Objective:To investigate the clinical characteristics, treatment and prognosis of newly-treated patients with primary central nervous system lymphoma (PCNSL).Methods:Clinical data of 117 newly-treated PCNSL patients who were admitted to the First Hospital of Jilin University, the Fifth Medical Center of Chinese PLA General Hospital, Harbin Medical University Cancer Hospital, and Cancer Hospital of Chinese Academy of Medical Sciences & Peking Union Medical College from August 2009 to February 2018 were retrospectively analyzed. The patients' age, sex, Eastern Cooperative Oncology Group (ECOG) physical status (PS) score, pathological type, involvement of deep brain tissue, number of lesions, cerebrospinal fluid protein concentration, International Extranodal Lymphoma Study Group (IELSG) score, Memorial Sloan Kettering Cancer Center (MSKCC) score, treatment strategy, and response after the first-line therapy were analyzed using univariate and multivariate Cox proportional hazards models to identify the independent influencing factors for progression-free survival (PFS) and overall survival (OS) of PCNSL patients. Kaplan-Meier method was used for survival analysis.Results:In 117 newly-treated PCNSL patients, 59 cases (50.4%) presented with increased intracranial pressure or focal neurological symptoms at diagnosis; there were 65 cases (55.6%) with single lesions and 52 cases (44.4%) with multiple lesions; 1 patient (0.9%) had lymphoma of T-cell origin, and 116 cases (99.1%) had diffuse large B-cell lymphoma (DLBCL). Among 95 evaluable patients, 41 patients (43.2%) achieved complete remission (CR), 20 patients (21.1%) achieved partial remission (PR), 16 patients (16.8%) achieved stable disease (SD), and 18 patients (18.9%) had progressive disease (PD). In 117 patients with median follow-up of 66.0 months (95% CI 57.9-74.1 months), the median PFS and OS were 17.4 months (95% CI 11.5-23.3 months) and 45.6 months (95% CI 20.1-71.1 months), respectively. The 2-, 3- and 5-year PFS rates were 41.2%, 28.6% and 19.3%, and OS rates were 63.7%, 52.4% and 46.3%, respectively. Univariate Cox regression analysis showed that baseline high-risk MSKCC score group was an adverse prognostic factor for PFS ( P = 0.037), and the first-line chemotherapy with ≥4 cycles of high-dose methotrexate (HDMTX), HDMTX in combination with rituximab, ≥4 cycles of rituximab in combination with HDMTX, and achieving CR or ≥PR after the first-line treatment reduced the risk of disease progression and prolonged the PFS time (all P <0.01); age >60 years old, ECOG-PS score of 2-4 points, elevated cerebrospinal fluid protein concentration, high-risk IELSG score, and high-risk MSKCC score were adverse prognostic factors for OS, and ≥4 cycles of HDMTX and achieving CR or ≥PR after the first-line treatment were favorable factors for OS. Multivariate Cox regression analysis verified that rituximab in combination with HDMTX (yes vs. no: HR = 0.349, 95% CI 0.133-0.912, P = 0.032) and achieving ≥PR after the first-line chemotherapy (yes vs. no: HR = 0.028, 95% CI 0.004-0.195, P < 0.001) were independent favorable factors for PFS; age >60 years old (>60 years old vs. ≤60 years old: HR = 10.878, 95% CI 1.807-65.488, P = 0.009) was independent unfavorable factor for OS, while ≥4 cycles of HDMTX treatment (≥4 cycles vs. <4 cycles: HR = 0.225, 95% CI 0.053-0.947, P = 0.042) was independent favorable factor for OS. Conclusions:The older the PCNSL patients at initial treatment, the worse the prognosis. Intensive and continuous treatment for achieving deeper remission may be the key for improving the outcome of PCNSL patients.

Objective:To study the clinical application of metagenomic next-generation sequencing (mNGS) on pathogen diagnosis and clinical management in neonatal sepsis.Methods:From January 2019 to February 2021, neonates suspected of sepsis receiving both mNGS and blood culture in our hospital were retrospectively reviewed. The effects of mNGS on pathogen detection, antibiotic choice and clinical management were analyzed.Results:A total of 35 neonates were included. Sepsis was confirmed in 24 neonates. 12 neonates had positive microbiology results (both mNGS and blood culture were positive in 3 cases, only mNGS positive in 8 cases and only blood culture positive in 1 case). mNGS showed higher positive rate than blood culture in patients with neonatal sepsis (45.8% vs. 16.7%, P=0.049). The concordance of mNGS and blood culture was 74.3%. 21 cases (60.0%) had antibiotics before testing and the positive rate showed no significant difference between mNGS and blood culture [19.0% (4/21) vs. 4.8% (1/21), P=0.343] in these patients. 14 cases didn't receive antibiotics before testing and the positive rate also showed no significant difference between mNGS and blood culture [28.6%(4/14) vs.21.4% (3/14), P=1.000]. 8.6% patients changed clinical treatment based on the mNGS results. Conclusions:mNGS increases pathogen detection in neonatal sepsis and is well consistent with blood culture. mNGS is useful in clinical practice.

