Cholangiocarcinoma (CCA) is a group of solid tumors with high malignant degree and poor prognosis. Surgical resection has still been the main therapy options. Targeting therapy and immunotherapy are the main anti-tumor methods that have been paid more and more attention in recent years, especially immunotherapy. The tumor microenvironment of CCA is complex, which encompasses not only interstitial and endothelial cells, but also a large number of immune cells. In addition, the innate and adaptive immune systems also play a role. This article summarizes the immune-related studies of cholangiocarcinoma and the latest clinical trials of immunotherapy.

OBJECTIVE To investigate bacterial contamination of throat spray in different patient groups before and after using it and explore the satisfactory disinfection methods for decreasing the rate of hospital acquired infection.METHODS The methods were used in accordance with Disinfection Technology Criteria.We compared the germicidal effect of five kinds of methods.RESULTS Bacterial contamination was very serious after using throat spray,many bacteria,even pathogens,as well as the normal flora from the region of nasopharynx had been found there.Bacterial contamination was more serious in ENT department patient than in endoscopy patient after using throat sprays.Burning,alcohol,iodophor and disinfectants containing iodine with alcohol were better than oxidized potential water method on disinfection effect.CONCLUSIONS Much attention should be paid to the contamination of throat spray,selecting the most effective disinfection methods,hospital infections caused by invasive manipulation could be prevented when correct methods are adopted.

OBJECTIVE To study the pathogenicity of the Vibrio fluvialis isolated from the coastal seawater.METHODS Virulence experiment group:22 Kunming mice were divided into four subgroups in random:V.fluvialis was injected into abdominal cavity in the test subgroup.And Staphylococcus aureus and Escherichia coli were injected into the positive control subgroups,separately and aseptic physiological saline was injected into the negative control group.Wound infection group:22 SPF mice were divided into four supgroups in random after their legs were injured:the experimental supgroup(soaked in artificial seawater with V.fluvialis);the positive control groups(with S.aureus and E.coli,separately);the negative control group(soaked in aseptic artificial seawater).The general condition,blood routine,blood culture,organ culture and wound secretion culture of the mice were observed.The pathological analysis of the mice was taken after sacrifice on the 3rd day.RESULTS In virulence experiment group,among all the 7 mice′s blood culture of V.fluvialis supgroup,5 mice were found V.fluvialis positive after 12 h injection,and 2 mice kept on positive until 24 h.In wound infection group,pathological examination showed there were a large number of neutrophils distributed over the striated muscle of the injured sites and cellulitis formed.CONCLUSIONS The V.fluvialis isolated from the sea water has pathogenicity,and can cause wound) infection and septicemia when the concentration reached 106 CFU/ml.

Objective To design and express a novel peptide based on ricin toxin antibody in E. coli, and to evaluate its biological activity. Methods Based on the crystal structure of ricin toxin A chain (RTA) and the RTA-rRNA interact in the complex model, the steric conformation of RTA was theoretical modeled and its functional domain was preliminarily determined. The humanized single-domain RTA antibody was designed rationally by computer-guidod molecular design method. Its coding sequence was ob- tained by overlapping extension PCR, and cloned into the pET-32a vector. The fusion protein was then ex-pressed in E. coli BL21 (DE3), identified by Western blot, and purified with Ni-NTA agarose. The binding and neutralizing activity of this novel peptide for riein was evaluated by competitive ELlSA assay and MTT assay. Results A recombinant human single-domain antibody expressing a polypeptide against RTA in the CDR3 loop was designed. The fusion protein was successfully expressed in E. coll. The purified protein can bind to ricin, and neutralize its activity in SP2/0 viability assay. Conclusion The success of the novel pep-tide based on riein toxin antibody provides a novel method to develop new generation of ricin antagonists.

Objective:To report a Chinese family with a novel ABCD1 gene mutation at c.332T>G (p.V111G) site and discuss its clinical characteristics and molecular mechanism.Methods:The clinical data, laboratory examination, and imaging examination results were analyzed to make the clinical diagnosis of a middle-aged onset patient from the First Affiliated Hospital of Zhengzhou University in May 2017. High-throughput sequencing was used to discover a novel ABCD1 gene mutation. Sanger sequencing was used to find out whether other family members contain the same ABCD1 gene mutation. The pathogenicity of this mutation was explored by protein structure prediction and pathogenicity analysis. Adrenoleukodystrophy protein-green fluorescent protein (ALDP-GFP) and ALDP-GFP (V111G) plasmids were constructed and human embryonic kidney 293 cells were transfected, then immunofluorescence and Western blotting were used to explore the molecular mechanism of this mutation (completed in Henan Provincial People′s Hospital).Results:The proband (a 39-year-old male) was diagnosed as adrenomyeloneuropathy, a subset of X-linked adrenoleukodystrophy, with a novel heterozygous missense mutation in the ABCD1 gene at c.332T>G (p.V111G) site, and his mother and two daughters were all carriers. Protein structure prediction and pathogenicity results suggested that this mutation is pathogenic. Overexpression of ALDP-GFP (V111G) in the human embryonic kidney 293 cells resulted in a significant decrease in the expression levels of ALDP and the abnormal localization from the peroxisomal membrane to the cytoplasm, accompanied by significant down-regulation of LC3-Ⅱ/LC3-Ⅰ and beclin-1.Conclusion:c.332T>G (p.V111G) is a novel pathogenic mutation in the ABCD1 gene, which causes adrenomyeloneuropathy by impairing autophagy.

