Objective To observe the protective and repairing effects of bone mesenchymal stem cells(BMSCs) transplantation on cerebral infarction in rats and to study the different effect of transplantation at different time points. Methods Middle cerebral artery occlusion(MCAO) models were set up,and the rats were divided into a control group,a group with PBS transplantation and five groups with BMSCs transplantation 3,6,12,24 and 72 h after MCAO,respectively.The volume of infarction area and the neurological severity score(NSS) in all the groups were compared. Results Twenty-eight days after MCAO,the TTC staining indicated that the volume of infarction area in the groups with BMSCs transplantation decreased remarkably compared with the control group and the group with PBS transplantation(P

Objective:To investigate the clinical and molecular genetic features of neonatal congenital Netherton syndrome (NS) caused by mutations in serine protease inhibitor of Kazal type 5 ( SPINK5) gene. Methods:This study retrospectively analyzed the clinical data of an NS neonate admitted to Shanghai Children's Hospital in November 2018. SPINK5 gene was analyzed using high-throughput sequencing and Sanger sequencing. Relevant articles were retrieved from various databases including China National Knowledge Infrastructure, Wanfang and PubMed, and the reported cases who were diagnosed as NS within two months after birth with SPINK5 gene sequencing results were reviewed. Clinical features, gene mutations, treatment and follow-up results of NS were summarized using descriptive statistical analysis. Results:The patient presented with diffuse erythema and desquamation, sparse hair and repeated infections shortly after birth. Laboratory tests revealed elevated IgE (111 IU/ml) and "invagination-like" change in the hair under optical microscope. SPINK5 gene analysis found that there were compound heterozygous mutations of c.2468dup (p.Lys824Glufs*4) and c.377_378del (p.Tys126*) in the child. The pedigree analysis found that the two mutations were respectively inherited from the father and the mother, which supported the diagnosis of NS caused by SPINK5 gene mutation. Though skin rash improved after comprehensive treatments including anti-infection therapy, gamma globulin injection and skincare, the patient suffered from recurrent infection and was discharged from the hospital after giving up treatment and died of infection at two months old. Eleven NS cases were retrieved from literature and altogether 12 cases were analyzed here. The most common clinical manifestations in the 12 patients were early skin diffuse erythema and desquamation (12/12), infection (8/12), dry hair (7/12), hypernatremia dehydration (7/12), high IgE (5/12), growth retardation (4/12), respiratory failure (3/12), atopic constitution (2/12), diarrhea (2/12), dysphagia (1/12), hypothermia (1/12), wheezing (1/12), hypertension (1/12), liver failure (1/12) and metabolic alkalosis (1/12). Conclusions:NS is caused by SPINK5 gene mutation with atypical manifestations in neonates. Neonates with diffuse erythema and desquamation of the skin, repeated infections, dry hair and especially with high blood IgE should be considered the possibility of NS. Genetic testing is conducive to early diagnosis, guiding treatment decisions and providing a basis for genetic counseling.

A 880 bp cDNA localized to the putative NS5 region of HGV genome was expressed in E.coli BL21(DE3). The cDNA fragment was inserted into a plasmid pGEX-5X-1,at the downstream of the DNA sequence encoding Schistosoma japonicum glutathione S- transferase(GST),in the same reading frame with the gene of GST. A 60KD GST-NS53 fusion protein was expressed at 37℃ in a form of inclusion bodies amounting to 30 percent of total host protein whereas at 20℃ mainly in a form of soluble protein . The fusion protein was extracted and purified to homologue. The purified GST-NS53 fusion protein could be specifically recognized with either the sera from the patient infected by HGV or the antisera directed against GST.

lsartan could reduce the release of p-selectin.Conclusions Valsartan could relieve myocardial ischemia reperfusion injury of rat, which may be through reducing p-seleetin of plasma.

A novel method based on the separation and enrichment effect of magnetic beads and the fully complementary hybridization of two DNA strands was developed for highly sensitive detection of bacterial DNA using a surface-enhanced Raman spectroscopy (SERS) with 5,5’-dithio-bis(2-nitrobenzoic acid) (DTNB)-modified gold nanoparticles as reporter probes. Capture probe was immobilized onto the surface of streptavidin-enwrapped magnetic beads ( SA-MB ) through high affinity between biotin and avidin, by which the target bacterial DNA sequences that connected with the reported probe decorated AuNPs with DTNB and SH-DNA ( AuNPs@DTNB@DNA) were captured and loaded onto the magnetic beads by the hybridization reaction with the capture probe. Compared with previous methods, this design shortened the distance between particles by the ways that the magnetic beads tempted to nanoparticles aggregation, and produced the plasma resonance coupling effect, which increased the SERS signal significantly. The results showed that, under the optimized conditions and in the concentration range from 5 pmol/L to 5 nmol/L, the method performed a good linear relationship between Raman intensity and DNA concentration. The limit of detection ( LOD) of bacterial DNA was estimated to be 5 pmol/L. The method is simple and low cost, and can be used in the sensitive and selective detection of bacterial DNA.

