Zinc supplementation can reduce the mean duration,the recurrence rate and mortality of diarrhea.Nonetheless,it has no effect on stool frequency or stool output.Zinc supplementation results in different effort for the difference of the duration of diarrhea,pathogens,zinc type and doses of zinc,and shows no effect in infants younger than 6 months.Using iron as a co-intervention can reduce the effect of zinc,but vitamin A or ORS have a synergistic effect with the meeting.Average baseline zinc levels did not contribute to variations in the effect size.Zinc supplements are unlikely to improve compliance with the treatment for diarrheal disease.Longer course,zinc alone,malnutrition,diarrhea,breast-feeding children are prone to vomiting after oral administration of zinc.

Objective To explore the distribution pattern of G protein-coupled receptor family C, group 6, subtype A (GPRC6A) mRNA in adult mice. Methods The distribution of GPRC6A mRNA in paraffin embedded adult mouse tissues was determined by highly sensitive nonradioactive cRNA probe in situ hybridization (ISH). We compared ISH with and without addition of tyramide signal amplification (TSA). GPRC6A wild-type and littermate GPRC6A null mice tissue sections were investigated by ISH. Results TSA greatly increased the sensitivity of ISH to detect GPRC6A mRNA in wild type mouse tissues. There was no detection of GPRC6A mRNA in GPRC6A gene specific knockout tissue in paraffin embedded tissue section. The mRNA of GPRC6A was detectable in the digestive gland or accessory digestive gland including salivary gland and pancreas, as well as in the tissues including kidney, testis, brain, muscle, and fat. Conclusion The mRNA distribution pattern of GPRC6A gene is compatible with the phenotype of GPRC6A knockout mice.

BACKGROUND: Stromal cell-derived factor 1 (SDF1), a potential inhibitor of infection by lymphophilic HIV-1 strains, can help to block the pathway of HIV-1 invasion into the human body.OBJECTIVE: Genotype and polymorphism of SDF1-3 'A allele associated with HIV-1 infection were investigated in She Ethnic Group in the south of China so as explore the possible causes of uninfection by HIV-1 strains among this population.DESIGN: Single sample study.SETTING: Department of Biochemistry and Molecular Biology, Gannan Medical College.PARTICIPANTS: Totally 186 She Ethnic subjects without HIV-1 infection collected randomly from those whose three generations belonged to She Ethnic Group, and inhabited in Qianshan County of Jiangxi Province,Ningde area of Fujian Province and Jingning She County of Zhejiang Province, from January to December 1995.METHODS: The whole blood samples from 186 She Ethnic subjects were collected randomly, and then their genomic DNA samples were extracted respectively. Allelic polymorphism was examined by the polymerase chain reaction and restriction-fragment-length polymorphisms (PCR-RFLP).MAIN OUTCOME MEASURES: The distribution of SDF1-3'A allele in She Ethnic Group in the south of China.RESULTS: The data of 186 She Ethnic subjects entered the result analysis without any loss in the midway. The frequency of SDF1-3 'A allele in She Ethnic Group samples was 19.6%, and the allelic distribution of the gene was in accordance with Hardy-Weinberg equilibrium. No difference was found between male and female individuals.CONCLUSION: The frequency of SDF1-3 'A allele of She Ethnic Group in the south of China was similar to that of Dai Nationality in Yunnan.Based on its slow-down effect on clinical course of AIDS, the mutation of SDF1-3'A is significant in the prevention and treatment of AIDS in She Ethnic Group in the south of China.

