated protein(GRP)78 on osteo-blast differentiation in periodontal ligament fibroblasts(PDLFs)under cyclic mechanical stretch and determine the under-lying mechanism.Methods FlexCell 5000 cell mechani-cal device was applied to simulate the stress environment of orthodontic teeth.GRP78High and GRP78Low sub-population were obtained by flow sorting.Gene transfec-tion was performed to knockdown GRP78 and c-Src ex-pression and overexpress c-Src.Western blot analysis was used to detect the protein expression of Runt-related gene 2(RUNX2),Osterix,osteocalcin(OCN),and osteopontin(OPN).Immunoprecipitation assay was used to determine the interaction of GRP78 with c-Src.The formation of cellular mineralized nodules was determined by alizarin red staining.Results GRP78 was heterogeneously expressed in PDLFs,and GRP78High and GRP78Low subpopulations were obtained by flow sorting.The osteogenic differentiation ability and phosphorylation level of c-Src kinase in the GRP78High subpopulation were significantly increased com-pared with those in GRP78Low subpopulation after cyclic mechanical stretch(P<0.05).GRP78 interacted with c-Src in PDLFs.The overexpression c-Src group showed significantly increased osteogenic differentiation ability than the vector group(P<0.05),and the sic-Src group showed significantly decreased osteogenic differentiation ability(P<0.05)after cy-clic mechanical stretch.Conclusion GRP78 upregulates c-Src expression by interacting with c-Src kinase and pro-motes osteogenic differentiation under cyclic mechanical stretch in PDLFs.

Objective To understand the distribution and antimicrobial resistance of bacteria isolated from bronchoalveolar lavage fluid (BALF) in Sichuan Province over a five-year period,and provide a reference for antimicrobial therapy.Methods Bacteria were identified and subjected to antimicrobial susceptibility testing (including disk diffusion and automated systems) according to the protocol of the China Antimicrobial Resistance Surveillance System (CARSS).The results were interpreted according to the breakpoints recommended in CLSI M100 (2022 Edition).Results A total of 26575 strains of bacteria were included in this study,of which 83.2％ were Gram-negative bacteria and 16.8％ were Gram-positive bacteria.The top bacteria species were Klebsiella pneumoniae (19.8％),Pseudomonas aeruginosa (18.5％),Acinetobacter baumannii (12.3％),Staphylococcus aureus (7.7％),and Stenotrophomonas maltophilia (6.7％).None of S.aureus strains were resistant to vancomycin,teicoplanin,or linezolid.The prevalence of methicillin-resistant S.aureus (MRSA) was 32.5％.The resistance rate of Streptococcus pneumoniae to penicillin was 1.5％,and the resistance rate to erythromycin and clindamycin was>90％.The prevalence of β-lactamase-producing strains was 63.3％ in Haemophilus influenzae,while>90％ of H.influenzae strains were susceptible to levofloxacin.Less than 5％ of E.coli strains were resistant to carbapenems,and less than 15％ of the strains were resistant to cefoperazone-sulbactam and piperacillin-tazobactam.The resistance rate of K.pneumoniae to carbapenems increased year by year,from 11.1％ in 2019 to 19.1％ in 2023.The resistance rate of A.baumannii to most antimicrobial agents was>50％.The resistance rate of P.aeruginosa to carbapenems was about 15％.Conclusions The main bacteria isolated from BALF specimens in Sichuan Province are Gram-negative bacteria.The antimicrobial resistance of the bacterial isolates is on the rise.For the patients suspected of lower respiratory tract infection,clinical practitioners should collect more high-quality BALF specimens for microbiology testing.Surveillance of the distribution and antimicrobial susceptibility of the pathogens in respiratory tract infections can inform better empirical treatment.

Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
China ; Cystic Fibrosis/genetics* ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics* ; Humans ; Mutation ; Poly T ; RNA, Messenger/genetics*

