This paper deals with the ultrastructure of the body wall of adult Pagumogonimus skrjabini by transmission electron microscopy. Infected crabs were collected from Siyen, Hubei Provine, and adult worms were obtained from the lungs of experimentally infected dogs 90 days post-infection.The normal structure of body wall of the P. skrjabini is composed of tegument, tegument cell, musele, muscle cell and protoplasmic tubules, all of which form together syncytium.The tegument contains external plasma membrane, tegument matrix and basal plasma membrane. The cell coat in fine granules is distributed over the whole external plasma membrane surface. The tegument matrix contains various secretory bodies, vesicles and mitochondria. The tegument cell is irregular in shape. Golgi complex, ribosome, autoly-sosome are seen in the cytoplasm. There are two layers of muscle, the external circular muscle and the inner longitudinal muscle layers. The nucleus of immature muscle cell has many heterochromatins, while the nucleus of mature muscle cell is large and round in shaps. Mitochondria and glyco-gen granules are transmitted from muscle cell proper to the muscle by protoplasmic tubules (Figs. 1-7).

Objective To analyze the characteristics of blood loss for laboratory test of critically ill premature infants,and to seek feasible measures to reduce the blood loss.Methods Two hundred and forty-six cases of critically ill premature infants admitted to the neonatal intensive care unit from April 2012 to April 2013 were analyzed the blood loss for test during the hospitalization and the clinical features of blood loss with different gestational age,different weight within the first four weeks after admission.Then the application of blood loss according to test category was described.The blood volume demanded in theory was determined by the formula B =5 (∑ s + 0.1),then calculated the phlebotomy overdraw on the basis of the practical blood loss and analyzed the characteristics of overdraw per patients per day in first two weeks after admission.Results Among 246 patients,The median blood loss figure was 25.57 ml for each infants with the range between 7.10 ml ～ 119.20 ml,and the blood loss concentrated in first four weeks,which showed a decreasing trend with time.There was a statistically significant difference(P ＜0.05)that the smaller gestational age,the lower birth weight,the more daily blood loss per patient per day in first two weeks,but no significant differences(P ＞ 0.05) between the third and fourth week.The largest proportion of the blood samples was used for clinical chemical tests(31.49％),followed by blood gas analysis (19.03％),immunoassays (12.69％),blood cultures (12.63％),hematology (12.28％).The practical blood loss was about twice times of blood volume demanded for tests in theory,which the median was 7.8 times to the latter(25.57 ml vs 3.26 ml).It showed statistically significant difference(P ＜0.05) between blood overdraw per patients per day in the first week and the second week.Compared with different gestational age and birth weight,the difference of overdraw was also statistically significant(P ＜0.05).According to test category,blood culture was the most significant samples of phlebotomy overdraw,followed by biochemical,other,blood gas analysis,the percentage was 76％,64％,45％ and 41％ respectively.Conclusion The blood loss for laboratory test and the phenomenon of blood waste is serious in critically ill preterm infants.The smaller the gestational age is,the lower the weight is,the amount of blood loss and phlebotomy overdraw are more significantly.Biochemical and blood gas analysis are the main items of blood loss.

Objective To investigate the surgical procedure of restoring stenosed hilar bile duct by gallbladder lamella with cystic artery.Methods Based on extensive incising and hilar cholangioplasty,stenosed hilar bile duct was restored by gallbladder lamella with cystic artery.Results Among all the 12 pa tients,including 10 intrahepatic bile duct calculi patients,1 right hepatic duct injury and 1 left intrahepatic chaolangiocarcinoma,no cystic fistula,reflexed cholangitis and recurrent stone were noticed by post operative follow-up.Conclusion This procedure is a ideal method by remaining the normal function of Oddis sphincter while removing stenosis.