Benign childhood epilepsy with centrotemporal spikes (BECT) is the most common childhood idiopathic focal epilepsy and used to be considered as one of the childhood epilepsy syndromes with the best prognosis. In recent years, more and more studies have found that patients with BECT have varying degrees of cognitive dysfunction. This article reviews some of research progresses in the clinical features, neuropsychology and neuroimaging of patients with BECT, in order to explore the effects of seizures, age of onset, interictal discharges, changes of brain structure and functional network, as well as antiepileptic drugs on cognitive function in patients with BECT.

Objective:To investigate the role of cyclo-oxygenase-2 (COX-2) in breast cancer metastasis and its possible mechanism. Methods: A total of 45 cases of primary breast cancer tissues and brain metastatic breast cancer tissues were collected from patients, who underwent mastectomy in Yunnan Cancer Hospital from October 2015 to April 2018, including 30 cases of primary lesions and 15 cases of brain metastasis. qPCR was used to detect the expression of COX-2 in breast cancer tissues and brain metastatic breast cancer tissues. Recombinant viruses with COX-2 over-expression (LV6-COX2) or COX-2 knockdown (LV3-COX2 shRNA1, LV3-COX2 shRNA2) were transfected into human breast cancer MDA-MB-231 cells; After obtaining the stable expression cell lines, the effect of COX-2 expression on the proliferation of MDA-MB-231 cells was detected by CCK-8, and the effects of COX-2 expression on the migration and invasion of MDA-MB-231 cells were detected by scratch test and Transwell assay, respectively. The mRNAand protein expressions of COX-2 in each group were examined by qPCR and WB, respectively. The effect of COX-2 expression on the expression of EMT-related genes in MDA-MB-231 cells was analyzed by qPCR. Results: The expression of COX-2 in tissues of patients with brain metastases was significantly higher than that in patients with primary breast cancer tissues (P<0.01), and it was correlated with tumor TMN stage in breast cancer patients. MDA-MB-231 cell lines with stable COX-2 over-expression/knockout were successfully constructed. Over-expression of COX-2 promoted the migration and invasion of MDA-MB-231 cells (all P<0.01), and significantly increased the expressions of MMP2, MMP1, N-cadherin and vimentin (all P<0.01), but exerted insignificant effect on cell proliferation. The effect of COX-2 silence exerted the opposite effect and promoted cell proliferation (P<0.05). Conclusion: COX-2 is highly expressed in brain metastatic breast cancer tissues, which may promote the migration and invasion of breast cancer MDA-MB-231 cells by regulating EMT processes.

Objective:To study the inhibitory effect of deoxyschizandrin on the growth of brain glioma C6 cells, and to explore its mechanism.Methods:The rat glioma C6 cells were cultured and divided into control group,50, 100,and 200 mg·L-1 deoxyschizandrin groups.The proliferation rates of C6 cells were examined by MTT assay;the changes of cell cycles were examined by flow cytometry;the expression levels of CyclinD1,Bax,Bcl-2 and Caspase-3 proteins in supernant were detected by ELISA assay. Results:Compared with control group, the proliferation rates at 24 and 48 h in 50,100,and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P <0.01),and the proliferation rates at 72 h in 100 and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P < 0.05 or P < 0.01 ). Compared with control group, the percentage of cells at SubG1 phase in 200 mg·L-1 deoxyschizandrin group was increased (P < 0.05 ), and the percentage of cells at S phase was decreased (P <0.05).Compared with control group,the expression levels of CyclinD1 in 100 and 200 mg· L-1 deoxyschizandrin groups were decreased (P < 0.01 );the expression levels of Bax protein in deoxyschizandrin groups were increased (P < 0.05 or P < 0.01 ), and the expression level of Bcl-2 protein in 200 mg · L-1 deoxyschizandrin group was decreased (P < 0.01 ), and the Bax/Bcl-2 value in deoxyschizandrin groups were increased (P < 0.01 ); the expression level of Caspase-3 protein in 200 mg · L-1 deoxyschizandrin group was increased (P < 0.01 ).Conclusion:Deoxyschizandrin could inhibit the growth of glioma cells through down-regulating the expression levels of CyclinD1 protein and up-regulating the expression levels apoptotic factors Bax and Bcl-2.

