Objective To compare on the influence of invigorating spleen and restoring virility to the SS level and SSR1 gene expression in brain. Method The spleen insufficiency model of rat was established by adopting bitter decline diarrhea, wild eating and overworking. SS level and SSR1 gene expression change of hypothalamus ventral kernel, hippocampi CA1 area and forehead corte were detected by immunohistochemistry and in situ hybridization techniques. Result In the model group, the immunoreaction masculine substance of SS and SSR1 mRNA, hypothalamus ventral kernel, hippocampi CA1 area and forehead cortex decreased obviously. The immunoreaction masculine substance of SS and SSR1 mRNA on above mentioned parts in the group of invigorating spleen and restoring virility increased distinctly. Conclusion In the model group, the SS level and SSR1 gene expression in brain to promoting study memory ability changed. Invigorating spleen and restoring virility may affect study memory ability by regulating the SS level and SSR1 gene expression.

Objective To improve the diagnosis of tuberculosis ( TB) by analyzing Mycobacterium infection in fine-needle aspiration biopsy specimens from children with tuberculous lymphadenitis .Methods Fine-needle aspiration biopsy was performed on 269 children with tuberculous lymphadenitis diagnosed by Shanghai Public Health Clinical Center from January 2011 to September 2013 .The needle aspiration biopsy specimens were processed for acid-fast bacillus (AFB) smear test, mycobacterial culture and Mycobacterium identification ( p-nitrobenzoic acid inhibition test ) .Results Cytological diagnosis of tuberculous lymphade-nitis was made for 269 patients.The positive results by AFB smear test were detected in 63.19% of 269 specimens (n=170) and 40.15%(n=108) specimens were positive in mycobacterial culture .The differ-ence between the two tests were significant (P<0.01).The positive rate of Mycobacterium detected by using BACTEC MGIT 960 automated system and L?wenstein-Jensen culture method were 38 .66% ( n=104 ) and 28.99%(n=78), respectively, showing the significant difference between two tests (P<0.05).AFB smear test in combination with mycobacterial culture could precisely diagnose 70.63% of tuberculous lym-phadenitis in children.Of the 108 clinical isolates, 105 strains (97.2%) were Mycobacterium tuberculosis complex and the rest were non-tuberculous Mycobacterium strains (2.8%).Conclusion The positive rate by AFB smear test was significantly increased in fine needle aspiration biopsy specimens after a series of treatments including sample digestion , centrifugation and precipitation , but the positive rate of mycobacterial culture was reduced .Diagnostic accuracy could be significantly improved by using BACTEC MGIT 960 sys-tem.Mycobacterium tuberculosis complex was the predominant pathogenic bacterium in children with tubercu-lous lymphadenitis .

Objective To establish the rapid pathogen identification method for HIV and Mycobac-terium tuberculosis (Mtb)co-infection and the assay for the drug susceptibility. Methods Geneprobe and 16S rDNA sequencing were used to differentiate mycobacterium species and modified microscopic observation drug susceptibility (MODS) was used for the drug susceptibility test. The above assays were compared with acid-fast smear, L-J culture and proportional drug susceptibility tests. Results (1) Thirty-four mycobacte-rial isolates were obtained from 112 samples collected from 68 HIV patients. Among these isolates, the strain species were determined by Geneprobe and 16S rDNA sequencing as the followings: 21 Mtb complex, 10 NTM including 5 M.avium complex, 2 M.gordonae, 2 M.kansasii, 1 M.colombiense, and 3 co-infection. (2) The sensitivity of Mtb to rifampicin, ethambutol, isoniazid and streptomycin were 100%, 100%, 76.2%, 90.5% respectively, while the sensitivity of NTM to rifampicin, ethambutol, isoniazid and strepto-mycin were 40%, 60%, 0%, 30% respectively. There is no significant statistic difference between the two methods, MODS and the reference standard, for the drug susceptibility test. (3) Six to eight weeks are nee-ded for the identification of the species of mycobacteria and the drug susceptibility test by using traditional method. In this study, 5-14 d, 6-15 d and 10-14 d are needed for Geneprobe, 16S rDNA sequencing, and MODS respectively. The time for the testing has been dramatically shortened. Conclusion The identifica-tion of mycobacterial species and the drug susceptibility test using clinical samples could be completed within 15 days by using combined Geneprobe, 16S rDNA sequencing and modified MODS. This combined method can be used for the pathogen identification and drug resistant test in HIV patients who are co-infected by my-cobacteria.