AIM:To elucidate the molecular mechanisms of lipoic acid (LA) on redox regulation and digestive function in intestine of C57BL/6 mice fed high fat diet (HFD).METHODS:C57BL/6 mice were randomly assigned to one of three groups (n=8). The control group consumed an ordinary diet. The other two experimental groups were fed with a high fat diet,high fat plus 0.1% LA. After 6 weeks,the activities of digestive enzymes were examined. In order to evaluate the antioxidant status of the mice,superoxide dismutase (SOD),malondialdehyde (MDA),total antioxidant capacity (TAC) and reactive oxygen species (ROS) in intestinal homogenate were measured. To investigate the molecular mechanisms underlying the effects of LA,the gene expression profiles in intestine were examined using the GeneChip microarray system.RESULTS:A depressed antioxidant defense system,accompanied by digestive and absorptive function impairment,was observed in HFD-fed mice. These changes were partially restored in the LA-treated group. DNA microarray analysis of intestine showed that LA ingestion up-regulated the expression of genes related to free-radical scavenger enzymes,digestive enzymes and transporters.CONCLUSION:Treatment with LA improves redox homeostasis and the function of intestine in mice fed HFD. The mechanism may involve preventing oxidative stress by scavenging ROS directly and increasing those of free-radical scavenger enzymes gene expression indirectly.

Objective To explore the influence of high fat diet on the intestinal gene expression profile in C57BL/6 mice. Method C57BL/6 mice were randomly assigned to two groups (n=8). The control group consumed an ordinary diet. The experimental group was fed with a high fat diet. All mice were sacrificed at the end of 6 w and the intestinal gene expressions were detected by oligonucleotide microarray analysis with Affymetrix GeneChip Mouse U430A consisting of 13 097 genes. Results Among the 13 097 genes obtained from gene expression profile analysis, there were 88 and 179 genes up -and down-regulated respectively, in mice fed with high fat diet compared with the control. The differentially expressed genes were mainly related to free radical oxidative stress, DNA repair, induction of apoptosis, transport, signal transduction and inflammation immune response. Conclusion High fat diet may widely modulate the expression of many genes in the intestine in mice.

Objective To construct a recombinant fusion protein with pneumococcal surface pro-tein A (PspA) of Stretococcus pneumonia (SPN) familyⅠclade 1 and 2, and to analyze the immunogenici-ty of the fusion protein.Methods The gene fragments encoding theα-helix of PspA of the two clades were amplified by PCR and then inserted into the expression vector pET-27b(+) to construct the recombinant ex-pression plasmid.The transformed Escherichia coli BL21 strains carrying expression plasmid were induced by IPTG to express the recombinant protein.The titers and affinity of antibodies against PspA protein were measured by ELISA.An opsonophagocytic assay and an animal experiment were performed to evaluate the immunogenicity of the recombinant protein.Results Double enzyme cutting and gene sequencing confirmed the two purpose gene fragments were correctly expressed in the expression vector pET-27b(+).The titers of anti-PspA antibody in the serum of Kunming ( KM) mice immunized with the fusion protein were 1 ×104 . The affinity of anti-PspA antibody reached to 2×105 .The rates of recombinant PspA6B-PspA05 protein me-diated phagocytosis for SPN6B, SPN05 and SPN01 strains were 20%, 15% and 8.8%, respectively.No SPN23F strain was engulfed by macrophages upon the stimulation with PspA6B-PspA05 protein.The survival rates of mice injected with SPN05, SPN6B, SPN01 and SPN23F strains were respectively 75%, 92%, 75%and 33%upon the immunization of PspA6B-PspA05 protein.Conclusion The recombinant fusion protein PspA6B-PspA05, constructed with the PspA proteins of Stretococcus pneumonia familyⅠclade 1 and 2, was successfully expressed in the E.coli prokaryotic system with the advantage of high immunogenicity.High ti-ters of anti-PspA antibodies with high specificity were induced in KM mice upon the stimulation with Ps-pA6B-PspA05 protein.Moreover, a cross-protective immunity was induced in KM mice upon the immuniza-tion with PspA6B-PspA05 protein.

