BACKGROUND:DCLK1 is a transmembrane microtubule-associated kinase in neurons after mitotic division, which may be the intestinal cancer stem cel marker. OBJECTIVE:To observe the expression and pathological significance of DCLK1 and Ki67 in colorectal cancer. METHODS: Expression of Ki67 and DCLK1 in 150 cases of colorectal cancer tissues was detected by immunohistochemical method in contrast to normal colorectal mucosa, para-carcinoma tissue, and adenoma tissue. RESULTS AND CONCLUSION:The expression rates of DCLK1 and Ki67 were 36.7% and 34.7% in cancer tissues, respectively, both of which were significantly higher than those in normal colorectal mucosa and adenoma. The expression of DCLK1 was associated with the location, depth of invasion, lymph node metastasis (P < 0.05), while the expression of Ki67 was just associated with the depth of invasion (P < 0.05). There was a negative correlation between the expression of DCLK1 and Ki67 (r=-0.460,P=0.000). The count of DCLK1+/Ki67-cels was about 2.01% in colorectal cancer tissues, and these cels mainly distributed at the bottom of intestinal mucosa base and common duct wal. DCLK1+/Ki67- cels were oval, the nuclei were large and deep-stained with prominent nucleolus, and there was rare nuclear fission and less cytoplasm. From the aspects of cel number, location, and cel morphology, DCLK1+/Ki67- cels are in line with the characteristics of cancer stem cels; therefore, DCLK1+/Ki67-can be used as a cancer stem cel marker of colorectal cancer.

Objective To investigate clinical characteristics of tuberous sclerosis(TSC)without nervous system abnormalities in infants. Methods Clinical manifestations of 12 infants with TSC were analyzed. Reflectance confocal microscopy (RCM)was performed in 5 of the 12 infants. Two patients underwent histopathological examination and electron microscopic examination. Results The age at first clinic visit for TSC ranged from 2 to 18 months (median, 8.6 months)among these patients. Before occurrence of nervous symptom, TSC mainly manifested as hypomelanotic macules, nodules in the subependymal layer or walls of lateral ventricles of the brain, cardiac rhabdomyoma and multiple renal cysts. RCM revealed intact dermal papillary rings with low reflectivity and obscure boundaries in 5 infants. Histopathological examination showed a local decrease in melanocytes and pigment granules in basal cells. Electron microscopic examination showed a normal number of melanin granules in the cytoplasm of melanocytes in the basal layer. Conclusions Before appearance of nervous system abnormalities, TSC mainly manifests as hypomelanotic macules in infants, and is easily confused with vitiligo in clinic. RCM examination may serve as a useful method to distinguish infant TSC from hypopigmented diseases.

Objective To build a set of inexpensive wireless wide area network PACS based on open source code.Methods To improve the open source code MyFreePACS on Internet.Results The basic function of PACS can be achieved in an economical way,using internet explorer to browse and deal with the DICOM Images,with the capability of such functions as displaying and managing of the pictures.It is possible to make rounds at patient bedside with wireless device and remote medical consultation with specialists through Internet.Conclusion It is possible to utilize the free open source code MyFreePACS on the internet to construct an economical PACS on Web in the medium and small-scale hospital.

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

Professor LI Zhi-dao's acupuncture technique at muscle belly and its clinical experiences for the treatment of arthralgia are introduced. From aspects of TCM, anatomical physiology and sports biomechanics, the theory basis of this technique is explained, while its method is elaborated in details from acupoint selection and manipulation combined with classic medical cases. The acupuncture technique at muscle belly is applied at the muscles which already have pathological changes, avoiding re-stimulation on the pain area. It provides new thinking methods and references for acupuncture to treat arthralgia.
Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease ( TALEN) mRNAs.Methods TALEN knockout site recognizing Txnip was designed by tools on line, then constructed the vectors and assayed its cleavage activity at cellular level.TALEN mRNA was transcribed in vitro and microinjected into C57BL/6J mouse zygotes.F0 mice were verified at DNA level with BamHI and TXNIP-knockout mice were obtained.Results We designed and constructed TALENs which recognized and cut the first exon of Txnip, and got four TXNIP knockout mice, among which two were frameshift mutation, demonstrating that the TXNIP-knockout mice were generated by TALEN technique.Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

OBJECTIVETo research serum level of interleukin-17 (IL-17) associated with the progression of hepatic injury in the chronic patients with hepatitis B.

METHODSThe serum level of IL-17 was measured by ELISA and the serum levels of IL-6, IL-8 were measured by RIA in patient groups and healthy group, the patient groups including 42 mild patients, 37 moderate patients and 38 severe patients.

RESULTSIL-17,IL-6 and IL-8 levels in patient patients were significantly higher than those in healthy people (P < 0.01). There is no significant difference among mild patients and moderate patients. Compared with mild patients and moderate patients,the cytokines lever were significantly higher in severe patients (P < 0.01).