BACKGROUND:With the increase of aging degree, chronic wounds are increasing. There is few method to cure chronic wounds at present, and vacuum sealing drainage has been applied to the clinic.
OBJECTIVE:To investigate the effect of vacuum sealing drainage on the expression of nerve growth factor and the change of microvascular number in the human chronic wounds, and to study the mechanism of accelerating chronic wound healing.
METHODS:Ten patients with chronic wounds were included in this study, including one case of soft tissue defect of chest, one case of osteomyelitis with bone exposure, one case of large area skin defect after amputation stump, two cases of wound infection after skin avulsion injury, two cases of shinbone osteomyelitis, and three cases of postoperative wound infection. The skin around the wounds and granulation tissue within the wounds were col ected before vacuum sealing drainage treatment and at 7, 14 days after vacuum sealing drainage treatment. The nerve growth factor expressions and the microvascular number in the wounds were measured by the immunohistochemical technology. The wound healing was also observed.
RESULTS AND CONCLUSION:The nerve growth factor expressions in fibroblasts and vascular endothelial cel s at 7 and 14 days after vacuum sealing drainage treatment were significantly higher than that before treatment (P<0.001). The microvascular number was also increased after vacuum sealing drainage treatment compared with pre-treatment (P<0.05-0.01). The vacuum sealing drainage treatment reduced necrotic tissue and purulent secretion within the wounds, as wel as swel ing around the wounds. Simultaneously, granulation tissue gradual y became fine granules, bright red, and bleeding easily, thus it is suitable for promoting wound healing.

Objective To investigate the correlation between the mutation of pncA gene and the susceptibility to pyrazinamide ( PZA) in Mycobacterium tuberculosis complex ( MTBC) strains and to analyze the mutation of panD and rpsA genes in wild type isolates without pncA gene mutation.Methods The sus-ceptibilities of 108 MTBC strains to first-line drugs including PZA were detected by using the MGIT 960 TB system.PCR was performed to amplify the 16S rDNA and pncA, panD and rpsA genes.The PCR products were analyzed by DNA sequencing analysis .Results Among the 78 multidrug-resistant MTBC strains , 47 isolates (60%) were resistant to PZA.Four out of 30 (13%) strains that were sensitive to ethambutol , iso-niazid, rifampicin and streptomycin (EIRS) were resistant to PZA.The drug-resistant MTBC strains showed higher resistance rate to PZA than that of the EIRS sensitive strains .There were 49 ( 96%) PZA-resistant isolates and 4 (7%) PZA-sensitive isolates occurred pncA gene mutation.Most of the pncA gene mutations in the genomes of PZA-resistant strains were base substitution mutation , especially the His57Asp substitu-tion.The pncA gene mutations centralized in the regions of 160-169, 203-289, 309-396 and 413-467.Seven novel mutation sites of pncA gene were observed including T175C, C188A, G insertion at 68, AGC insertion at 235, C insertion at 339, CC insertion at 392 and GT deletion at 395.The mutation sites founded in the genomes of PZA-sensitive strains were different from those of the PZA-resistant strains .No mutation of the pncA gene and the upstream regulatory sequence was found in two PZA-resistant strains , NJ44 and NJ108 . The sequence analysis of panD and rpsA gene showed that the NJ 108 strain had panD gene mutation at G419A, but no mutation was detected in the NJ 44 strain.Conclusion The multidrug-resistant MTBC strains showed higher resistance rate to PZA .The pncA gene mutation was common in PZA-resistant MTB strains and the panD gene mutation was also worthy of attention .

Objective To explore the risk factors of skeletal related events (SREs) in non small cell lung cancer with bone metastases and its effect on the prognosis .Methods Totally 223 cases of NSCLC patients with bone metastasis were retrospective studied from January 2010 to December 2012 in our hospital .The clinical features ,predictive factors for SREs were analysed by sin‐gle factor and multifactor analysis .Results Among 223 cases of NSCLC patients with bone metastasis ,119 cases occured with SREs(53 .4% ) .Univariate analysis showed that the occurrence of SREs in female ,no smoker ,adenocarcinoma ,solitary bone metas‐tasis lesions were less than the male ,smoker non‐adenocarcinoma ,and multiple bone metastases (P<0 .05) ,but the rost without statistically significant(P>0 .05) .The multivariate analysis revealed only multiple bone metastases was an independent risk factor for SREs .The median survival time of the NSCLC patients with bone metastasis was 15 .3 months .Moreover ,survival analysis showed that SREs had no statistical significance on the prognosis of bone metastasis in NSCLC patients (P>0 .05) .Conclusion The female ,adenocarcinoma ,smoking history ,solitary bone metastasis lesions occurred in patients with lower risk SREs .Multiple bone metastasis is an independent risk factor for SREs ,attention should be paid to monitoring and prevention .