Objective To explore the disruption of circadian rhythm induced by the alteration of il?lumination leads to cognition impairement and mood dysreguation in mice. Methods 36 male C57BL/6 mice were divided into constant light group( CL) ,normal light group( N) and constant darkness group( CD) . Open field test was applied for the comparison of locomotor activity,tail suspension test and forced swimming test were conducted for assessment of mood state,and elevated plus?maze and Morris water maze were con?ducted for assessment of cognitive function. Results ( 1) Different circadian rhythms did not change the lo?comotor activity among three groups (CL:(200 160.00±955.28)cm,N:(208 148.00±578.11)cm,CD:(179 128.00±1 185.80)cm, P>0.05). (2) Compared with the control group,CL and CD mice showed significantly decreased immobility time in both TST (CD:(40.16±3.82)s,N:(18.83±2.27)s,CL:(46.00±2.80)s, P<0.01) and FST(CD:(181.33±9.03)s,N:(118.83±7.68)s,CL:(151.83±3.06)s, P<0.05). (3) Compared with the control group,CL and CD mice spent less time (CD:(21.76±6.88)s,N:(80.67±11.19)s,CL:(12.50±5.23)s, P<0.05) and made fewer entries (CD:3.33±0.49,N:6.83±0.91,CL:2.00±0.77, P<0.05) into open arms in elevated plus?maze, and exhibited less crossings in target quarter in Morris water maze (CL:2.67±0.76,N:5.00±0.26,CD:2.83±0.40, P<0.05). Conclusion Chronic constant light or darkness leads to negative impacts on mood and cognition in mice.

Objective:To investigate the clinical efficacy of oblique lateral interbody fusion (OLIF) combined with lateral plate (LP) fixation and posterior lumbar interbody fusion (PLIF) combined with pedicle screw fixation in the treatment of adjacent segment disease (ASDis).Methods:Data of 21 ASDis patients treated with OLIF-LP from August 2016 to October 2019 were selected, including 9 males and 12 females; age was 59.3±7.0 years (range, 46-71 years). Target segments: L 2, 3 1 cases (4.8%), L 3, 4 16 cases (76.2%), L 4, 5 4 cases (19.1%). Twenty-one ASDis patients matched with age, sex and surgical segment and treated with PLIF were selected as the control group. The operation time, intraoperative bleeding, postoperative hospital stay, visual analogua scale (VAS), Oswestry disability index (ODI), disc height (DH), intervertebral foramen height (IFH) and lumbar lordosis (LL) were compared between the groups were tested by t-test. VAS score, ODI, DH, IFH and LL were compared within the group by ANOVA, and Bonferroni's test was used for pairwise comparison. Results:All of 42 patients were followed up for 23.7 ±7.4 months (range, 12-36 months). The operation time (97.6 ± 18.0 min) and interpretative bleeding (38.5±62.7 ml) in OLIF-LP group were significantly lower than those in PLIF group (operation time 154.6±42.4 min) and interpretative bleeding (288.6±55.3 ml). There were significant differences between two groups ( t=5.66, P<0.001; t=8.23, P<0.001); the postoperative hospital stay 4.4±1.3 d in OLIF-LP group was longer than that in PLIF group 5.1±1.2 d, but there was no significant difference ( t=1.93, P=0.061); VAS score in OLIF-LP group at 1 month and 3 months after operation (1.6 ± 0.9 points, 1.4 ± 0.8 points), and the ODI index (29.4%±4.7%) after one month operation was improved better than that of PLIF group ( t=2.48, P=0.017; t=2.35, P=0.024; t=2.28, P=0.029), but there was no significant difference between the 12 months after operation of two groups ( t=0.99, P=0.329; t=0.86, P=0.395). The immediately after operation, 3 months after operation and 12 months after operation of DH, IFH and LL in the two groups were significantly improved compared with those before operation ( P<0.05). The immediately after operation, 3 months after operation and 12 months after operation of DH and IFH in the OLIF-LP group were better than those in the PLIF group ( P<0.05), while LL had no significant difference ( P>0.05). There were 2 cases (9.52%) in each group with cage sinking, but no clinical symptoms occurred. In the OLIF-LP group, there was no injury of blood vessels, nerves or abdominal organs during operation, and 2 patients had transient lower limb pain after operation; In the PLIF group, 2 cases (9.52%) of dural rupture were repaired during operation, and no cerebrospinal fluid leakage occurred after operation; Postoperative lower limb pain was aggravated in 3 cases, and improved after dehydration, anti-inflammatory and analgesic treatment; 2 cases of incision exudation healed after symptomatic treatment. Conclusion:OLIF combined with LP fixation has the same clinical effect as PLIF in the treatment of lumbar ASDis, but OLIF combined with LP fixation has more advantages in surgical trauma, postoperative recovery and related complications.