CONCLUSIONIL-17 as a new cytokine probably play a multiple role as immune factor and inflammation element in the progression of the chronic hepatitis B disease. Maybe, it can provide a new approach to the therapy of the chronic hepatitis B.

Adult ; Aged ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; blood ; pathology ; Humans ; Interleukin-17 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged

Abstract: Objective　To understand the general situation of death causes of permanent residents in Hainan Province from 2014 to 2020, analyze the causes of death, and provide scientific basis for the formulation of policies related to healthy Hainan. Methods The Hainan Provincial Population Death Information Registration and Management System was used to report data, and the death information of permanent residents in Hainan Province from 2014 to 2020 was obtained. Excel 2010 and SPSS 26.0 software were applied to perform statistical analysis related to the indicators including crude mortality rate, standardized mortality rate, ranking of causes of death, and composition ratio; the standardized mortality rate was calculated based on the data of the 2010 National Census Data. Results　From 2014 to 2020, the average annual resident population of Hainan Province was 9 175 300, and average annual resident population at each of the eight monitoringp oints is 297 100， anaverage of 83 878 cumulative deaths were reported, the annual total mortality rate was 504.09/100 000, the standard mortality rate was 618.69/100 000. The standardized mortality rates of male and female were 806.18/100 000 and 444.36/100 000 respectively (P<0.01). The mortality rate of all age groups showed that the crude mortality rate of 0~<1 years old group showed a decreasing trend with the passage of time, and the crude mortality rate was significantly higher than that of other age groups. From the age of 20, with the increase of age, the overall crude mortality rate of residents keeps increasing. The crude death rate increases rapidly after the age of 65, and peaks especially after the age of 85. The leading causes of death were circulatory diseases, tumors and respiratory diseases. From 2014 to 2020, the mortality rate of circulatory diseases and tumors, the two main causes of death, was more than 100/100 000. Conclusion　Chronic non-communicable diseases are still the first cause of death among permanent residents in Hainan Province. Targeted intervention and treatment of chronic non-communicable diseases is beneficial to reduce their mortality.

A two-dimensional liquid chromatography method was developed for the analysis of rice leaves proteomics based on the coupling of hydrophilic interaction liquid chromatography-reversed-phase liquid chromatography with online tandem mass spectrometry. The influence of pH value of chromatographic mobile phase on the orthogonality of the hydrophilic interaction-reversed-phase two-dimensional liquid chromatography was evaluated by the changes of standard peptide retention. The results indicated that the better orthogonality (R2=0.34113) was achieved from the system with hydrophilic interaction columns(pH 9.3) in the first and C18columns(pH 3.3) in the second LC dimension. Coupled with multiple fraction concatenation strategy,the orthogonality of two-dimensional liquid chromatography was further evaluated in the analysis of complex rice leaf proteins. The results showed that more than 50% of the total peptides were identified less than two times, and the peptides obtained from first-dimension were well distributed across the elution window,indicating that the method showed significant orthogonality in the identification of complex rice leaf proteins. Based on the proteome discoverer software,207345 peptides belonged to 2930 protein clusters were identified.

Objective: To explore the mechanism of herbal cake-partitioned moxibustion in Crohn disease (CD) treatment by observing the effect of herbal cake-partitioned moxibustion on protein expressions of colonic M2 macrophage marker CD206, AMP-activated protein kinase (AMPK) and tuberous sclerosis complex (TSC) 2. Methods: Twenty-six specific pathogen free male rats were randomly divided into a normal group, a model group and a herbal cake-partitioned moxibustion group. The CD model was prepared by enema with the mixture of 5％ (W/V) 2,4,6- trinitrobenzene sulfonic acid (TNBS) and 50％ ethanol at 2:1 (volume ratio). After the model was successfully prepared, rats in the herbal cake-partitioned moxibustion group received herbal cake-partitioned moxibustion at Qihai (CV 6) and bilateral Tianshu (ST 25). Hematoxylin-eosin (HE) staining was used to observe the histopathological changes of rat colon; immunohistochemical technique was used to detect the expression of colonic CD206 protein; Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) technologies were used to detect the protein and mRNA expressions of colonic AMPK and TSC2. Results: Compared with the normal group, rats in the model group showed damaged colonic mucosa, missing of the epithelial layer, thickened submucosa, vascular proliferation, massive infiltration of monocytes and lymphocytes, and cracked ulcers that reached the muscle layer. Rats in the herbal cake-partitioned moxibustion group showed reduced intestinal inflammation and healing intestinal epithelium ulcers. Compared with the normal group, rat colonic CD206 protein expression, and the protein and mRNA expressions of colonic AMPK and TSC2 were decreased in the model group (all P<0.01); compared with the model group, rat colonic CD206 protein expression was increased (P<0.01), as well as the protein and mRNA expressions of AMPK and TSC2 in the herbal cake-partitioned moxibustion (all P<0.05). Conclusion: Herbal cake-partitioned moxibustion can reduce intestinal inflammation in CD rats, increase colonic CD206 protein expression, and up-regulate the protein and mRNA expressions of colonic AMPK and TSC2.