Objective To analyze the characteristics of drug resistance in patients with disseminated pulmonary tuberculosis,and provide references for the clinical development of individualized treatment plans.Methods A retrospective analysis was conducted on 71 hospitalized patients with disseminated pulmonary tuberculosis treated at the Beijing Chest Hospital,Capital Medical University from January 2015 to January 2024.The susceptibility of Mycobacterium tuberculosis from these patients to 16 anti-tuberculosis drugs was detected using the microplate method for mycobacterial drug susceptibility testing.The study analyzed the culture results of Mycobacterium tuberculosis,drug susceptibility test results,initial treatment or re-treatment status,drug resistance,and differences in drug resistance types between initial and re-treated patients.Results Among the 71 patients with disseminated pulmonary tuberculosis,there were 51 males(71.8%),and 58 cases(81.7%)of acute disseminated pulmonary tuberculosis,with an overall drug resistance rate to 16 anti-tuberculosis drugs of 38.0%.There was no statistically significant difference in the total drug resistance rate between re-treated patients and those undergoing initial treatment[52.2%(12/23)vs.31.3%(15/48),P=0.089].The top 7 drugs to which patients were resistant were streptomycin(Sm)and isoniazid(INH)with 13 cases each(18.3%),rifapentine(Rft)and isoniazid aminosalicylate(Pa)with 10 cases each(14.1%),rifampicin(RFP),rifabutin(Rfb),and capreomycin(Cm)with 9 cases each(12.7%).The top 6 drugs to which initially treated patients were resistant were Cm with 6 cases(12.5%),Sm with 5 cases(10.4%),Pa with 4 cases(8.3%),INH,clarithromycin(Clr),and p-aminosalicylic acid(PAS)with 3 cases each(6.3%).The top 7 drugs to which re-treated patients were resistant were INH with 10 cases(43.5%),Sm,RFP,and Rft with 8 cases each(34.8%),Rfb with 7 cases(30.4%),Pa and levofloxacin(Lfx)with 6 cases each(26.1%).The overall mono-resistance rate,poly-drug resistance rate,and multidrug-resistant rate to 16 anti-tuberculosis drugs were 9.9%,7.0%,and 11.3%,respectively;all mono-resistance patients were initially treated;there was no statistically significant difference in the poly-drug resistance rate between re-treated and initially treated patients(13.0%vs.4.2%,P=0.591),but the multidrug-resistant rate was significantly higher in re-treated patients(30.4%vs.2.1%,P=0.002).Conclusion Drug resistance in disseminated pulmonary tuberculosis is severe.Clinical physicians can develop personalized anti-tuberculosis treatment plans based on drug susceptibility test results.

OBJECTIVE: To derive the Chinese medicine (CM) syndrome classification and subgroup syndrome characteristics of ischemic stroke patients. METHODS: By extracting the CM clinical electronic medical records (EMRs) of 7,170 hospitalized patients with ischemic stroke from 2016 to 2018 at Weifang Hospital of Traditional Chinese Medicine, Shandong Province, China, a patient similarity network (PSN) was constructed based on the symptomatic phenotype of the patients. Thereafter the efficient community detection method BGLL was used to identify subgroups of patients. Finally, subgroups with a large number of cases were selected to analyze the specific manifestations of clinical symptoms and CM syndromes in each subgroup. RESULTS: Seven main subgroups of patients with specific symptom characteristics were identified, including M3, M2, M1, M5, M0, M29 and M4. M3 and M0 subgroups had prominent posterior circulatory symptoms, while M3 was associated with autonomic disorders, and M4 manifested as anxiety; M2 and M4 had motor and motor coordination disorders; M1 had sensory disorders; M5 had more obvious lung infections; M29 had a disorder of consciousness. The specificity of CM syndromes of each subgroup was as follows. M3, M2, M1, M0, M29 and M4 all had the same syndrome as wind phlegm pattern; M3 and M0 both showed hyperactivity of Gan (Liver) yang pattern; M2 and M29 had similar syndromes, which corresponded to intertwined phlegm and blood stasis pattern and phlegm-stasis obstructing meridians pattern, respectively. The manifestations of CM syndromes often appeared in a combination of 2 or more syndrome elements. The most common combination of these 7 subgroups was wind-phlegm. The 7 subgroups of CM syndrome elements were specifically manifested as pathogenic wind, pathogenic phlegm, and deficiency pathogens. CONCLUSIONS There were 7 main symptom similarity-based subgroups in ischemic stroke patients, and their specific characteristics were obvious. The main syndromes were wind phlegm pattern and hyperactivity of Gan yang pattern.
Humans ; Syndrome ; Ischemic Stroke ; Medicine, Chinese Traditional ; Liver ; Phenotype

