BACKGROUND:Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital,Beijing University of Chinese Medicine,with clear clinical efficacy,but the specific mechanism needs to be elucidated. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group(n=12)and a model group(n=52).An animal model was established by the Rice-Vannucci method.After successful modeling,52 model rats were randomly divided into control model group(n=12),minocycline group,and the low-,medium-,and high-dose groups of the Shujin Jiannao Prescription(n=10 per group).Rats in the minocycline group were given 40 mg/kg·d minocycline by gavage;rats in the low-,medium,and high-dose groups were given 4,8,and 16 g/kg·d Shujin Jiannao Prescription granules by gavage,respectively;and rats in the normal group and control model group were given an equal dose of normal saline by gavage.Medication in each group was given once a day for 1 week.The rats in each group were evaluated behaviorally using suspension test,abnormal involuntary movement score,and Bederson score.The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining.The levels of tumor necrosis factor α,interleukin 1β,and interleukin 10 in the cerebral cortex were determined using ELISA.The positive expressions of Janus kinase 2(JAK2),phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)in the cerebral cortex were detected using immunohistochemistry.The protein expression levels of JAK2,p-JAK2,and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION:Compared with the normal group,the suspension test score and involuntary movement score were decreased in the control model group(P＜0.01 or P＜0.05).The pathological results showed structural disruption of nerve cells,formation of large numbers of vacuoles,cell swelling,and increased intercellular space in the control model group.In addition,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased(P＜0.01),the expression of interleukin 10 was decreased(P＜0.05),and the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly increased(P＜0.01)in the control model group compared with the normal group.Compared with the model group,minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats(P＜0.01 or P＜0.05)and ischemic-hypoxic pathological changes were attenuated,with only a small amount of necrotic nerve cells and a few vacuoles,and reduced intercellular space.Moreover,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group(P＜0.05),while the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly decreased(P＜0.01).The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group.To conclude,Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury.The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway.

Heart rate recovery,a widely used indicator in clinical evaluation of the autonomic nervous system,refers to the difference between maximum heart rate during exercise and the heart rate after 1 or a few minutes of cessation.It possesses advantages of being noninvasive and repeatable.Currently,heart rate recovery is extensively applied in cardiovascular diseases for evaluating disease severity,prognosis,risk of cardiovascular events,and mortality.This article primarily discusses the application value and current research progress of heart rate recovery in cardiovascular diseases.

The stem and branch extract of Tripterygium wilfordii (Celastraceae) afforded seven new dihydroagarofuran sesquiterpene polyesters [tripterysines A-G (1-7)] and eight known ones (8-15). The chemical structures of these new compounds were established based on combinational analysis of HR-ESI-MS and NMR techniques. The absolute configurations of tripterysines A-C (1-3) and E-G (5-7) were determined by X-ray crystallographic analysis and circular dichroism spectra. All the compounds were screened for their inhibitory effect on inflammation through determining their inhibitory effect on nitric oxide production in LPS-induced RAW 264.7 cells and the secretion of inflammatory cytokines TNF-α and IL-6 in LPS-induced BV2 macrophages. Compound 9 exhibited significant inhibitory activity on NO production with an IC₅₀ value of 8.77 μmol·L^-1. Moreover, compound 7 showed the strongest inhibitory effect with the secretion of IL-6 at 27.36%.
Tripterygium/chemistry* ; Esters/pharmacology* ; Interleukin-6 ; Lipopolysaccharides/pharmacology* ; Plant Leaves/chemistry* ; Anti-Inflammatory Agents/chemistry* ; Nitric Oxide/analysis* ; Sesquiterpenes/chemistry* ; Molecular Structure

Objective:To evaluate bone formation in human extraction sockets with absorbed surrounding walls augmented with Bio-Oss(R) and Bio-Gide(R) after a 6-month healing period by histologic and histomorphometric analyses.Methods:Six fresh molar tooth extraction sockets in 6 patients who required periodontally compromised moral tooth extraction were included in this study.The six fresh extraction sockets were grafted with Bio-Oss(R) particle covered with Bio-Gide(R).The 2.8 mm × 6.0 mm cylindric bone specimens were taken from the graft sites with aid of stent 6 months after the surgery.Histologic and histomorphometric analyses were performed.Results:The histological results showed Bio-0ss(R) particles were easily distinguished from the newly formed bone,small amounts of new bone were formed among the Bio-Oss(R) particles,large amounts of connective tissue were found.Intimate contact between the newly formed bone and the small part of Bio-Oss(R) particles was present.All the biopsy cylinders measurement demonstrated a high inter-individual variability in the percentage of the bone,connective tissues and BioOss(R) particles.The new bone occupied 11.54％ (0-28.40％) of the total area;the connective tissues were 53.42％ (34.08％-74.59％) and the Bio-Oss(R) particles were 35.04％ (13.92％-50.87％).The percentage of the particles,which were in contact with bone tissues,amounted to 20.13％ (0-48.50％).Conclusion:Sites grafted with Bio-Oss(R) particles covered with Bio-Gide(R) were comprised of connective tissues and small amounts of newly formed bone surrounding the graft particles.

