恶性黑色素瘤的发病率和病死率在逐年增长，引发广泛关注。目前恶性黑色素瘤的治疗方式主要为手术、化疗和靶 向治疗，总体疗效极差。然而，PD-1抗体治疗恶性黑色素瘤的成功，使广大研究者把希望聚焦于生物免疫治疗。肿瘤浸润淋巴细 胞（tumor infiltrating lymphocyte，TIL）是从肿瘤组织分离出的淋巴细胞，经体外扩增后得到的能够杀伤肿瘤细胞的细胞群，具有 高杀瘤活性和高靶向性特点。临床试验研究发现，TIL对恶性黑色素瘤治疗效果显著且稳定，但其临床疗效尚未完全阐明。本文 对TIL免疫治疗的发展及其在恶性黑色素瘤治疗中的研究新进展作一综述，以期为恶性黑色素瘤的治疗提供新的策略。

OBJECTIVE:To explore the clinical efficacy and safety of nedaplatin combined with docetaxel in the treatment of advanced cervical cancer. METHODS:A total of 53 patients with advanced cervical cancer selected from our hospital during Apr. 2014-Apr. 2016 were divided into observation group(31 cases)and control group(22 cases)according to chemotherapy plan. Con-trol group was given Docetaxel injection 60 mg/m2,ivgtt,qw. Observation group was additionally given Nedaplatin for injection 35 mg/m2+0.9% Sodium chloride injection diluted into 500 mL,ivgtt (≥60 min),qw. A chemotherapy cycle lased for 21 d,and both groups received 2 cycles of chemotherapy. Clinical efficacies of 2 groups were evaluated 2 weeks after treatment,and the level of PCNA integal was detected before and 2 weeks after treatment. The occurrence of ADR was recorded. RESULTS:The total re-sponse rate of observation group (77.42%) was significantly higher than that of control group (63.64%),with statistical signifi-cance(P<0.05). Before treatment,there was no statistical significance in PCNA integval between 2 groups(P>0.05). After treat-ment,PCNA integval of 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical significance(P<0.05). ADR were concentrated in grade I,and there was no statistical significance in the in-cidence of ADR between 2 groups (P>0.05). CONCLUSIONS:Docetaxel combined with nedaplatin can significantly improve the clinical efficacy of patients with advanced cervical cancer,and does not increase the adverse reactions compared to docetaxel alone.

Objective To observe expression levels of N-methyl-D-aspartate (NMDA) receptor and cholecystokinin (CCK) in the hippocampus and spinal cord in morphine withdrawal or tolerance mice treated by electroacupuncture or catgut embedding and explore the difference between the regulating effects of electroacupuncture and catgut embedding on morphine withdrawal and tolerance.Methods Fifty-six male C57BL/6J mice were randomly allocated to withdrawal control, withdrawal model, withdrawal catgut embedding and withdrawal electroacupuncture groups, and tolerance control, tolerance model, tolerance catgut embedding and tolerance electroacupuncture groups, 7 mice in each group. A model of morphine withdrawal was made by subcutaneous injection of morphine hydrochloride using 7-day increasing addiction method. The withdrawal control group was injected with an equal volume of normal saline at the same time points. In the withdrawal electroacupuncture group, electroacupuncture at bilateral points Shenshu was performed using a Han’s acupoint nerve stimulation device (HANS-200) at 15 min after an injection of morphine hydrochloride. In the withdrawal catgut embedding group, 0.5 cm chromic catgut was embedded in bilateral points Shenshu at 15 min after an injection of morphine hydrochloride. Addiction was promoted by intraperitoneal injection of naloxone 4 mg/kg at 10 o’clock on the seventh day’s morning and Withdrawal reactions were observed in the mice. The score was recorded using the Ryuta Tomoji opioid withdrawal symptoms evaluation scale. NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by enzyme-linked immunosorbent assay (ELISA). A model of morphine tolerance was made by subcutaneous injection of morphine 10 mg/kg. The tolerance control group was injected with tolerance normal saline 10 ml/kg at the same time. In the tolerance catgut embedding group, catgut was embedded in point Shenshu at the first day after model making. In the tolerance electroacupuncture group, point Shenshu was given electroacupuncture at the first day after model making. After seven days of treatment, NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by ELISA.Results There were statistically significant differences in hippocampal NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There was a statistically significant difference in hippocampal NR2B expression between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the withdrawal catgut embedding or withdrawal electroacupuncture group and the withdrawal model group (P<0.05). There were statistically significant differences in spinal cord NR2A, NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There were statistically significant differences in spinal cord NR2A and NR2B expressions between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There were statistically significant differences in hippocampal NR2A, NR2B and CCK expressions between the tolerance model and tolerance control groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the tolerance catgut embedding and tolerance model groups (P<0.05). There was a statistically significant difference in hippocampal NR1 expression between the tolerance electroacupuncture group and the tolerance model or tolerance catgut embedding group (P<0.05). There was a statistically significant difference in spinal cord CCK expression between the tolerance catgut embedding or withdrawal electroacupuncture group and the tolerance model group (P<0.05).Conclusions Both catgut embedding and electroacupuncture at point Shenshu have a reducing effect on morphine tolerance and withdrawal. The therapeutic effect of electroacupuncture is better than that of catgut embedding.