OBJECTIVE: To observe the curative effects of haemocoagulase on patients with reduced coagulation after the transurethraal plasmakentic vaporization of prostrate(TUPKVP).METHODS: 132 cases were randomly divided into two groups,one treated with haemocoagulase,and the other as control group.Then two groups were compared for the time of operation,quantity of blood transfusion during the operation and the time difference in continuous bladder-washing after the operation.RESULTS: There was an obvious difference(P

Objective To improve the processing protocol of the frozen erythrocyte ( RBC) washing machine of the dialysis type to enhance its efficiency of treating the 2 U frozen RBCs .Methods The frozen RBC washing machine for dialysis was used to deglycerolize the 2 U frozen RBCs after thawing .Measures including adjustment of the Pump speed , washing buffer adding and solution osmolarity were taken to decrease the processing time .Hemoglobin(Hb) content, Hb recovery, residual glycerol, free Hb content, deforming ability, and apoptosis of the deglycerolized RBCs were assessed to evaluate the efficiency of the improved protocol .Results As for the cells processed respectively by the previous and improved protocols, contents of Hb were (35.03 ±4.20) g and (53.82 ±2.08) g, respectively; Hb recoveries were (61.10 ±8.46)% and (80.22 ±3.73)%,respectively.Osmolarities representing residual glycerol were (307.00 ± 14.50)mOsm and (322.00 ±29.00)mOsm.Free Hb values were (0.74 ±0.04) and (0.80 ±0.08)g/L.The processing time was (1.64 ±0.31) and (1.04 ±0.16) h,respectively.There were significant differences in Hb content , Hb recovery, and deforming ability between the cells processed by the two protocols respectively .However , there were no significant changes in glycerol contents , free Hb, and apoptosis extent between the cells processed by the two protocols .Conclusion The innovated protocol can improve the quality of washed frozen RBCs , decrease the processing time and enhance the efficiency of the machine.

Gallbladder cancer is a high malignancy which is predisposed to invade adjacent organs and have lymph node metastasis. Gallbladder cancer is not sensitive to radiotherapy or chemotherapy with the worst prognosis among biliary tract cancers. At present, radical resection is the only possible method to cure gallbladder cancer. However, there are still many controversies about the surgical strategies, the extent of liver resection and lymph node dissection, and the treatment of incidental gallbladder cancer. In addition, under the background of the great success of immunotherapy and targeted therapy in a variety of solid tumors, it is also a question worthy of further considerations that whether the status of surgery in the treatment of advanced gallbladder cancer will be changed in the near future.

Objective To evaluate the application values of diffusion tensor tractography (DTT) combined with neuronavigation in microsurgery of insular gliomas. Methods The clinical data of 27 patients with insular gliomas, admitted to our hospital from March 2013 to October 2017, were analyzed retrospectively. All DTT images were transferred to the neuronavigation system, and the three-dimensional location of tumors and pyramidal tracts were re-constructed. Surgical approaches were designed and excision scopes were defined before the surgery. Techniques on how to distinguish and protect the key blood vessels and pyramidal tracts were discussed. The treatment efficacies were analyzed. Results Total lesion resection was achieved in 22 patients (81.5％), subtotal resection in 4 (14.8％ ), and partial resection in one (3.7％ ). Postoperative pathology indicated 7 were oligodendrogliomas, and 20 were astrocytomas, including WHO grade I in one, grade II in 18, and grade III in one. One patient had transient aphasia (recovery after two weeks), 2 experienced worsened hemiplegia on opposite side of their bodies (normal after one month), and the left 24 patients remained intact function after operation. Conclusions The combination of DTT and neuronavigation is safe and effective in surgical treatment for insular gliomas, which can protect the brain function at greatest degree and maximize lesion resection, and improve the postoperative quality of life.

Objective:To determine the concentration of hydroxychloroquine (HCQ) and its active metabolite deethylhydroxychloroquine (DHCQ) in breast milk of lactating patients with autoimmune disease. To observe the safety of hydroxychloroquine in lactation period, and to explore the factors that may affect HCQ and DHCQ concentration in the milk.Methods:Lactating patients with autoimmune disease who have taken HCQ for at least 6 months were included in our study. A new high performance liquid chromatography (HPLC) method was established to detect HCQ and DHCQ levels in breast milk. Milk samples were collected at different time points: before taking the drug (0 hours), and 2 hours, 4 hours, 6 hours after taking the drug. In addition, the genotype of cytochrome CYP3A4*1G, CYP3A5*3 and CYP2D6*10 which were related to HCQ metabolism were tested by dideoxy chain termination method. Visual acuity, hearing and growth status of the patients' infants were followed up on a regular basis. T-test, one-way ANOVA and Pearson's test were used for data analysis. Results:In 15 patients, the average concentration of HCQ and DHCQ in the milk of patients taking 200 mg/d were (520±261) ng/ml and (177±112) ng/ml, respectively. While the average concentration of HCQ and DHCQ in the milk of patients taking 400 mg/d were (1 036±374) ng/ml and (397±271) ng/ml, respectively. The peak of HCQ level for 11 patients was at 4 hour after taking the drug, while the others' were at 2 hour. The breast-fed infants did not show any abnormal symptoms of hearing, vision and growth. However, cytochrome gene polymorphism did not affect the peak of HCQ and DHCQ.Conclusion:The concentration of HCQ and DHCQ in breast milk is positively correlated to the dosage. The peak level of HCQ milk is 4 hours after taking the drug. The levels of HCQ and DHCQ at 6 hours are similar as those in the whole blood. It is suggested that patients who take HCQ can feed 4 hours after taking the drug to reduce the HCQ and its active metabolites being absorbed by infants. However, the impact of HCQ on infant safety and gene polymorphism of CYP on milk concentration among individuals needs to be further verified in large sample studies and long-term follow-up.