ObjectiveTo analyze the characterstics of phenotype and genotype of multidrug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) by molecular line probe assay and liquid culture with MGIT960.MethodsGenoType MTBDR Kits were used for identifying the types of the first-line and second-line antituberculosis drug resistant genes partly and BD MGIT960 was used for detecting the chug susceptibility.Results( 1 ) Out of 94 MDR-TB strains,the rate of drug resistant to EMB,AMK,OFX and MFX by BD MGIT960 assay were 36.2%,17.0%,54.3% and 55.3%,respectively.Among these isolates,13 were extensively drug resistant tuberculosis (XDR-TB).(2) Compared with MGIT960,the concordance rate of GenoType MTBDRplus was 86.2% and 95.7% respectively.Taking MGIT960 results as reference,the sensitivity of GenoType MTBDRsl detecting the susceptibility of EMB,AMK,OFX and MFX to 94 isolates were 47.1%,81.3%,94.1%,94.2%,respectively.The specificity were 75.0%,98.7%,90.7%,92.9%,respectively.(3) Among the rpoB mutation categories,S531L accounts for most.MTB resistant to IFN caused by the mutation of katG chiefly and the S315T1 was in the majority.The gyrA mutation sites located at the ninety-fourth codon most.Out of 94 strains,23 were mixed with 2 kindsof Mycobacterium tuberculosis at least and 7 were undetectable mutations.Conclusion Among the M/XDR-TB,the strains resistant to INH,RFP,AMK,OFX and MFX were caused most by the mutation of katG,rpoB,rrs and gyrA,respectively.The relationship between EMB and embB was not so clear relatively.As a fast detecting drug susceptibility test kit,GenoType MTBDR possess good sensitivity and specificity.So,it could be as an assistant method to guide the therapy on clinic.

Objective:To investigate the effect of immune checkpoint inhibitors combined with concurrent chemotherapy for non-small cell lung cancer (NSCLC) and the effect of this regimen on serum levels of tumor marker and immune cells of patients.Methods:The clinical data of 60 NSCLC patients in Xuzhou Cancer Hospital from February 2020 to February 2022 were retrospectively analyzed, and they were divided into chemotherapy combined with immune checkpoint inhibitor treatment group (combination treatment group) and conventional chemotherapy group by treatment methods, with 30 cases in each group. Before treatment and 6 weeks after treatment, the patients' serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), vascular endothelial growth factor (VEGF), cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) were detected by chemiluminescence immunoassay, and the levels of serum tumorous M2 pyruvate kinase (TuM2-PK) and fatty acid synthase (FAS) were detected by double-antibody sandwich enzyme-linked immunosorbent assay. The levels of T cell subsets were measured by flow cytometry, and the quality of life of patients was evaluated according to the World Health Organization quality of life scale brief version (WHOQOL-BREF). The clinical efficacy, tumor markers levels, immune cells levels, quality of life and adverse reactions were compared between the two groups.Results:The overall effective rate of patients in the combination treatment group was 46.67% (14/30), which was higher than 20.00% (6/30) in the conventional chemotherapy group ( χ2 = 4.80, P = 0.029). The differences in serum CEA, CA125, VEGF, CYFRA21-1, TuM2-PK, FAS levels and the proportions of CD3 +, CD4 +, CD8 + T cells and WHOQOL-BREF scores between the two groups before treatment were not statistically significant (all P > 0.05); the levels of CEA, CA125, VEGF, CYFRA21-1, TuM2-PK, FAS and the proportion of CD8 + T cells at 6 weeks after treatment were lower than those before treatment in both groups (all P < 0.05), and the proportions of CD3 + and CD4 + T cells and WHOQOL-BREF scores were higher than those before treatment (all P < 0.05); the levels CEA, CA125, VEGF, CYFRA21-1, TuM2-PK and the proportions of CD8 + T cells in the combination treatment group at 6 weeks after treatment were higher than those in the conventional chemotherapy group at 6 weeks after treatment (all P < 0.001), and the proportions of CD3 + and CD4 + T cells and WHOQOL-BREF scores were higher than those in the conventional chemotherapy group at 6 weeks after treatment (all P < 0.05). The differences in the incidence of gastrointestinal reactions, alopecia, leukopenia, thrombocytopenia, and liver and kidney function impairment between the two groups were not statistically significant (all P > 0.05). Conclusions:Immune checkpoint inhibitors combined with chemotherapy in NSCLC patients are more effective than conventional chemotherapy, and the combined treatment can more effectively reduce the serum tumor marker levels of patients and enhance the anti-tumor immune effect, with the adverse reactions comparable to conventional chemotherapy.