Objective To explore the relations of clinical phenotypes of type Ⅰ-ⅣV spinal muscular atrophy (SMA) with neural electrophysiological features and survival motor neuron (SMN)gene.Methods A total of 85 patients with SMA,including 46 with infantile form in which 19 of type Ⅰ and 27 of type Ⅱ,24 with juvenile form (type Ⅲ),and 15 with adult form (type ⅣV),were involved in this clinical study.Their clinical data were analyzed.The neural conduction,needle electromyography (EMG)and SMN1 gene deletion were analyzed.Results There existed different clinical features among patients who suffered from type Ⅰ to type ⅣV SMA.However,the major clinical features of SMA were displayed by progressively aggravating of flaccid paralysis in muscles of the four limbs,and the younger of the patients,the more serious of the clinical manifestations.EMG exhibited neurogenic lesion in all 43 SMA patients,33 patients presented generalized neurogenic lesions,and the abnormal degree of muscles in lower limbs was more severe than that of upper limbs,and the proximal muscles was more severe than that of the distal ones.The abnormal rate of spontaneous potential,weak contraction with raise potential and amplitude of compound motor active potential in adult and juvenile SMA were significantly lower than those in infantile SMA.SMN1 gene exon 7 and 8 were detected in all 85 patients with SMA.A total of 61 patients were found with deletion of exon 7 and/or 8 in SMN1 gene.Infantile SMA patients enjoyed 95.7％ (44/46) detection rate,juvenile SMA patients enjoyed 70.8％ (17/24) detection rate;no adult SMA patients were found with deletion ofexon 7 and/or 8 in SMN1 gene.Conclusions The more serious of clinical manifestations in SMA patients,the higher abnormality rote in electrophysiological tests.The exons deletion in SMN1 gene could result in alterations of SMA phenotypes,but it has nothing to do with the severity of SMA.Gene deletion analysis of SMN1 gene can be considered as the preferred fimal diagnosis method for infantile SMA patients.But as for juvenile form,its diagnosis depending on gene deletion analysis of SMN1 gene will be analyzed with precaution.While as for adult form of SMA,the incidence of SMA may be independent with SMN1 gene deletion.Therefore,generally,SMN1 gene assay is not taken as the routine diagnose method for adult form SMA.

Objective To explore the clinical value of genetic diagnosis of SMA,the homozygous deletion of survival motor neuron 1 (SMN1) gene in suspected spinal muscular atrophy (SMA) patients were analyzed in this study.Methods A total of 154 patients suspected with SMA and 20 healthy volunteers were recruited from January 2007 to December 2014 in the Genetic Diagnosis Center of the First People's Hospital of Yunnan Province and the Department of Neurology of the Fourth Affiliated Hospital of Kunming Medical University.Potential deletions in exons 7 and 8 of SMN1 gene were screened by use of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method in both 154 patients suspected with SMA and 20 healthy volunteers.The frequencies of the deletions in exons 7 and 8 of SMN1 were calculated and statistical analysis of different deletion types among 3 SMA groups was performed with SPSS 13.0 software package.Results Among 154 suspected SMA patients,101 cases with homozygous deletions of exon 7 of SMN1 gene were detected,which accounted 65.6％ (101/154) of the suspected SMA patients.Among the 101 SMA patients,97.0％ (98/101) of the patients with both homozygous deletions of exons 7 and 8 for SMN1 gene and 3.0％ (3/101) of the patients with homozygous deletions of only exon 7 for SMN1 gene were detected.The patient with only deletion of exon 8 for SMN1 gene was notdetected.Four cases with negative results were subjected to be followed-up,but they were characteristic of SMA symptom by clinical re-visit.Thus,total 105 patients were confirmed with SMA,among them,68 were type Ⅰ SMA,27 were type Ⅱ SMA,and 10 were type Ⅲl SMA,which accounted for 64.8％ (68/105),25.7％ (27/105) and 9.5％ (10/105) of the SMA patients,respectively.Type Ⅳ SMA was not observed in these patients.No deletion was detected among 20 healthy volunteers.Conclusions PCR-RFLP assay is a noninvasive,simple,high sensitive and specific method for SMA diagnosis,which can be considered as the first-line genetic test for the suspected SMA patients.It will help to improve the accuracy of clinical diagnosis and the detection rate by strengthening the clinical diagnostic criteria and re-evaluating the suspected patients after negative genetic diagnosis.