OBJECTIVETo investigate the safety, feasibility and long-term outcomes of laparoscopic complete mesocolic excision for the transverse colon cancer.

METHODSClinical data of 61 patients who underwent laparoscopic complete mesocolic excision for transverse colon cancer (transverse group) in our department from January 2011 to January 2014 were retrospectively analyzed, which were compared with those of 155 patients undergoing laparoscopic complete mesocolic excision for ascending colon cancer (ascending group) and 230 patients undergoing laparoscopic complete mesocolic excision for sigmoid colon cancer (sigmoid group). Differences in operative details, postoperative recovery, postoperative complications and long-term survival among 3 groups were evaluated.

RESULTSNo significant differences in the baseline information were found among 3 groups(all P>0.05). The average operative time was significantly longer in transverse group as compared to ascending group and sigmoid group [(192.1±58.7) min vs. (172.2±54.7) min and (169.1±53.6) min]( P<0.05), while the blood loss [(89.7±63.6) ml, (86.3±66.3) ml, (82.6±61.5) ml], conversion rate [3.3%(2/61), 2.6%(4/155), 2.2%(5/230)], number of harvested lymph node (13.0±4.7, 14.4±6.5, 13.4±5.6), time to flatus [(2.7±1.1) d, (2.6±1.1) d, (2.5±1.0) d], time to liquid diet [(3.0±1.7) d, (2.8±1.5) d, (2.7±1.4) d], incidence of postoperative complication(6.6%, 9.0%, 11.7%), and hospital stay [(11.6±5.8) d, (10.7±5.8) d, (10.6±5.7) d] among 3 groups were not significantly different (all P>0.05). A total of 436 patients received postoperative follow-up of median 36 (5 to 67) months. The overall 5-year survival rate was 73.1%, 73.7% and 74.8%, and the 5-year disease-free survival rate was 71.5%, 71.1% and 72.7% in transverse, ascending and sigmoid colon cancer groups respectively, whose differences were not significant among 3 groups (all P>0.05).

CONCLUSIONLaparoscopic complete mesocolic excision for transverse colon cancer is safe and feasible with slightly longer operation time, and has quite good long-term oncologic efficacy.

Colectomy ; methods ; Colon, Ascending ; surgery ; Colon, Sigmoid ; surgery ; Colon, Transverse ; surgery ; Colonic Neoplasms ; surgery ; Comparative Effectiveness Research ; Disease-Free Survival ; Humans ; Laparoscopy ; methods ; Length of Stay ; Lymph Node Excision ; Lymph Nodes ; Mesocolon ; surgery ; Operative Time ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Survival Rate ; Treatment Outcome

A reverse-phase high pressure liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) has been developed for the quantitative analysis of hupehenine in the total alkaloids from Fritillaria hupehensis. Samples were analyzed on a reverse-phase column (Hypersil C-18) with a mobile phase of methanol:water:chloroform: triethylamine (85:15:1:0.6). The ELSD was set at the drift tube temperature of 68.3℃ and gas flow rate of 1.8 L/min. Hupehenine's retention time was 13.7 min with an asymmetry factor of 1.2. The validity of the method has been verified with linearity, limit of detection, accuracy and precision. The logarithmic linear curve was obtained from 8.936 to 134.04 μg/mL (r=0.9993). The detection limit (S/N＞3) of hupehenine was 1.79 μg/mL on the column. Intra-day RSD was 1.42% and inter-day RSD was 2.26% (3 days within a week). The average recovery of hupehenine was 101.50%, and RSD was 1.62%.