Objective To investigate the effects of human IL-18 gene combined with diterpenoid alkaloids in inhibiting the proliferation and inducing the apoptosis of tongue squamous carcinoma cells Tscca.Methods We constructed recombinant plasmid pEGFPN3-IL-18 and tranfected it into tongue squamous carcinoma cells Tscca.The transduction efficiency of the target cells was detected by fluorescent microscopy,cytotoxic effect of IL-18 gene with diterpenoid alkaloids on Tscca was detected by MTT assay,and apoptosis was detected by flow cytometry. Western blot was employed to examine the expression level of cellular signal-regulated kinase Akt/p-Akt.Results The tongue squamous cells Tscca which transfected pEGFPN3-IL-18 had significantly increased apoptosis compared with non-transfected cells (P<0 .05 ).Tongue carcinoma squamous cells cultured with diterpenoid alkaloids at the concentrations of 0 .2 ,0 .4 and 0 .6 mg/mL had significantly increased apoptosis in a dose-dependent manner (P<0.05).Human IL-18 gene combined with diterpenoid alkaloids for 48 hours inhibited significantly Tscca in a concentration-dependent manner compared with diterpenoid alkaloids alone (P<0 .05 ).The two in combination could also decrease the protein level of p-Akt dose-dependently.Conclusion The combination of pEGFPN3-IL-18 and diterpenoid alkaloids has a synergistic effect in inhibiting the growth of tongue squamous carcinoma cells Tscca.

Objective:To investigate the changes of the vertical height and width of the alveolar bone six months after the alveolar ridge preservation in periodontal compromised molar sites of severe alveolar bone defects with clinical direct measurement,parallel periapical radiographs,and cone-beam computed tomography (CBCT),and to analyze the effect of the three different methods of measurement.Methods:In this study,20 subjects requiring tooth extraction on account of periodontal disease with a total of 23 ex-tracted molars were enrolled.Extractions were performed atraumatically and patients were received alveo-lar ridge preservation procedure with Bio-Ossand Bio-Gide.Clinical direct measurements were taken after tooth extraction and during the implant surgery 6 months later,CBCT scans and parallel periapical radiographs were taken immediately after ridge preservation and 6 months later.The changes of alveolar ridge width and vertical height after six months were measured and analyzed through the above-mentioned three methods and the similarities and differences of the measured effect were compared.Results:There were no significant difference of alveolar vertical height in the center of the extraction sites,the center of distal aspect,and distobuccal aspect between the clinical direct measurements and the CBCT measure-ments (P>0.05),alveolar vertical height in other points and alveolar width measurements were statical-ly significant (P<0.05).After 6 months,1 0 sites of 1 0 subjects were received a flap and re-entered to perform dental implants surgery.The vertical height in the center of alveolar increased significantly and the changes of alveolar vertical height of clinical direct and CBCT measurement were (6.1 5 ±1 .73)mm and (6.59 ±2.53)mm,respectively.The measurements of the width of the alveolar bone were (8.45 ± 1 .1 8)mm and (8.52 ±1 .27)mm,respectively.The measurements of the two methods were not statisti-
cally significant (P>0.05).The change of the alveolar height in the center of the extraction socket after six months measured by parallel periapical was (5.84 ±4.28)mm,which was closed to the clinical di-rect measurement and the CBCT measurement.Conclusion:Clinical direct measurement and CBCT measurement were largely consistent in the evaluation of the alveolar bone height and width after the alveolar ridge preservation using deproteinized boving bone mineral (DBBM,Bio-Oss)and bioabsor-bable collagen membrane (Bio-Gide)in periodontal compromised molar sites of severe bone defects.