The neuroanatomical basis of meridian effects is one of the focuses of acupuncture mechanism studies. Meridian effects depend mainly on the generation, conduction, integration and output of acupuncture information in the acupoint, spinal cord and brain.The neural tracer technique is a basic method in neurobiology. This article presents recent years' advances in the application of the neural tracer technique to acupuncture field and summarizes the types of neural tracer used in acupuncture studies and regularities in the distribution of neurons and their fibers related to labeled meridians and acupoints in spinal segments and brain regions to provide methodological reference for studies on neurobiological mechanism of acupuncture effects.

Objective:To explore the function of the cya gene and the preliminary mechanism of attenuated strain.Methods:The biological characteristics of cya mutant in acid and alkali resistant,salt resistance,motility,biofilm components,poisonous to the cells of epithelial cell adhesion,invasion were analysis.Results:The mobility capabilities,acid and alkali resistance and salt tolerance of cya mutant were significantly lower than the parent strain;the composition testing revealed that the cya mutant did not produce cellulose,curli and biofilm;at the same time the adhesion and invasion to epithelial cells of cya mutant had a prominent depression,and the toxicity to HeLa cells was weaker than the parent strain.Conclusion:The function of cya gene is closely related to athletic ability, penetration of cell membrane, the formation biofilm and virulence.It will provide a theory reference to the functional research of Salmonella typhimurium cya gene and the mechanism of attenuated strain.This will contribute to the development of oral vaccine using attenuated Salmonella typhimurium as vector.

Objective To research the clinical department distribution and drug resistance of Enterobacteriaceae bacteria to pro‐vide a theoretical basis for clinical rational use of antimicrobial drugs .Methods The Whonet 5 .6 software was adopted to conduct the retrospective analysis on Enterobacteriaceae bacteria isolated in the General Hospital of Ningxia Medical University and the Af‐filiated Cardiovascular Disease Hospital from 2011 to 2013 .Results A total of 7 315 strains of Enterobacteriaceae bacteria were i‐solated ;the top three of bacteria were 2 971 strains (40 .6% ) of Escherichia coli ,2 339 strains (32 .0% ) of K lebsiella pneumoniae and 1 117 strains (15 .3% ) of Enterobacter cloacae ;in the source of specimen ,the respiratory tract specimens had the highest isola‐tion rate (46 .6% ,3 410 strains) ,followed by the pus and secretion specimens (13 .9% ,1 015 strains) ,and the urine specimens (13 .0% ,953 strains) ;the isolated bacterial strains were mainly derived from the pediatric department (17 .5% ,1 282 isolates) ,res‐piration department (7 .1% ,518 strains) and ICU (6 .4% ,468 strains) ;the highest sensitivity of antibacterial drugs were carbapen‐ems ,amikacin and piperacillin/tazobactam also maintained a good antibacterial activity ,the resistance rate was 1 .3% - 7 .6% .Con‐clusion Enterobacteriaceae has a higher isolation rate in the clinical specimens and its resistance rates to antibacterial drugs are generally higher .The surveillance on bacterial drug resistance should be strengthened so as to provide a theoretical basis for the ra‐tional use of antimicrobial drugs and effective control of nosocomial infections .