Objective To investigate the effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages by analyzing the expression of lincRNA-cox2 in RAW264. 7 macrophages of M1 and M2 phenotypes. Methods Murine RAW264. 7 cells were induced by IFN-γand LPS to polarize to M1 phenotype, and were induced by IL-4 to polarize to M2 phenotype. The expression of lincRNA-cox2 in M1 and M2 macrophages were analyzed by real-time quantitative PCR ( RT-PCR) . We designed and synthesized siRNA oligo for lincRNA-cox2 and unrelated sequences. Then the siRNA oligo and NC oligo were transfected into RAW264. 7 cells by LipofectmineTM 2000. The transfected RAW264. 7 cells were induced by IFN-γand LPS or by IL-4 to polarize to M1 or M2 macrophages. Enzyme linked immunosorbent assay ( ELISA) was performed to measure the secretion of IL-10 and IL-12 induced in different conditions. The expression of in-ducible nitric oxide synthase ( iNOS ) , TNF-α, arginase 1 ( Arg-1 ) and found in inflammatory zone 1 (Fizz1) at mRNA level were detected by RT-PCR. The M1 macrophages were transfected with siRNAs to knock down the expression of lincRNA-cox2 for analyzing the biological effects of lincRNA-cox2 on the polar-ization of macrophages. Results The relative expression of lincRNA-cox2 in M1 macrophages was signifi-cantly higher than that in RAW264. 7 cells and M2 macrophages. Compared with the control group, the RAW264. 7 cells transfected with lincRNA-cox2-siRNA showed decreased secretion of IL-12 and inhibited expression of iNOS and TNF-αat mRNA level after IFN-γand LPS induction, but increased secretion of IL-10 and enhanced expression of Arg1 and Fizz1 at mRNA level after IL-4 induction. Transfecting the M1 mac-rophages with lincRNA-cox2-siRNA inhibited the secretion of IL-12, but promoted the secretion of IL-10. Conclusion This study indicated that lincRNA-cox2 was involved in the regulation of macrophage pheno-types by promoting the polarization to M1 macrophages and inhibiting the polarization to M2 macrophages.

Objective To construct a recombinant bacterial vaccine which can express specific 10-23 deoxyribozyme(DZ) in macrophage, identify the intracellular production of specific 10-23DZ and detect the activity of this recombinant bacterial vaccine on inhibiting the expression of TACO gene in macrophage.Methods The pSDE02 was obtained by inserting the replicon of Mycobacterium into pSDE01, a plasmid which can express 10-23DZ in eukaryotic cells. The expression sequence of DZ1, a 10-23DZ targeting the TACO mRNA of macrophage designed in our previous study was synthesized and inserted into pSDE02. The resulted plasmid was named pDZM01. pDZM01 was then transferred into Mycobacterium smegmatis by electroperation. The recombinant M. smegmatis, named rMs-DZ1 was screened on low-salt LB medium containing Zeocin and identified by Colony PCR. The targeted delivery property of recombinant M. smegmatis was observed by Ziehl-Heelson stain and GFP expression observation via fluorescence microscope. rMs-DZ1 was used to infect RAW264.7 cells and the expression of DZ1 in macrophage was identified by dot-blot assay. At 24 h and 48 h after infection, total RNA and proteins were extracted and the TACO mRNA and protein expression level was assayed by RT-PCR and western-blot respectively. Results Restrictive analysis and sequencing data showed that the Mycobacterium-eukaryotic cell shuttle plasmid pSDE02 and pDZM01 was successfully constructed. rMs-DZ1 was confirmed by colony PCR. When engulfed by macrophage, rMs-DZ1 would express DZ1 in RAW264.7 cells and inhibit the expression of taco gene. When compared to uninfected macrophage, rMs-DZ1 significantly reduced the taco mRNA by 67.90% and 57.14% and down-regulated the expression of TACO protein by 53.85% and 68.92% at 24 h and 48 h respectively. Conclusion A recombinant M. smegmatis vaccine was successfully constructed which could generate specific 10-23DZ in macrophage and inhibit the expression of target gene of interest. To our knowledge, this is the first bacterial vector which can express intracellularly 10-23DRz in targeted manner. This study may further prompt the feasibility of using 10-23 DNAzyme to achieve effective and targeted gene silence.