OBJECTIVE: To investigate whether cell preservation solution can prolong the survival time of leukemia cells and increase the survival rate, so as to improve the detection rate of central nervous system leukemia. METHODS: Kasumi cells were added into cerebrospinal fluid (CSF) supernatant with or without cell preservation solution to compare cell viability and biological characteristics at different time point. Wright Giemsa staining was used to compare cell morphology; cell counting, CCK-8 method, and trypan blue staining were used to compare the cell number, and flow cytometry was used to compare the cell viability. The expression of AML-ETO tumor fusion gene was detected by fluorescence quantitative RT-PCR. RESULTS: At different time points (8 h and 24 h), the survival, molecular biological characteristics and RT-PCR result of the cells in CSF with cell preservation solution were significantly better than those in normal cerebrospinal fluid. CONCLUSION Cell preservation solution can effectively improve the survival time and survival rate of leukemic cells, thereby increase the detection rate of CNS leukemia.
Central Nervous System Neoplasms ; Core Binding Factor Alpha 2 Subunit ; Humans ; Leukemia ; RUNX1 Translocation Partner 1 Protein

OBJECTIVETo establish allo-transplantation model by using mRFP⁺ to eGFP⁺ transgenic mice and to observe the distribution of donor cells and donor-recipient cellular interaction in the bone marrow after semi-solid decalcification (SSD).

METHODSAfter myeloablative irradiation, C57BL/6 female eGFP⁺ transgenic mice were infused with (5 × 10⁶) bone marrow cells from FVB male donor mice through tail vein. The control group was infused with PBS. Then the general conditions, engraftment level, hematopoietic recovery, incidence of GVHD and survival of recipients were evaluated after transplantation. In the recovery process, SSD was used to treat the femora before observing the cells distribution, morphology and interaction by confocal microscopy directly or after making frozen section.

RESULTSWBC of recipient eGFP⁺ mice was recovered on (20 ± 3.07) d, (93.94 ± 1.59)% in peripheral cells were RFP⁺ cells (n = 10), GVHD happened in 4 of 10 mice within 1 month. During SSD, the hard components were replaced gradually and RFP⁺ cells could be seen mainly in the bone trabecula and surrounded by eGFP⁺ cells under confocal microscope, their interactions could be further observed clearly in bone marrow microenvironment in three-dimensional reconstruction.

CONCLUSIONThe double fluorescent allo-transplantation mouse model successfully established, by means of our novel protocol named SSD, the donor and recipient cell location and their interaction can be visually observed, which provides the basis for clinical studies on the distribution and homing of donor cells, and some related explorations after transplantation.

Animals ; Bone Marrow Cells ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Graft vs Host Disease ; Green Fluorescent Proteins ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Transgenic

BACKGROUNDInfantile hemangioma (IH) is the most common benign tumor in children with prevalence in the face and neck. Various treatment options including oral propranolol have been described for IH, but the mechanism of drugs remains enigmatic. The aim of this study was to investigate the pathogenesis and establish a reliable in vivo model of IH which can provide platform for drug exploration.

METHODSStem cells from the proliferating hemangiomas (HemSCs) were isolated by CD133-tagged immunomagnetic beads. Their phenotype and angiogenic property were investigated by flow cytometry, culturing on Matrigel, real-time polymerase chain reaction (PCR), immunofluorescent staining and injection into BALB/c-nu mice.

RESULTSHemSCs had robust ability of proliferating and cloning. The time of cells doubling in proliferative phase was 16 hours. Flow cytometry showed that HemSCs expressed mesenchymal markers CD29, CD44, but not endothelial/hematopoietic marker of CD34 and hematopoietic marker CD45. The expression of CD105 was much lower than that of the reported hemangioma derived or normal mesenchymal stem cell (MSC). Real-time PCR showed that the mRNA levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and matrix metalloproteinase-1 (MMP-1) of HemSCs were higher than that of neonatal human dermal fibroblasts (NHDFs) and human umbilical vein endothelial cells (HUVECs). After HemSCs were cultured on Matrigel in vitro, they formed tube-like structure in a short time (16 hours) and differentiated into endothelial cells in 7 days. After 1 - 2 weeks of implantation into immunodeficient mice, HemSCs generated glucose transporter 1 positive blood vessels. When co-injected with HUVECs, the vascularization of HemSCs was greatly enhanced. However, the single implantation of HUVECs hardly formed blood vessels in BALB/c-nu mice (P < 0.05).