Objective To study the effect of autologous periosteum on the healing of tendon-bone interface in rabbit rotator cuff tear. Methods Supraspinatus tenotomy was performed on one side shoulder in 60 New Zealand white rabbits to establish the model of rotator cuff tear. The rabbits were randomly divided into two groups:the study group(used autologous periosteum to promote the suture fixation)and control group(simple suture fixation). The rabbits were sacrificed at 4,8 and 12 weeks postoperatively with twenty rabbits sacrificed each time. Tissue samples of the tendon-bone interface was taken for histological examination and biomechanical test was performed to assess the strength of tendon-bone junction. Results At 4 weeks the study group showed extensive inflammatory cell infiltration, and a small amount of chondrocytes and extracellular matrix. At 8 weeks, the study group showed a large amount of immature chondrocytes arranged rather regularly. At 12 weeks,the tendon bone junction appeared similar to normal. In the study group,both the tendon-bone connection and the arrangement of chondrocytes were significantly better than the control group. The result of biomechanical testing showed that the highest tendon load in the study group was significantly higher than the control group (P < 0.05). Conclusions The use of autologous periosteum as a patch to strengthen the repair of the rotator cuff tear can effectively promote the healing of tendon bone interface, shorten the rotator cuff healing time and has good biological properties. This method provides an experimental basis for clinical rotator cuff repair surgery.

Objective: To systematically evaluate the efficacy and safety of PD-1/PD-L1 inhibitor combined with chemotherapy as comparing with chemotherapy alone for the first-line treatment of advanced NSCLC (non-small lung cancer). Methods: RCTs (randomized controlled trials) on PD-1/PD-L1 inhibitor combined with chemotherapy compared with chemotherapy alone for the first-line treatment of advanced NSCLC were searched in the PubMed, Cochrane Library, EMbase, EBSCO, Chinese Biomedical Literature Database (CBM), Chinese Journal Full-text Database (CNKI), and Chinese Scientific Journal Full-text Database (VIP). RevMan 5.2 software was used for the Meta-analysis. Results: Six RCTs with 3 238 advanced NSCLC patients were included in this study. Meta-analysis showed that the combination therapy group was more effective than the chemotherapy alone group in OS (HR=0.86, 95%CI=0.79~ 0.94, P=0.0006) and PFS (HR=0.81, 95%CI=0.78~0.84, P<0.00001). The incidence of adverse reactions, such as thrombocytopenia of grade 1-5, vomiting, diarrhea, hypothyroidism, hyperthyroidism, rash, pneumonitis, colitis, hepatitis, dysgeusia, hepatitis of grade 3-5 and colitis, in combined treatment group were all higher than those in chemotherapy alone group, the differences were statistically significant (P<0.01 or P<0.05). Conclusions: Compared with chemotherapy alone, PD-1/PD-L1 inhibitor combined with chemotherapy can significantly improve the OS and PFS of patients with advanced NSCLC in the first-line treatment, while the overall incidence of adverse reactions is higher than chemotherapy.