Objective To evaluate the diagnostic value of dual-source CT angiography (DSCTA) for intracranial aneurysms.Methods The data of DSCTA and digital subtraction angiography (DSA) were collected from 95 patients with subarachnoid hemorrhage (SAH).The efficacies of detection and description of morphologic features of intracranial aneurysms were analyzed retrospectively.Results A total of 117 aneurysms in 88 patients were detected with DSCTA.Two patients were suspected of having aneurysms,and no aneurysrms were detected in 5 patients.These patients were reexamined with DSA,4 were diagnosed as having aneurysm,and the aneurysms were not detected in 3 patients.DSA results were considered as gold standard,the specificity,sensitivity and accuracy of DSCTA for the detection of intracranial aneurysms were 100％,96.7％and 96.8％,respectively.The larger volume of intracranial aneurysm was,the higher the sensitivity of DSCTA diagnosis would be.Even for small aneurysms,the sensitivity of DSCTA diagnose was more than 90％.In addition,tmeasurement results of the maximum diameter and neck width of aneurysms measured by DSCTA were almost consistent with DSA.Condclusions SCTA is a non-invasive,quick,reliable,and effective method,and can provide accurate imaging information for surgery.The specificity and sensitivity of the diagnosis of aneurysms with DSCTA are almost the same with DSA.It has more advantages than DSA in the emergency operation of intracranial aneurysms.

Objective To construct a lentiviral vector-based short hairpin RNA (shRNA) targeting the gene encoding tryptophan-aspartate containing coat protein ( TACO) and to evaluate its inhibitory effect on the expression of TACO , and to further elucidate its effects on the phagocytosing and intracellular killing of My-cobacterium tuberculosis (M.tb) by macrophages and the possible mechanisms.Methods Three shRNA frag-ments targeting TACO gene and a scrambling control shRNA fragments were designed and cloned into the lentivi -ral vector pSicoR .The recombinant lentiviral vectors were identified by sequencing analysis and then packed in 293T cells.Real-time RT-PCR and Western blot assay were performed to evaluate the gene-silencing efficiency of the recombinant lentiviral vectors among RAW 264.7 cells transfected with the concentrated lentivirus .The most effective lentivirus was screened out to transfect the RAW 264.7 cells for 48 hours, followed by infection those cells with M.tb strains.The entry and intracellular survival of M .tb strains in RAW264.7 cells were de-termined by bacterial culture at indicated time points .The colocalization of M .tb and lysosomes was detected by immunofluorescence staining .The cyto-ID autophagy kit was used to detect the cellular autophagy and the auto-phagy-associated protein LC 3 was determined by Western blot assay .Results The recombinant lentiviral vec-tors were successfully constructed and confirmed by sequencing analysis .Decreased expression of TACO in RAW264 .7 cells was detected after transfecting the cells with the lentiviral vector-based shRNA vectors targeting TACO gene for 48 hours.The most effective lentivirus , LV-pSRT1, decreased the expression of TACO by 85.24%and 69.00%at the mRNA and protein levels, respectively.The bacterial loads in LV-pSRT1 trans-fected RAW264.7 cells were significantly decreased at the time point of 0 h after M.tb infection as compared with those in the control lentivirus treated cells (5.50×104 vs 8.1 ×104, P<0.05).Compared with the RAW264 .7 cells transfected with control lentivirus , the survival rate of intracellular M .tb strains in LV-pSRT1 transfected cells was significantly decreased at the time point of 48 h (134.54% vs 213.58%, P<0.05) and 72 h (148.18%vs 262.96%, P<0.05) considering the bacterial load at the time point of 0 h as the standard. The immunofluorescence staining demonstrated that the colocalization of M .tb strains with lysosomes was signifi-cantly enhanced in LV-pSRT1 transfected cells as compared with that in control lentivirus treated cells (75.67%vs 10.66%, P<0.05).Moreover, significantly enhanced autophagy and relative expression of LC 3Ⅱ protein were observed in RAW264.7 cells with TACO gene knockdown (16.20%vs 8.50%, P<0.05;0.51 vs 0.34, P<0.05).Conclusion The lentiviral vector-based shRNA targeting TACO gene could effectively knockdown the expression of TACO protein , decrease the entry and increase the intracellular killing of M .tb strains in mac-rophages.The enhanced intracellular killing of M .tb strains by macrophages was associated with the increased fusion of M.tb-containing phagosome and lysosome .