OBJECTIVE: To carry out mutation analysis for a Chinese family affected with autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS). METHODS: Whole exome sequencing (WES) was used to screen potential mutations within genomic DNA extracted from the proband. Suspected mutation was validated by combining clinical data and results of Sanger sequencing. RESULTS: A homozygous deletional mutation c.3665_3675delGTGCTGTCTTA (p.S1222fs) was found in the proband, for which her parents were both heterozygous carriers. CONCLUSION WES is capable of detecting mutation underlying this disorder and facilitating genetic counseling and prenatal diagnosis for the affected family. A novel pathogenic mutation of the SACS gene was discovered.
Female ; Genes, Recessive ; Heat-Shock Proteins ; genetics ; Humans ; Muscle Spasticity ; Mutation ; Spinocerebellar Ataxias ; congenital

Objective:To understand cytomegalovirus (CMV) infection status in hospitalized preterm infants who were fed by their own mother's frozen breast milk.Methods:This retrospective study enrolled breastfed neonates with gestational age less than 32 weeks or birth weight less than 1 500 g who were born and admitted to Gynecology and Obstetrics Hospital of Fudan University from January 2018 to December 2020. Clinical data of the babies and their mothers were collected and analyzed, including CMV DNA results of breast milk and urine samples of the subjects by fluorescence quantitative polymerase chain reaction. Chi-square test (or Fisher's exact probability test), two independent samples t test, and Mann-Whitney U test were used for the statistical analysis. Results:A total of 94 parturients and their 103 premature infants (including nine pairs of twins) were included. CMV DNA of breast milk was noted for positive in 75 cases (including eight pairs of twins) and for negative in 28 cases (including one pair of twins). Out of the 75 preterm infants born to mothers with positive CMV DNA breast milk, 67 (including eight pairs of twins) were switched to frozen breast milk (－20 ℃ for 72 h), and six of them were infected by CMV(9.0%) without any treatment. All of the 103 infants were divided into two groups: the frozen milk fed group ( n=67) or fresh milk fed group ( n=36). In the frozen milk fed group, the CMV DNA was mainly detected during 2-8 weeks postpartum with copy number reaching the peak at 8th week. And those infants in the frozen milk fed group, whose mother's breast milk CMV DNA was positive, was further divided into CMV infected ( n=6) or CMV non-infected groups ( n=61) according to the urine test results. Moreover, compared with the non-infected group, the average [22.7(3.0-95.7)×10 3 copies/ml vs 5.0(0.5-89.5)×10 3 copies/ml, Z=－2.218) and the highest[45.9(5.9-261.0)×10 3 copies/ml vs 9.8(1.2-766.0)×10 3 copies/ml, Z=－2.218] copy number of CMV DNA in the breast milk were higher in the CMV infected group (both P<0.05). The incidence of feeding intolerance[37.3% (25/67) vs 50.0% (18/36), χ2=1.550], neonatal necrotizing enterocolitis [0.7% (1/67) vs 0.0% (0/36)], bronchopulmonary dysplasia [28.4% (19/67) vs 27.8% (10/36), χ2=0.004], retinopathy of prematurity [20.9% (14/67) vs 8.3%(3/36), χ2=2.682], and late-onset sepsis [22.4% (15/67) and 30.6% (11/36), χ2=0.828] did not differ significantly between the frozen or fresh milk fed groups (all P>0.05). Conclusions:The incidence of breast milk-related CMV infection in those fed with frozen breast milk was low and does not increase the without increasing risks of related complications or leading to obvious clinical manifestations after infection. For preterm infants with gestational age <32 weeks or birth weight <1 500 g, frozen breast milk can be an alternative for mothers with CMV DNA positive breast milk.