Objective:To discuss the risk factors in lung cancer patients with chemotherapy-induced severe neutropenia to provide reference for clinical drug use. Methods:A retrospective analysis was performed for the patients with lung cancer,and the risk factors of severe neutropenia were statistically analyzed and found out. Results:The results of single factor experiments showed that the incidence of severe neutropenia was related with radiotherapy history,cycles of chemotherapy and the use time of granulocyte colony factor. Based on a binary logistic regression analysis,the history of radiotherapy and the use of granulocyte colony factor were the significant risk factors of severe neutropenia in the lung cancer patients. Conclusion:For the patients with radiotherapy history,it is better to choose chemotherapy drugs with lower toxicity,decrease drug dosage or preventively use granulocyte colony factor. The rational use of rhG-CSF can alleviate chemotherapy-induced severe neutropenia.

Objective To analyze risk factors and complication characteristics of healthcare-associated infection (HAI)in patients with lung cancer,and provide evidence for the formulation of HAI management strategy. Methods HAI-related articles were retrieved from China Biology Medicine (CBM),China National Knowledge Infrastructure (CNKI),Wanfang database,Vip database,PubMed,and Embase,all data were conducted Meta-analysis.Results A total of 19 articles involving 8 069 hospitalized patients with lung cancer (1 280 had HAI)were included.Meta-analysis on combined values of medical factors for HAI were as follows:OR(95%CI )of anti-tumor therapy(radiotherapy and chemotherapy),number of chemotherapy (≥ 2 times ),antimicrobial prophylaxis, immunosuppressant therapy,and invasive operation were 3.13 (1 .82,5.39),9.20 (3.04,27.87),3.23 (1 .77, 5.91),2.00(1 .56,2.57),and 2.28(1 .81 ,2.88),respectively;Meta-analysis on combined values of complication factors for HAI were as follows:OR (95% CI )of pulmonary diseases,chronic obstructive pulmonary disease (COPD),diabetes,renal dysfunction,malnutrition,hypoalbuminemia,neutropenia,and leukopenia were 2.65 (1 .74,4.02),2.40 (1 .76,3.27),2.25 (1 .85,2.73 ),2.56 (1 .18,5.52),5.51 (1 .70,17.89),2.05 (1 .56, 2.70),3.38(1 .40,8.18),and 2.10 (1 .22,3.62),respectively.Conclusion HAI-related factors of medical treat-ment and complications in patients with lung cancer are diversity,risk factors for HAI in patients with lung cancer are anti-tumor therapy,immunosuppressant therapy,antimicrobial prophylaxis,invasive operation,pulmonary dis-eases,COPD,diabetes,renal dysfunction,malnutrition,hypoalbuminemia,neutropenia,and leucopenia.

Objective To evaluate in vitro effects of specific small interfering RNA (siRNA)-silencing of the casitas B-lineage lymphoma b (Cbl-b)gene on immunocompetence of primary murine lymphocytes. Methods Spleens were resected from C57BL/6 mice, and splenic lymphocytes were sterily isolated and cultured in vitro. These lymphocytes were divided into 3 groups: silence group transfected with a Cbl-b-specific siRNA using the EntransterTM-R 4000 reagent, negative control group transfected with a negative control siRNA using the EntransterTM-R4000 reagent, blank control group receiving no treatment. After additional culture for 72 hours, ELISA was performed to measure levels of interferon γ(IFN-γ)and tumor necrosis factor α (TNF-α)in culture supernatants of lymphocytes. In addition, the Cbl-b gene-silenced lymphocytes were co-cultured with B16F10 melanoma cells to evaluate their immunocytotoxic effects on melanoma cells. Results Splenic lymphocytes were successfully isolated from C57BL/6 mice and cultured in vitro, and the Cbl-b-specific siRNA was also successfully transfected into the primary murine lymphocytes and effectively down-regulated the expression of Cbl-b gene in them. Compared with the negative control group and blank control group, the silence group showed significantly increased supernatant levels of IFN-γ and TNF-α(all P < 0.05). The immunocytotoxic effect of lymphocytes on melanoma cells was significantly stronger in the silence group than in the negative control group. Conclusion Cbl-b gene silencing can promote secretion of IFN-γ and TNF-α by murine lymphocytes, and enhance their immunocytotoxic effects on B16F10 melanoma cells in vitro.

Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06％ ± 6.37％ vs.100.00％ ± 10.42％ and 108.70％ ± 9.90％ at 24 hours,42.93％ ± 5.93％ vs.100.00％ ± 6.24％ and 168.00％ ± 2.98％at 48 hours,all P ＜ 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P ＜ 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P＜ 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P＜ 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P ＜ 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P ＜ 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30％ vs.0.421％ and 2.22％ at 24 hours,4.06％ vs.0.577％ and 0.691％ at 48 hours,all P＜ 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.