Objective To investigate the relationship of epidermal growth factor receptor (EGFR) mutation with clinical features of baselines as well as serum CEA level in patients with recurrent non-small cell lung cancer (NSCLC). Methods A total of 54 patients with first recurrence of advanced lung cancer who had received chemotherapy were included in this study. ADx-ARMS was performed to detect EGFR gene mutations in surgical specimens taken from the primary tumor. Serum CEA level was measured by the electrochemical luminescence method. Results The mutation rate of EGFR was significantly higher in females than in males (χ2= 11.868, P =0.006), with a total mutation rate of 60.8%in 106 patients. The rate was higher in adenocarcinoma than in other histological types(χ2=6.002,P=0.014), and significantly higher in non-smokers than in smokers (χ2= 8.502,P=0.004) and in the patients with serum CEA level over or equal to 5.0 ng/mL than those with CEA level less than 5.0 ng/mL (χ2=22.543,P=0.000). A multivariate analysis revealed that a higher serum CEA level at the time of disease recurrence was associated with EGFR gene mutations (P = 0.002). Conculsions Serum CEA level is closely associated with the presence of EGFR gene mutations in patients with first recurrence of advanced NSCLC. A higher serum CEA level at the time of disease recurrence is independently associated with EGFR gene mutations. CEA level can be used as a potential indicator to determine EGFR mutation.

Objective To screen out the polymerase chain reaction ( PCR) primers for specifically identifying Pheretima aspergillum (E. Perrier) , so as to establish a rapid and accurate method to identify the origin of Pheretima aspergillum. Methods Four kinds of earthworms recorded in pharmacopoeia and six kinds of their adulterants commonly seen in the market were collected. The 12SrRNA sequences related to earthworm were downloaded from GenBank database. PCR amplification for ten kinds of samples was performed using the universal primers, and then the products were sequenced. According to the differences between the above sequences, three pairs of specific primers in the non-conservative district were designed for identification of Pheretima aspergillum. Results High-specificity PCR amplification for Pheretima aspergillum with 12St/12Stf primer occurred when the annealing temperature was increased to 64℃, and only Pheretima aspergillum had single amplification strip, while no strips were found in the other adulterants under the same condition. The achieved specific primers could be well verified in 15 kinds of medicinal materials of earthworm purchased in the market, which had the same morphological features showed by the traditional identifying method. Conclusion With 12St/12Stf primer as a specific marker, the identifica tion of Pheretima aspergillum is rapid and accurate in related species of Pheretima aspergillum, and avoids the effect of some factors such as integrity of experimental materials and drying process.

Objective The establishment of the model of myocardial ischemia-reperfusion injury (MIRI) in SD rats involves lengthy time, severe damage, and death of the rats.We aimed at an efficient and rapid method for establishing an MIRI model in rats to ensure a high success rate of modeling and survival of the animals.Methods Using the trachea intubation-assisted breathing machine, we fully exposed the hearts of 80 adult SD rats (8-10 weeks old and weighing 250-300 mg) by mutilating the 2-3 intercostal muscles.Then, we rapidly positioned and ligated the left anterior descending (LAD) coronary artery for an hour, and established the model of MIRI at 48 hours after releasing the slipknot.We performed electrocardiography (ECG) before, during, and at 1 and 48 hours after operation, ligated the same part again at 48 hours postoperatively, and measured the size of the myocardial ischemia and infarction areas using Evans-TTC double staining.Results The survival rate of the 80 rats was 90% and the success rate of MIRI modeling was 86.25%.After ligation of the LAD coronary artery, ECG showed the ST segment and T wave elevation, followed by gradual decrease of the ST segment and R wave voltage, the myocardium and cardiac apex muscle grey or dark grey below the ligation slipknot.The heart rate myocardial motion amplitude were obviously reduced after ligation.Evans-TTC double staining revealed an evident myocardial infarction area at 48 hours after modeling and the ratio of the ischemia area + infarction area to the total myocardial area of the left ventricle was 0.43 to 0.55 (P>0.05) and that of the infarction area to the ischemia area + infarction area was 0.35 to 0.45 (P>0.05).Conclusion The modified rat model of MIRI can be fast and efficiently established, with a high success rate of modeling and a low mortality of the animals.

OBJECTIVE:To observe the efficacy and safety of spraying mannatide and 5-fluorouracil in anocelia during sur-gery of lung cancer based on the adjuvant chemotherapy. METHODS:Totally 114 patients with lung cancer radical surgery were randomly divided into observation group(59 cases)and control group(55 cases). There were spraying mannatide(40-80 mg)and 5-fluorouracil(500 mg)in the observation group;and nothing drugs were sprayed in control group. The patients higher than phaseⅡ were treated by adjuvant chemotherapy after surgery for continuous 2 cycles,21 d as a cycle. The clinical data was compared, including amount of bleeding and drainage,hospital stay,complications,toxicity,KPS score,body weight changes,immune func-tion,survival rate and recurrence rates between 2 groups. RESULTS:There were no significant differences among the amount of bleeding and drainage,hospital stay and complications in the 2 groups (P>0.05). The nausea,vomiting,leucopenia,the KPS score,improvement of total effective rate,and improvement rate of body mass,immune function,survival rate in 1 and 2 year (s),recurrence rate and time within 2 years in observation group were significantly better than control group,with significant dif-ferences(P<0.01 or P<0.05). CONCLUSIONS:Spraying mannatide and 5-fluorouracil can reduce the toxicity in anocelia during surgery of lung cancer based on the adjuvant chemotherapy,improve the immune function and survival rate,reduce recurrence rate and prolong the recurrence time.