BACKGROUND:Collagen is the most abundant extracellular matrix component,which is closely related to the structure and function of the extracellular matrix of skeletal muscle,but the effect and mechanism of recombinant human collagen(rhC)produced by bioengineering technology on the extracellular matrix of skeletal muscle are unclear. OBJECTIVE:To investigate the effect of rhC supplementation on the remodeling of skeletal muscle extracellular matrix after eccentric exercise,thereby revealing the possible mechanism by which rhC improves the injury of skeletal muscle extracellular matrix and promote the recovery after exercise. METHODS:A total of 104 healthy male C57 mice aged 8 weeks old were randomly divided into control group(normal saline),low-dose rhC group(0.2 g/kg),medium-dose rhC group(1.0 g/kg),and high-dose rhC group(2.0 g/kg).Two mice in each group were selected after continuous 7 days of intragastric intervention,and organs were dissected for hematoxylin-eosin staining to determine inflammatory infiltrates.On the 8th day,the remaining mice were subjected to eccentric exercise.The structural changes of the skeletal muscle extracellular matrix were observed under scanning electron microscopy immediately(0),24,48,and 96 hours after eccentric exercise.Meanwhile,grip strength,creatine kinase activity,and protein levels of matrix metalloproteinases 2,9,14 and tissue inhibitor of metalloproteinase-2 in skeletal muscle were detected by western blot assay. RESULTS AND CONCLUSION:Hematoxylin-eosin staining results indicated that short-term rhC supplementation showed no significant effects on the morphology of the heart,liver,spleen and kidney.After one-time eccentric exercise,the recovery rate of grip strength in the medium-and high-dose rhC groups was significantly increased(P＜0.01).The trend of creatine kinase changes was consistent in all groups and there was no significant difference between groups.The recovery process of the extracellular matrix in the low-dose rhC group was faster than that in the control group,and the muscle tract membrane in the medium-and high-dose rhC groups was more complete.The protein level of matrix metalloproteinase 9 in the high-dose rhC group was significantly decreased(P＜0.05).The protein levels of matrix metalloproteinase 14 in the medium-and high-dose rhC groups were significantly decreased(P＜0.05).The protein levels of matrix metalloproteinase 2 in the medium-and high-dose rhC groups were significantly decreased(P＜0.05).Tissue inhibitor of metalloproteinase-2 protein levels in the medium-and high-dose rhC groups were significantly increased(P＜0.05).The ratio of matrix metalloproteinase 2 to tissue inhibitor of metalloproteinase-2 in each rhC group was significantly decreased(P＜0.05).To conclude,pre-supplementation of 1.0 and 2.0 g/kg rhC for 7 days can inhibit extracellular matrix degradation in skeletal muscle after exercise by modulating matrix metalloproteinases and matrix metalloproteinase inhibitors,thereby promoting recovery of skeletal muscle strength in mice.

Objective:To analyze the molecular characteristics of Echovirus 11 (Echo11) strains isolated in Xiangyang, Hubei Province from 2016 to 2017 based on the sequences of VP1 gene.Methods:Rectal and throat swab specimens were collected from children with hand, foot and mouth disease (HFMD) in Xiangyang from 2016 to 2017. Echo11 strains were detected by real-time reverse transcriptase PCR (RT-PCR) and isolated after cultured in human rhabdosarcoma (RD) cells. The VP1 regions of Echo11 strains isolated from RD cells and the whole genomes of three representative Echo11 strains were amplified by conventional RT-PCR and the sequences were analyzed. DNAStar7.0 (MegAlign) and MEGA6.0 (Data) were used to analyze the homology and mutation sites in nucleotide and amino acid sequences. Neighbor-joining method was used to construct phylogenetic trees. Recombination analysis was performed with SimPlot software (BootScanning).Results:A total of 11 Echo11 strains were isolated from 3 494 HFMD cases, accounting for 0.31%. They were highly homologous in the VP1 gene. These strains shared 98.4%-100.0% homology in nucleotide sequences and 98.3%-100.0% homology in amino acid sequences. The homology between the 11 Echo11 strains and the prototype strain (Echo11/Gregory, X80059) was 73.9%-74.8% in nucleotide sequences and 87.7%-88.7% in amino acid sequences. All of the Echo11 strains circulating in Xiangyang were classified into lineage D, having a similarity to the strains circulating in some regions of mainland China since 2013. In multiple regions of the genome, the Echo11 strains isolated in Xiangyang were highly similar to the Henan Echo1 strains in 2010 and the Hubei Echo6 strains in 2015, suggesting there was recombination within the genome of Echo11 strains in Xiangyang.Conclusions:The Echo11 strains circulating in Xiangyang from 2016 to 2017 belonged to lineage D and were recombinant strains.