CONCLUSIONSHemSCs may be some kinds of primitive mesoderm derived stem cells with powerful angiogenic ability, which can recapitulate human hemangioma by co-injecting into immunodeficient mice with HUVECs.

AC133 Antigen ; Animals ; Antigens, CD ; analysis ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Collagen ; Disease Models, Animal ; Drug Combinations ; Glycoproteins ; analysis ; Hemangioma ; pathology ; Humans ; Laminin ; Male ; Mice ; Mice, Inbred BALB C ; Neoplastic Stem Cells ; chemistry ; pathology ; Peptides ; analysis ; Proteoglycans ; Vascular Endothelial Growth Factor A ; physiology

Background Infantile hemangioma (IH) is the most common benign tumor in children with prevalence in the face and neck.Various treatment options including oral propranolol have been described for IH,but the mechanism of drugs remains enigmatic.The aim of this study was to investigate the pathogenesis and establish a reliable in vivo model of IH which can provide platform for drug exploration.Methods Stem cells from the proliferating hemangiomas (HemSCs) were isolated by CD133-tagged immunomagnetic beads.Their phenotype and angiogenic property were investigated by flow cytometry,culturing on Matrigel,real-time polymerase chain reaction (PCR),immunofluorescent staining and injection into BALB/c-nu mice.Results HemSCs had robust ability of proliferating and cloning.The time of cells doubling in proliferative phase was 16 hours.Flow cytometry showed that HemSCs expressed mesenchymal markers CD29,CD44,but not endothelial/hematopoietic marker of CD34 and hematopoietic marker CD45.The expression of CD105 was much lower than that of the reported hemangioma derived or normal mesenchymal stem cell (MSC).Real-time PCR showed that the mRNA levels of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF) and matrix metalloproteinase-1 (MMP-1) of HemSCs were higher than that of neonatal human dermal fibroblasts (NHDFs) and human umbilical vein endothelial cells (HUVECs).After HemSCs were cultured on Matrigel in vitro,they formed tube-like structure in a short time (16 hours) and differentiated into endothelial cells in 7 days.After 1-2 weeks of implantation into immunodeficient mice,HemSCs generated glucose transporter 1 positive blood vessels.When co-injected with HUVECs,the vascularization of HemSCs was greatly enhanced.However,the single implantation of HUVECs hardly formed blood vessels in BALB/c-nu mice (P ＜0.05).Conclusions HemSCs may be some kinds of primitive mesoderm derived stem cells with powerful angiogenic ability,which can recapitulate human hemangioma by co-injecting into immunodeficient mice with HUVECs.

Objective To evaluate the short-term efficacy of lamivudine versus entecavir for patients with HBeAg-negative acute-on-chronic liver failure (ACLF) with different pretreatment liver failure degrees.MethodsA total of patients with HBeAg-negative ACLF were enrolled into this retrospective study.Seventy-two cases were treated with lamivudine 100 mg daily,while 93 cases were treated with entecavir 0.5 mg daily.Biochemical items,model for end-stage liver disease (MELD)score,hepatitis B virus (HBV) DNA level and mortality were observed.The efficacies of the two drugs were analyzed in patients with different degrees of liver failure.The comparison of rates was done using chi-square test and the measurement data were compared by t test.ResultsAmong the patients with pretreatment MELD scores above 30,the post-treatment HBV DNA levels in lamivudine group and entecavir group were (3.6 ± 1.1) lg copy/mL and (3.7 ± 1.4) lg copy/mL,respectively (t=0.181,P=0.859) and the mortalities were 92.0％ and 91.8％,respectively (χ2 =0.002,P=0.680).For the patients with pretreatment MELD scores from 23 to 30,the post-treatment HBV DNA levels in two groups were (3.2± 1.1) lg copy/mL and (3.2±2.3) lg copy/mL,respectively (t=0.760,P=0.455) and the mortalities were 42.9％,54.1％,respectively (χ2 =0.799,P=0.455).In patients with pretreatment MELD scores below 23,the post-treatment HBV DNA levels in two groups were (3.1±1.0) lg copy/mL and (2.8±1.5) lg copy/mL,respectively (t=-0.740,P=0.464) and the mortalities were 3/19 and 6.3％,respectively (χ2=1.227,P=0.455).In lamivudine group,the mortalities were significantly different among patients with three different ranges of pretreatment MELD scores (χ2 =26.967,P =0.000).The similar differences were also found in entecavir group (χ2 =41.260,P=0.000).ConclusionsAmong treatment na?ve patients with HBeAg-negative ACLF,the short-term efficacy of lamivudine versus entecavir is equal if the degree of pretreatment liver failure is similar.Meanwhile,the degrees of pretreatment liver failure significantly affects the outcome of the treatment.