Objective:To explore the selection and effects of flap/myocutaneous flap repair methods for the defect after perineum tumor resection.Methods:From January 2011 to February 2017, 31 patients with vulvar tumor who were admitted to Hunan Cancer Hospital underwent repair of wound after tumor resection with various flaps/myocutaneous flaps. The patients were composed of 5 males and 26 females, aged 39-76 years, with 27 vulvar cancer and 4 Paget′s disease in primary diseases. The size of defects after vulvar tumor radical resection ranged from 8.0 cm×4.5 cm to 27.5 cm×24.0 cm. According to the theory of perforasome, the defects were repaired by the external pudendal artery perforator flap, deep inferior epigastric artery perforator flap, rectus abdominis myocutaneous flap, anterolateral thigh flap, internal pudendal artery perforator flap, gracilis myocutaneous flap, and profunda artery perforator flap based on the specific size and location of perineum and groin where the defect was located. According to the blood supply zone of flap, totally 17 local translocation flaps, 18 axial flaps/myocutaneous flaps, and 7 V-Y advancement flaps were resected, with an area of 7.0 cm×4.0 cm to 21.0 cm×13.0 cm. All the flaps/myocutaneous flaps were transferred in pedicled fashion, and the donor sites were closed without tension. The number of flaps/myocutaneous flaps, wound closure, flaps/myocutaneous flaps survival, and follow-up were observed and recorded.Results:Altogether 42 flaps/myocutaneous flaps were harvested in 31 patients. Two flaps/myocutaneous flaps were used in 11 cases for large circular defect repair. All the defects achieved tension-free primary closure. The blood supply of 32 flaps/myocutaneous flaps was good, while insufficient blood supply was noted in the other 10 flaps/myocutaneous flaps. Seventeen flaps/myocutaneous flaps survived smoothly. Wound dehiscence occurred in 5 flaps/myocutaneous flaps 8 to 14 days postoperatively, which was healed with dressing change. Temporary congestion was noted in 7 flaps/myocutaneous flaps 2 to 5 days postoperatively, which recovered without special treatment. Three flaps/myocutaneous flaps had infection 7 to 15 days postoperatively, two of which recovered after dressing change, while the other one had partial necrosis and received debridement and direct closure. Two flaps/myocutaneous flaps were totally necrotic 8 to 15 days postoperatively, which were repaired with pedicled rectus abdominis myocutaneous flap after debridement. Seven flaps/myocutaneous flaps had partial necrosis 7 to 20 days postoperatively and were healed after dressing change. Twenty-four patients were followed up for 9-38 months. The color of flaps/myocutaneous flaps was similar to that of the surrounding skin, the shape of vulva was natural, the movement of hip joint was not limited, the function of micturition and defecation was not affected, and tumor recurrence was noted in 3 patients.Conclusions:For the complicated large defect after perineum tumor resection, the flexible application of different forms of flaps/myocutaneous flaps to repair according to different areas regains the appearance and function. However, there are many complications, so it is necessary to further strengthen the postoperative care.

Objective:To explore the extending design and anatomical basis of pedicled gracilis musculocutaneous flaps in perineal reconstruction.Methods:On six female cadavers (age ranged from 27 to 52 years old, 41.2 on average), bilateral gracilis flaps were designed, raised and transferred in the standard surgical manner and tunneled under the adductor longus muscle. Measured the distance from the tip of 12 flaps to the bilateral anterior superior iliac spine (ASIS). The myocutaneous flaps were then transferred and the distances from the tip of flaps to the ASIS were measured again. The paired sample t test (double tail) was used to compare the data measured by the two transfer method.Results:Cadaveric dissection revealed that in 6 corpses, bilateral profunda artery muscular branches all courses between the longus adductor and mangus adductor muscles, the pedicle length of 12 gracilis musculocutaneous flaps was(7.84±0.71) cm. In the standard surgical manner group, after transferring, the distance from the tip of flaps to the ipsilateral ASIS was (9.23±0.95) cm, ranged from 6.9 cm to 10.5 cm, the distance from the tip of flaps to the contralateral ASIS was(11.45±2.27) cm, ranged from 7.5 cm to 14.8 cm. In the tunnelled under the adductor longus muscle extension design group, after transferring, the distance from the tip of flaps to the ipsilateral ASIS was (4.52±1.18) cm, ranged from 2.7 cm to 6.5 cm, the distance from the tip of flaps to the contralateral ASIS was (8.92 ±1.82) cm, ranged from 4.8 cm to 11.7 cm. Compared with the standard surgical manner group, tunneling the flap under the adductor longus muscle to the ipsilateral side increased the reach by 3.5 cm to 7.3 cm, (4.71±1.14) cm, to contralateral side increased the reach by 0.9 cm to 4.1 cm, (2.53±0.89) cm. The distance to the ipsilateral anterior superior iliac spine measured by the two method , t=13.194, P<0.001, and the distance to the contralateral anterior superior iliac spine, t=9.057, P<0.001. The difference was statistically significant. The tunneled under the adductor longus muscle extension design group can increase the reach of reconstructive scope by 3.62 cm on average. The additional increasing length of tunneled under the adductor longus muscle method is 46.2%(3.62/7.84) of the average vascular pedicle length. Conclusions:Cadaveric dissection result have shown that tunneling of the gracilis musculocutaneous flap in a novel extension design way can enlengthen the pedicle length, increase its reach across the perineum.