BACKGROUND: Neural stem cells are always derived from animals, and unsuitable for human transplantation treatment. OBJECTIVE: To explore the in vitro culture methods of human embryonic striatum-derived neural stem cells, and to observe the biological characteristics. METHODS: The human embryonic striatum were separated from the embryo at a gestational age of 8-16 weeks that received induction of labor with water bag, and then the embryonic striatum was in vitro cultured in the serum-free Dulbecco’s modified Eagle’s medium. The cells were passaged after neurospheres formation, and then the cells were induced to differentiation with the Dulbecco’s modified Eagle’s medium/F12 containing 10% fetal bovine serum. RESULTS AND CONCLUSION: The in vitro cultured human embryonic striatum-derived neural stem cells grew rapidly and could express nestin. Colony formation assay showed the cel clone formation rate was 6.0%-7.0%. 5-Bromodeoxyuridine incorporation assay showed the cel proliferation rate was 37.9%. Immunofluorescence staining showed that the cells after induction and differentiation could express Tuj-1, glial fibril ary acidic protein and nestin, but not express myelin basic protein. The results indicate that human embryonic striatum-derived neural stem cells cultured in the serum-free medium can maintain their biological characteristics and have self-renewal capacity, and the cells can differentiate into the neurons and astrocytes induced by the fetal bovine serum.

Objective To investigate the value of nerve conduction studies (NCSs),F wave analysis,somatosensory evoked potential (SEP) and skin sympathetic response (SSR) in the early diagnosis of diabetic peripheral neuropathy (DPN).Methods A total of 110 patients with diabetes mellitus were recruited as the diabetic group and another 50 well-matched healthy volunteers as the normal controls.Sensory and motor NCSs of the median,ulnar,posterior tibial and common peroneal nerves were performed.F waves were recorded from the median and posterior tibial nerves.SEPs elicited by stimulation to nerves of both the upper and lower limbs as well as SSRs were measured,all in both the diabetic group and the normal controls.Results The total rate of nerve conduction abnormality was 74.5％ in the diabetic group,with sensory nerve conduction abnormalities more frequent and more severe among motor nerves in the extremities.The total rate of F wave abnormalities was 57.3％ in the diabetic group.The rate in patients with normal distal motor conduction in their median and posterior tibial nerves was 50.7％.The total SEP abnormality rate was 70.0％ with regard to the proximal peripheral nerve potentials in the diabetic group,but there was no obvious abnormality of the supraclavicular electrical potential in the upper limbs for those with normal sensory nerve conduction in the median nerve.The rate of occurrence of abnormality in the gluteus point potential in the lower limbs of those with normal posterior tibial sensory conduction was 62.5％.The total rate of SSR abnormalities was 80.0％ in the diabetic group but 72％ among those with normal nerve conduction in their extremities.Combining the NCS,SSR,SEP and F wave results,the total abnormality rate was 90.9％ in the diabetic group,which was much higher than with any single test used alone.Conclusion NCS is essential for diagnosing DPN.Early diagnosis of subclinical diabetic neuropathy will be significantly enhanced when nerve conduction,SSRs,SEPs and F waves are tested together.

We transfected human neural stem cells using lentiviral vectors encoding glial cell line derived neurotrophic factor (GDNF) to study its expression level in vitro and to get a stable cell line expressing GDNF. First, GDNF gene was sub-cloned into the lentiviral transfer vectors. Then, the recombinant lentiviral supernatants were packaged by 293T cells through three plasmids transient co-transfection method using standard lipofectamine reagent. The viral titers were tested by the transfection efficiency of 293T cells. At the same time, human neural stem cells (hNSC) were transfected under different multiplicity of infection. GDNF gene expression level and protein secretion level of hNSC were tested by real-time PCR and ELISA methods after transfection. Lentiviral vectors encoding GDNF were constructed. Using lentiviral vectors encoding GDNF we successfully transfected human neural stem cells, and got a stable neural stem cell lines over-expressing GDNF. Furthermore, the results indicated that GDNF expression was influenced by the multiplicity of infection. Human neural stem cells could over-express GDNF through lentivial vectors tranfection. Its gene expression level and protein expression level correlate with the multiplicity of infection.
Embryonic Stem Cells ; metabolism ; Genetic Vectors ; genetics ; Glial Cell Line-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; Nerve Tissue ; cytology ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection