Objective To explore effect of increasing cerebrospinal fluid (CSF) leptin level by administration from the foramen magnum to the cerebellum medulla oblongata pool in osteoporosis in rabbit model established by ovariectomy (OVX) and methylprednisolones.Methods According to certain inclusion and exclusion criteria,30 adult rabbits which were healthy 5-month-old female New Zealand white rabbits with the same batch and weight were divided into 5 groups (n=6) with random number table,including control group,normal saline group,Low dose group,middle dose group and high dose group.The animal model of osteoporosis (OP) was established 8 weeks after ovariectomy (bilateral ovarian resection) and intramuscular injection of methylprednisolone daily for 8 consecutive weeks in all groups.The control group were only simulated puncture and didn't inject any drugs or normal saline,the normal saline group were intracerebroventricular (ICV) injection with NS and Low,Middle and High dose group injected with low,middle and high dose of leptin respectively at 0,3,7 days after OVX.Before (0 week) and 4,8 weeks after the operation,the measure of dual energy X-ray absorptiometry was done on all the rabbits femur and bone mineral density (BMD) was delivered.Before (0 week) and 4,8 weeks after the first administration,leptin in cerebrospinal fluid (CSF) and serum growth hormone (GH),insulin growth factor 1 (IGF-1) and osteocalcin (OT/BGP) were tested with Enzyme linked immunosorbent assay and serum calcium,phosphorus and alkaline phosphatase (ALP) with automatic biochemical analyzer (ACA).The rabbits were sacrificed and the thalamus and femoral neck were harvested to test the expression of leptin receptor (LEPR) in thalamus and bone morphogenetic protein-2 (BMP-2) and type Ⅰ collagen (Col Ⅰ) at 8 weeks after the first administration.Results Compared with the control group and NS group,the cerebrospinal fluid (CSF) leptin concentration of leptin administration groups kept in a higher level significantly at 4 and 8 weeks after leptin administration,it was positively correlated with the concentration of administration.After administration 4 and 8 weeks,the bone mineral density (BMD) of leptin administration groups were not significantly lower than that of non-administration groups,and the high dose group was unchanged.The serum osteocalcin/bone glad protein (OT/BGP) concentration of leptin administration groups decreased more significantly compared with the non-administration groups at 4 weeks after leptin administration;and it rebounded more significantly than the non-administration groups and low dose group rebounded more significantly than other non-administration groups at 8 weeks.The concentration of serum growth hormone (GH) of 5 groups showed a downward trend at 4 and 8 weeks,the serum GH concentration of leptin administration groups decreased more significantly than the non-administration groups.The concentration of serum insulin-like growth IGF-1 of 5 groups were decreased significantly at 4 weeks,and the serum IGF-1 concentration of control and low dose groups were significantly higher than other 3 groups at 8weeks.Compared with the non-administration groups,the concentration of serum Calcium of leptin administration groups were increased more obvious,and it was positively correlated with the concentration of administration.Serum phosphorus of 5 groups were decreased at 4 weeks and elevated at 8 weeks;there was no difference among the groups.Serum ALP showed a continuous downward trend,and the middle dose group decreased significantly compared with other groups at 4 and 8 weeks.RTPCR results showed that the BMP-2 and Col Ⅰ mRNA expression increased and positively correlated with leptin administration concentration at 8weeks.Conclusion The increase of cerebrospinal fluid leptin level can promote the bone metabolism,reduce bone loss and precaution the occurrence of osteoporosis in rabbit model caused by OVX and methylprednisolone,and it was positively correlated with the concentration of administration.That provides a new method for the prevention and treatment of osteoporosis in rabbit model.

Objective To investigate the enhanced osteogenesis during bone fracture healing after intracerebroven-tricular (ICV) leptin injection, using rabbit model with created segmental bone defect in right tibial. Method Segmental critical-sized defects were created at the right tibial bone of skeletally mature New Zealand white rabbits. In experiment group (leptin group), recombinant rabbit leptin was injected into cerebellomedullary cistern through foramen magnum. While in control group, normal saline was injected in the same way. Bone Mineral Density (BMD) was evaluated by qCT at 1st, 3rd, 7th, 14th and the 21st days. At the 21st days, all rabbits were euthanized to collect the right tibia for histomorphology, to ex-amine the BMP-2 expression in the bone callus by Fluorescence in situ hybridization (FISH). Result In the leptin group, body weight declined more obviously than control group then it start to arise at the 14th day;qCT showed significant higher BMD in leptin group than in control group at the 7th day;FISH showed a higher BMP-2 expression in leptin group than in control group. Conclusion Cerebellomedullary leptin injection through foramen magnum could accelerate limb fracture healing in rabbit model with right tibial bone defection.

Objective To observe whether rabbit blood leptin can leak into the brain after craniocerebral trauma (CCT),and to dynamically detect serum levels of leptin,growth hormone (GH),and insulinlike growth factor-1 (IGF-1),and leptin level in cerebrospinal fluid (CSF) after CCT.Methods The FITC labeled leptin was injected into blood vessels of 15 adult male rabbits after anesthesia.Then 9 rabbits underwent unilateral fluid percussive impact,while 6 rabbits underwent sham operation.Thirty minutes postoperatively,brain tissue was taken to make frozen sections which were used to observe fluorescence labeled leptin range.Thirty three adult male rabbits were randomly divided into serum group,serum-control group,CSF group and CSF-control group.Rabbits in serum group and CSF group received fluid percussive impact,while rabbits in serum-control group and CSF-control group were drilled a 7 mm window in skull.Rabbits in serum group and serum-control group were phlebotomized 2 ml at 1 d,3 d,7 d and 14 d after operation,and rabbits in CSF group and CSF-control group were extracted CSF 0.5 ml at the same time points.Then serum levels of leptin,GH,and IGF-1,and leptin level in CSF were tested by ELISA.Results The fluorescence imaging could be seen in the injured brain tissue of rabbits with CCT,which was more than those in brain tissue of rabbits receiving sham operation.Serum leptin levels of rabbits in serum group at each time point were higher than those in serum-control group.Serum levels of GH and IGF-1 and leptin level in CSF were higher in rabbits with CCT than those in rabbits without CCT at 1 d,3 d and 7 d after operation.Conclusion The blood leptin can leak into the brain after CCT,which can cause increase of blood GH and IGF-1.And the latter may be the endocrine factors promoting fracture healing after CCT.

OBJECTIVE:To systematically review therapeutic efficacy of Haikun shenxi capsule in the treatment of chronic renal failure (CRF).METHODS:Retrieved from Central,PubMed,EMBase,CJFD,CBM,VIP and Wanfang database,randomized clinical trials (RCTs) about Haikun shenxi capsule combined with routine treatment (trial group)vs.single routine treatment (control group) in the treatment of CRF were collected.Meta-analysis was performed by Rev Man 5.3 software after screening literature,extracting data and evaluating quality by using risk bias evaluation of Cochrane collaboration network.RESULTS:A total of 10 RCTs were included eventually,involving 704 patients.The results of analysis showed that compared to general therapy,Haikun shenxi capsule could improve total effective rate [RR =4.42,95 ％ CI (2.70,7.22),P＜ 0.001],and could reduce SCr[MD =-140.37,95 ％ CI (-191.72,-89.03),P＜ 0.001],BUN[MD =-5.49,95 ％ CI (-8.36,-2.63),P＜ 0.001] and 24 h-Upro [MD =-0.43,95 ％ CI (-0.62,-0.23),P＜ 0.001],with statistical significance.CONCLUSIONS:The clinical efficacy of Haikun shenxi capsule in the treatment of CRF is good and significantly improve related indexes of renal function.

OBJECTIVE: To evaluate the effect of image guided navigation system (IGNS) in complex endoscopic endonasal surgery. METHOD: Three hundred and five patients were undergone endoscopic surgery using IGNS. The detailed information and data including the IGNS accuracy, the operation procedure time, the operator's satisfaction degree and self-confidence, and the perioperative complication were analysed and summarized to evaluate the effect of IGNS. RESULT: The operation procedures were successfully performed in all the 305 patients without perioperative complication. In 98.7% of the total operation IGNS provided an accurate anatomical localization less than 2 mm localization error (0.5-2.0 mm,mean 1.2 mm). The application of IGNS reinforced the operators self-confidence, increased the patients' safety with the almost same time as that of routine endoscopic surgery (ESS). CONCLUSION IGNS could elevate the effectivity and security of ESS, indicating it's a valuable tool in deal with complex endoscopic endonasal surgery.
Adolescent ; Adult ; Child ; Endoscopy ; methods ; Female ; Humans ; Male ; Middle Aged ; Nose ; surgery ; Software ; Surgery, Computer-Assisted ; Tomography, X-Ray Computed ; Young Adult

BACKGROUND:The studies have shown that the mesenchymal stem cel s derived from bone marrow and umbilical cord can be continuously cultured in vitro, and maintain the characteristics of stem cel s. The mesenchymal stem cel s can differentiate into hepatocyte-like cel s after“cocktail”induction by various cytokines. OBJECTIVE:To further identify whether umbilical cord-derived mesenchymal stem cel s in vitro co-cultured with normal hepatocytes can differentiate into hepatocyte-like cel s, and to investigate the differentiation method. METHODS:Mesenchymal stem cel s were isolated from human umbilical cord with adherent method, and the surface markers of umbilical cord-derived mesenchymal stem cel s were detected with flow cytometry. The umbilical cord-derived mesenchymal stem cel s were co-cultured with liver LO2 cel s without adding exogenous inducers. The expressions of alpha-fetoprotein, albumin and human cytokeratin 19 mRNA of hepatocyte specific markers were detected with reverse transcription PCR at 7, 14 and 21 days after culture, and periodic acid-Schiff staining was used to identify the functions. RESULTS AND CONCLUSION:Mesenchymal stem cel s could isolated from human umbilical cord successful y, showing fibroblastic morphology and adherent cel characterization. Among these cel s, 96.02%cel s were CD29 positive cel s and 96.6%cel s were CD105 positive cel s. The percentage of CD34 negative cel s was 99.65%. The percentage of CD105+CD29+double positive cel s was 94.84%. The mRNA of alpha-fetoprotein was found on the 7th day after co-cultured with LO2 cel s, and the mRNA of albumin and human cytokeratin 19 were found on the 14th day. After co-cultured for 21 days, the alpha-fetoprotein mRNA could not be observed in the co-culture group. The expressions of albumin and human cytokeratin 19 were increased at 14 days. After co-cultured for 21 days, the glycogen staining was positive. Umbilical cord-derived mesenchymal stem cel s can differentiate into hepatocyte-like cel s after co-cultured with normal hepatocytes.

Aim To study the apoptosis effect of cucurbitacin Ⅱa on non-small cell lung cancer cell lines NCI-H460 and A549 and its underlying mechanism.Methods Cell viability was assessed by CCK-8 assay.The apoptosis effect and cell cycle arrest were detected by Flow cytometry.Western blot was employed to detect the related protein.Results The proliferation of lung cancer cell lines NCI-H460 and A549 was inhibited by CuⅡa, which showed cytotoxic activity with IC50 values of 224.9 nmol·L-1 and 108.3 nmol·L-1 against NCI-H460 and A549 respectively.CuⅡa induced the cells apoptosis and cell cycle arrest at G2/M phase.The results of Western blot showed CuⅡa inhibited the phosphorylation of STAT3 and Cofilin in a dose-dependent manner.Further, CuⅡa inhibited the phosphorylation of Aurora A, in line with the important characteristics of anti-tumor effect of Aurora A kinase inhibitor with blocking cells in the G2/M phase.Conclusion CuⅡa has obvious anti-tumor effect against non-small cell lung cancer, which suggests its value as a lead compound for lung cell carcinoma.

Objective:To study the expression of Toll like receptor 3 (TLR3) in human adenocarcinoma of the lung cells induced by respiratory syncytial virus (RSV) and its significance in the diagnosis of pneumonia in children.Methods:A549 cells were divided into RSV infection group [added 1 μg/ml Lipopolysaccharide (TLR3 agonist) transfected RSV virus after 150 μl intervention], Lipopolysaccharide stimulation group (added 1 μg/ml Lipopolysaccharide 150 μl intervention) and normal control group (normal culture). The mRNA expressions of tumor necrosis factor-α, interleukin 8, TLR3 protein and TLR3 in A549 Cells of different groups were compared. We prospectively selected 80 children with RSV infectious pneumonia admitted to Baoding Second Central Hospital from August 2019 to October 2021 as the RSV pneumonia group, and sixty children with common pneumonia were taken as the common pneumonia group, and 60 healthy children in our hospital were taken as the control group. The mRNA expression of serum TLR3 in different groups was compared, and the diagnostic efficacy of serum TLR3 in RSV pneumonia was evaluated by receiver operating characteristic.Results:There was a statistically significant difference in the expression of TLR3 protein among different groups of A549 cells ( P<0.001). The expression differences of TLR3 mRNA in different groups of A549 cells at different time points were statistically significant(all P<0.001). There was significant difference in the expression of tumor necrosis factor-α and interleukin 8 of A549 cells at different time points in different groups (all P<0.05). There was a statistically significant difference in the expression of serum TLR3 mRNA among the three groups of subjects ( F=155.237, P<0.001). The critical value for TLR3 gene diagnosis was 66.87, with corresponding sensitivity of 73.75%, specificity of 70.83%, and the area under curve (AUC) of 0.803(95% CI: 0.753-0.855). Conclusions:Respiratory syncytial virus induces human lung cancer cells and promotes disease progression through TLR3 expression; Serum TLR3 can be used for the diagnosis of RSV pneumonia.

BACKGROUNDMany researches demonstrate that the secondary brain injury which is caused by autoimmune attack toward brain antigens plays an important role in surgical brain injury (SBI). Although traditional immunosuppression can reduce autoimmune attack, it will lower the body immunity. Immune tolerance, by contrast, not only does not lower the body immunity, but also could lighten autoimmunity. This study used thymus tolerance to develop an immune system that is tolerant to autologous cerebrospinal fluid (CSF) and autologous brain tissue so that autoimmune injury can be suppressed following the disruption of the blood-brain barrier, thereby reducing brain damage.

METHODSEighty experimental rabbits were divided into five groups by random number table method: 16 in SBI group (group A), 16 in SBI+CSF drainage group (group B), 16 in SBI+CSF drainage+PBS injection group (group C), 16 in SBI+CSF drainage+CSF intrathymic injection group (group D), and 16 in SBI+brain homogenate intrathymic injection group (group E). Rabbits' CSF was drained in group B; was drained and injected PBS into thymus in group C; was drained and injected CSF into thymus in group D; and was injected brain homogenate in group E. Half of the rabbits in each group were phlebotomized on 1st, 3rd, 7th, and 14th days to observe the changes in IL-l, TGF-β by ELISA test, and CD4CD25 regulatory T cells ratio by flow cytometry, and in other animals brain tissues were taken on 7th day for exploring FasL expression by RT-PCR. The least significant difference (LSD) test was used to make paired comparisons; P < 0.05 was considered statistically significant.

RESULTSThe levels of FasL, TGF-β, and the ratios of CD4CD25 regulatory T cells in groups D and E were apparently higher than those in other three groups (P < 0.05). Likewise, the levels of IL-1 in these two groups were lower than the other three groups (P < 0.05). Moreover, the ratios of CD4CD25 regulatory T cells and the levels of TGF-β in groups B and C were higher than those in group A, but the level of IL-1 was lower than that in group A (P < 0.05). There was no significant difference between groups B and C, and groups D and E.

CONCLUSIONThymic injection of CSF and brain homogenate may be able to reduce inflammation after SBI, so thymus immune tolerance may be a useful therapy to treat SBI.

Animals ; Autoantigens ; administration & dosage ; Brain ; surgery ; Brain Injuries ; etiology ; therapy ; Immune Tolerance ; physiology ; Rabbits ; Thymus Gland ; immunology

Objective:To summarize the clinical characteristics of patients with coronavirus disease 2019 (COVID-19) infected with Delta variant, so as to provide further references for clinical diagnosis and treatment.Methods:A real-world study was conducted to analyze the characteristics of 166 COVID-19 patients infected with Delta variant at Guangzhou Eighth People’s Hospital, Guangzhou Medical University.Results:The study enrolled 5 asymptomatic cases, 123 non-severe cases (mild and moderate type), and 38 severe cases (severe and critical type). Among these patients, 69 (41.6%) were male and 97 (58.4%) were female, with a mean age of 47.0±23.5 years. Thirty-nine cases (23.5%) had received 1 or 2 doses of inactivated vaccine. The incidence of severe COVID-19 cases was 7.7% in 2-doses vaccinated patients, which was lower than that of 11.5% in 1-dose and 26.8% in unvaccinated patients. The proportion of severe cases in 2 dose-vaccinated patients was 7.7%, which was lower than that of 11.5% in 1-dose vaccinated patients and 26.8% in unvaccinated patients, but the difference was not significant ( P>0.05). The most common clinical symptom was fever (134 cases, 83.2%), and 39.1% of cases presented with high-grade fever (≥39 °C); other symptoms were cough, sputum, fatigue, and xerostomia. The proportion of fever in severe cases was significantly higher than that of non-severe cases (97.4% vs. 76.4%, P<0.01). Similarly, the proportion of severe cases with high peak temperature (≥39 ℃) () was also higher than that of non-severe cases (65.8% vs. 30.9%, P<0.01). The median minimal Cycle threshold (Ct) values of viral nucleic acid N gene and ORFlab gene were 20.3 and 21.5, respectively, and the minimum Ct values were 11.9 and 13.5, respectively. Within 48 h of admission, 9.0% of cases presented with decreased white blood cell counts, and 52.4% with decreased lymphocyte counts. The proportions of increased C-reactive protein, serum amyloid A, interleukin 6, and interleukin 10 were 32.5%, 57.4%, 65.3%, and 35.7%, respectively. The proportions of elevated C-reactive protein, serum amyloid A and interleukin-6 in severe cases were significantly higher than those in non-severe cases ( P<0.01). Logistic regression analysis showed that older age and higher peak temperature were associated with a higher likelihood of severe cases ( OR>3, 95% CI: 2-7, P<0.01). In terms of treatment, traditional Chinese medicine (TCM) was used in 97.6% of non-severe cases and 100% in severe cases. Other treatments included respiratory and nutritional support, immunotherapy (such as neutralizing antibodies and plasma of recovered patients). The median times from admission to progression to severe cases, of fever clearance, and of nucleic acid conversion were 5 days, 6 days and 19 days, respectively. No deaths were reported within 28 days. Conclusions:The symptoms of Delta variant infection in Guangzhou are characterized by a high proportion of fever, high peak temperature, long duration of fever, high viral load, a long time to nucleic acid conversion, and a high incidence of severe cases. The severe cases exhibit a higher percentage of elderly patients, a longer duration of fever and have a higher fever rate and a higher hyperthermia rate than non-severe cases. Age and hyperthermia are independent risk factors for progression to severe disease. The combination of TCM and Western medicine can control the progression of the disease effectively.