OBJECTIVETo explore the relationship between sexual hormones in semen and germ cell apoptosis in male population.

METHODSSixty-six infertile patients and thirty fertile males were selected randomly. The levels of folicle stimulating hormone ( FSH), prolactin (PRL), luteinizing hormone (LH), and testosterone (T) in semen were measured by ELISA. Terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL) was used for the detection of germ cell apoptosis.

RESULTSThe levels of FSH, LH, PRL, T in thirty fertile men were (1.63 +/- 0.15) U/L, (2.18 +/- 0.21) U/L, (6.34 +/- 0.30) nmol/L, (1.85 +/- 0.11) nmol/L, respectively, and germ cell apoptosis rate was (4.61 +/- 1.23)%. FSH, LH, PRL, T levels in infertile group were (1.25 +/- 0.18) U/L, (1.76 +/- 0.32) U/L, (5.86 +/- 0.13) nmol/l, (1.45 +/- 0.13) nmol/, respectively, and germ cell apoptosis rate was (18.36 +/- 2.04)%. There were significant differences in all parameters between infertile group and fertile group. The levels of FSH, LH, PRL, T were negatively correlated with germ cell apoptosis rates( r = -0.88, -0.93, -0.90, -0.98). The volume of apoptotic germ cell decreased, and chromatin was compacted to form cell-membrane blebs and apoptotic bodies.

CONCLUSIONLow concentration of sexual hormones may increase the apoptosis of germ cells, which can induce male infertility.

Adult ; Apoptosis ; Case-Control Studies ; Follicle Stimulating Hormone ; metabolism ; Germ Cells ; pathology ; Gonadal Steroid Hormones ; metabolism ; Humans ; Infertility, Male ; metabolism ; pathology ; Luteinizing Hormone ; metabolism ; Male ; Prolactin ; metabolism ; Semen ; metabolism ; Testosterone ; metabolism

OBJECTIVETo explore the effect of nitric oxide (NO) on human sperm capacitation and acrosome reaction (AR).

METHODSDifferent concentrations of sodium nitroprusside (SNP) were added to the sperm suspension from 48 healthy fertile men, and the suspension was incubated in 1 x Earle at 37 degrees C for 1 hour. Progesterone was used to induce AR for 15, 30, 45 and 60 min, and then acid phosphatase (ACP) activity in the suspension before and after capacitation and at different time of AR was measured by p-nitrophenyl sodium phosphate assay. In the meantime, sperm motile parameters were assayed by CASA to observe sperm capacitation and AR.

RESULTSACP activity and sperm motile parameters increased in the 50 approximately 100 nmol/L NO concentration group, showed no significant variation in the 150 approximately 200 nmol/L group, and decreased in the 250 approximately 300 nmol/L group.

CONCLUSIONNO can facilitate sperm capacitation, AR and sperm motile parameters in low concentration and suppress them in high concentration. ACP activity assay of sperm is an objective and reliable method to evaluate sperm capacitation and AR in whole sperm population.

Acid Phosphatase ; metabolism ; Acrosome Reaction ; drug effects ; physiology ; Adult ; Dose-Response Relationship, Drug ; Humans ; Male ; Nitric Oxide ; physiology ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Sperm Capacitation ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; enzymology

Objective To observe the morphologic changes of peripheral blood lupus cell in patients with systemic lupus erythematosus and investigate its relationship with disease activity in systemic lupus ery- thematosus.Methods Modified classical blood clotting method to observe the morphological changes of pe- ripheral blood lupus cells in 80 cases with systemic lupus erythematosus.Fifty cases were in active stage,and 30 cases were in stable stage.Comparison of serum autoantibody,complement and SLEDAI were also carried out.Results There was significant association between lupus cells in special morphous and autoantibody, such as anti-double-stranded DNA antibody,anti-nucleosome antibodies,complement C3、C4 and SLEDAI(r= 0.588,P=0.056:r=0.759,P=0.135;r=-0.648,P=0.058;r=-0.589,P=0.057,r=0.686,P