Objective:To explore the extending design and anatomical basis of pedicled gracilis musculocutaneous flaps in perineal reconstruction.Methods:On six female cadavers (age ranged from 27 to 52 years old, 41.2 on average), bilateral gracilis flaps were designed, raised and transferred in the standard surgical manner and tunneled under the adductor longus muscle. Measured the distance from the tip of 12 flaps to the bilateral anterior superior iliac spine (ASIS). The myocutaneous flaps were then transferred and the distances from the tip of flaps to the ASIS were measured again. The paired sample t test (double tail) was used to compare the data measured by the two transfer method.Results:Cadaveric dissection revealed that in 6 corpses, bilateral profunda artery muscular branches all courses between the longus adductor and mangus adductor muscles, the pedicle length of 12 gracilis musculocutaneous flaps was(7.84±0.71) cm. In the standard surgical manner group, after transferring, the distance from the tip of flaps to the ipsilateral ASIS was (9.23±0.95) cm, ranged from 6.9 cm to 10.5 cm, the distance from the tip of flaps to the contralateral ASIS was(11.45±2.27) cm, ranged from 7.5 cm to 14.8 cm. In the tunnelled under the adductor longus muscle extension design group, after transferring, the distance from the tip of flaps to the ipsilateral ASIS was (4.52±1.18) cm, ranged from 2.7 cm to 6.5 cm, the distance from the tip of flaps to the contralateral ASIS was (8.92 ±1.82) cm, ranged from 4.8 cm to 11.7 cm. Compared with the standard surgical manner group, tunneling the flap under the adductor longus muscle to the ipsilateral side increased the reach by 3.5 cm to 7.3 cm, (4.71±1.14) cm, to contralateral side increased the reach by 0.9 cm to 4.1 cm, (2.53±0.89) cm. The distance to the ipsilateral anterior superior iliac spine measured by the two method , t=13.194, P<0.001, and the distance to the contralateral anterior superior iliac spine, t=9.057, P<0.001. The difference was statistically significant. The tunneled under the adductor longus muscle extension design group can increase the reach of reconstructive scope by 3.62 cm on average. The additional increasing length of tunneled under the adductor longus muscle method is 46.2%(3.62/7.84) of the average vascular pedicle length. Conclusions:Cadaveric dissection result have shown that tunneling of the gracilis musculocutaneous flap in a novel extension design way can enlengthen the pedicle length, increase its reach across the perineum.

Objective: To understand the relationship between chloroquine resistance and the virulence of Plasmodium berghei. Met hods: Dynamic changes of histopathologic features of livers, spleens, brains, hearts, lungs and kidneys of mice infected with the chloroquine-sensitive (N) and the chloroquine-resistant (RC) strains of P. berghei were compared. Results: In mice infected with the N strain, deposition of heavy hemoz oin in livers and spleens, congestive edema in lungs, and congestion and embolis m in the brain capillaries were observed. The histopathologic features revealed ac ute inflammatory reaction. In mice infected with the RC strain, histopathologic variations of livers and spleens were associated with changes of parasitemia. In terstitial pneumonia was displayed in lungs. There were chronic histopathologic changes of the organs in the mice infected with RC strain. Conclusion: The mice infected by the N strain with potent virulence die due to adher ence of the erythrocytes to microvascular endothelia and embolism of the microva scula, especially in their brain. Immune responses of the mice infected by the R C strain with poor virulence may be a delayed-type hypersensitive inflammation a ssociated with CD4+Th1 at an early stage of the infection, but may become anti body-dependent immune response assisted with CD4+Th2, which play a key role in elimination of the malaria parasites at later stage of the infection.

AIM:To investigate the depressant effect and mechanism of atorvastatin on the chronic rejection of aortic allograft in rats.METHODS:The models of abdominal aorta transplantation were made with micro-surgery in rats.The recipients were divided into three groups:allograft control group,atorvastatin-treated group and isograft control group.Vascular intimal thickness in all of the groups was observed by histological examination.The expression of proliferation cell nuclear antigenl(PCNA)and ?-smooth muscle actin(?-SMA)were determined by immunohistochemistry.The content of nitric oxide was measured by nitrate reductase chromatometry.RESULTS:The vascular intimal thickness in atorvastatin-treated group(11.60%?2.40%)was lower than that in allograft control group(34.60%?6.40%,P

Objective To study the depressant effect of atorvastatin on arteriosclerosis of aortic allograft in rats.Methods The models of abdominal aorta transplantation were established in rats with the use of(micro-surgery).The recipients were divided into three groups:allograft control group,allograft experimental group and isograft control group.After 60 days of transplant,vascular intimal thickness(VIT) in all of the groups was observed by histological examination.The expression of PCNA and ?-SMA was determined by(immunohistochemistry).Results The degree of VIT in rats of the allograft experimental group was lower than that in the allograft control group;the VIT area ratio in the allograft control group,allograft experimental group and isograft control group was(12.40?2.65)%,(5.20?6.35)%,and(1.2?1.10)%,respectively,A statistical difference between these groups was observed(P