Objective To evaluate the short-term efficacy of recombinant human endostatin (rhES) combined with transcatheter arterial chemoembolization (TACE) versus TACE alone for intermediate and advanced primary hepatic carcinoma.Methods The relevant controlled trials about rhES plus TACE versus TACE alone in the treatment of intermediate and advanced primary hepatic carcinoma were retrieved from the databases of PubMed,Elsevier,Cochrane Library,China National Knowledge Infrastructure (CNKI),Chinese Biomedical Literature Database (CBM),Wan Fang Database.The retrieval time limited was from the database construction to January 2018,and the Meta-analysis was performed by using RevMan5.3 software.Results 18 controlled trials were included in this Meta-analysis.There were 948 cases,of which 522 cases in rhES plus TACE group and 426 cases in TACE alone group.According to the usage of rhES,the trials were further divided into intrahepatic arterial embolization group,intrahepatic arterial pump group,and intravenous infusion group.rhES plus TACE had an overall advantage over TACE alone in terms of objective response rate (ORR),and the difference was statistically significant (RR =1.59,95％CI:1.41～1.79,P＜0.05).And the ORR of rhES plus TACE in intrahepatic arterial embolization group,intrahepatic arterial pump group,intravenous infusion group was better than that of TACE alone (Intrahepatic arterial embolization group RR=1.63,95％CI:1.36～ 1.95;Intrahepatic arterial pump group RR=1.49,95％CI:1.24～1.79;Intravenous infusion group RR=1.69,95％CI:1.22～2.34),and the difference was statistically significant (P＜0.05).The subgroups analysis of anthracycline and platinum also showed that ORR in rhES plus TACE patients was better than that in TACE patients alone.Conclusion The short-term efficacy of rhES plus TACE in the treatment of intermediate and advanced primary hepatic carcinoma was better than that of TACE alone,and the same results were obtained by subgroup analysis.

Objective To evaluate the efficacy of Disposcope endoscope(DE)-guided nasotrache-al intubation in patients with difficult airway by comparing with fiberoptic bronchoscope(FOB). Methods One hundred and twenty American Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both se-xes, with body mass index<25 kg∕m2, aged 18-64 yr, with mouth opening<3 cm, of Mallampati classifi-cation Ⅲ or Ⅳ, undergoing maxillofacial surgery requiring nasotracheal intubation were divided into DE group(n=60)and FOB group(n=60)using a random number table.Nasotracheal intubation was per-formed under the guidance of DE or FOB after induction of anesthesia.Glottis exposure was evaluated using Cormack-Lehane grade.Epistaxis during intubation, successful intubation, time and degree of glottis expo-sure, intubation time and development of tachycardia and hypertension and requirement for assisted ventila-tion with face mask during intubation were recorded.The patients were followed up postoperatively, and the development of intubation-related complications was also recorded.Results Compared with group FOB, glottis exposure time and incubation time were significantly shortened(P<0.05), and no significant change was found in Cormack-Lehane grade, rate of successful incubation, rate of successful intubation at first attempt or intubation-related complications in group DE(P>0.05). Hypertension and tachycardia were not found and no patients required assisted ventilation with face mask during intubation in the two groups.Conclusion DE-guided nasotracheal intubation provides similar efficacy to that of FOB with shorter time and is an optimal selection for the patients with difficult airway.