Vitiligo is a kind of difficult to treat skin disease. Its pathogenesis is not very clear and the treatment is also difficult. In this paper, according to basic theories of traditional Chinese medicine (TCM) from basic drug selec-tion, visceral syndrome differentiation, harmonizing qi and blood, expelling wind and eliminating dampness, treatment according to four seasons, medication according to meridian pathways, psychotherapy and nursing. Experiences of professor W ang Jusheng in vitiligo treatment were introduced in order to provide a beneficial reference.

Amounts of Certain elements in Anopheles sinensis, both normal ones (emerged mosquitoes, d0, d5, d8, d12 and d18 after taking blood meal) and those infected with Brugla malayi microfilaria (d5, d 8, d12 and d18 after infection), were determined using IL-551 and PE-603 atomic absorption spectrometer(AAS). The results showed that 9 kinds of trace elements including Fe, Zn, Cu, Cd, Al, Pb, Mn. Ni and Cr and 4 kinds of macro elements as K, Na, Ca, Mg were present in both the noninfected and infected mosquitoes. Comparing the contents of the elements between the noninfected and infected mosquitoes, the amounts of many kinds of elements reduced obviously in mosquitoes infected with microfilaria. The quantity and kinds of elements reduced in mosquitoes with the days of infection, for instance, 10 kinds of elements on the 5th day, 11 on the 8th day, 9 on the 12th day and 7 on the 18th day after infection(Tables 1, 2).

Objective:To study the correlation between the thickness of psoriasis skin and TCM syndromes,and to find an uninjurious and effective way to determine TCM syndromes by high-frequency ultrasound.Methods:Eighty-seven patients were divided into three groups:blood-heat syndrome group,blood-dryness syndrome group and blood-stasis syndrome group.The patients' epidermal and dermal thickness of targeted psoriasis skin and surrounding normal area was measured by high-frequency ultrasonic apparatus.Results:The thickness of epidermal and dermal of psoriasis skin was thicker than that of surrounding skin,and it was more obviouse of derma.The dermal thickness indexes of blood-stasis patients was the biggest in the three groups(P

A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patients with hepatitis B-related hepatocellular carcinoma (HCC) and 3 cases of normal liver tissues. The integration sites and flanking sequences in human genome were sequenced and blasted, and the expression of integrated HBV genes was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The influence of the up-regulated expression of integrated genes on hepatocarcinogenesis was analyzed. Nineteen integration sites of HBV DNA into HCC tissues were obtained by RT-PCR and sequencing. These genes encoding proteins were: LOC51030, LOC157777, minichromosome maintenance complex component 3 associated protein (MCM3AP), MCTP1, SH3 and multiple ankyrin repeat domains 2 isoform 2, CCDC40, similar to HCG2033532, mitochondrial ribosomal S5 pseudogene 4. One of them was integrated into the intron of MCM3AP. RT-PCR demonstrated that the expression levels of MCM3AP mRNA in HCC tissues, matched surrounding liver tissues and normal liver tissues were in a descendent order. The ratio of MCM3AP mRNA to the GAPDH mRNA in these three tissues was 1.07375, 0.21573, 0.06747 respectively, with the difference being statistically significant among them (P<0.05). Meanwhile, the expression levels of MCM3AP mRNA from HCC tissues in which HBV DNA integrated into MCM3AP were still significantly higher than those without HBV DNA integrated into MCM3AP. It was concluded that the HBV DNA integration sites into human genome were random, and MCM3AP was a new site. The up-regulated MCM3AP mRNA may affect flanking sequences which promote the hepatocarcinogenesis.

Objective To find a novel operative modality with sphincter preservation in the treatment of middle and low rectal carcinoma. Methods Pull through lower resection was performed on 28 rectal cancer patients. The distance between the anal verge and the lower margin of the tumor was 6～8cm(20 patients) or 8～10cm(8 patients), including 8 patients in Dukes A stage, 16 Dukes B and 4 Dukes C. The resected line from tumor distal margin was 2cm, 3cm, and 4cm, respectively. Results There was no operative death, anastomotic fistula or anastomotic stenosis in these cases. Mean follow up period was 30 months. Local recurrence was found in two cases (7.1%) 18 months after the operation, and 26 cases were cancer free till the end of the follow up. Defecation was satisfactorily controlled 8～12 weeks after the operation. Conclusions Pull through Welch procedure could meet the criterion of the radical resection of rectal carcinoma,and keep the internal and external sphincter muscles intact in the superior lower anterior resection. The normal defecationcan can maintain after the operation due to the preservation of internal and external sphincter muscles.

Objective : To examine the association between dietary patterns during pregnancy and gestational diabetes mellitus (GDM), so as to provide the evidence for guiding the establishment of healthy and balanced dietary patterns and reducing the prevalence of GDM. Methods: Pregnant women who underwent oral glucose tolerance tests in Hangzhou Obstetrics and Gynecological Hospital from 2020 to 2021 were enrolled, and their demographic information were collected using questionnaires. Pregnant women's diets during the past three months were collected using Food Frequency Questionnaires (FFQs), and dietary patterns were extracted using principal component analysis. In addition, the association between dietary patterns and risk of GDM was examined using a multivariable logistic regression model. Results: A total of 1 689 pregnant women were included, with a median age of 28.53 (interquartile range, 2.47) years and a median gestational age of 26.00 (interquartile range, 2.00) weeks. Five dietary patterns were identified according to pregnant women's types of diets, including meat-based diets, dessert-fruit-refined grain diets, plant-based diets, eggs-milk-nut diets and whole-grain diets, with a cumulative contribution rate of 58.76%. The prevalence of GDM was 24.57% (415 cases) among the study subjects. Multivariable logistic regression analysis showed that pregnant women with scores in the highest quartile (Q4) of the meat-based diets had an increased risk of GDM (OR=1.372, 95%CI: 1.043-2.055) relative to those with scores in the lowest quartile (Q1), and pregnant women with Q4 scores of the dessert-fruit-refined grain diets had an increased risk of GDM (OR=1.743, 95%CI: 1.397-2.432) relative to those with Q1 scores, while pregnant women with Q4 scores of the plant-based diets had a reduced risk of GDM (OR=0.382, 95%CI: 0.346-0.613) relative to those with Q1 scores. Conclusion A plant-based dietary pattern may reduce the risk of GDM, while meat-based and dessert-fruit-refined grain dietary patterns may increase the risk of GDM.

Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.

HBV polymerase has intrinsic RNA-dependent reverse transcriptase, DNA-dependent DNA polymerase as well as RNaseH activity. Analysis of HBV polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. To obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. The gene of HBV polymerase was amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-pol. The recombinant donor plasmid, pFastbac Dual-pol, was constructed by inserting HBV polymerase gene into EcoRI and PstI sites controlled by polyhedrin promoter. The recombinant donor plasmid was transformed into DH10Bac competent cells for transposition. Recombinant bacmid was constructed by inserting of the mini-Tn7 element from the donor plasmid into the mini-attTn7 attachment site on the bacmid. The recombinant bacmid DNA was isolated and transfected into the Sf9 cells to produce the recombinant virus, and healthy insect Sf9 cells were infected with the recombinant virus containing HBV polymerase gene to express the target protein. HBV polymerase expressed in insect cells was analyzed by SDS-PAGE. PCR results showed recombinant donor plasmid, pFastbac Dual-pol, was constructed successfully. The recombinant hepatitis B virus polymerase was expressed in insect cells at high level. The recombinant hepatitis B virus polymerase should facilitate the analysis of HBV polymerase biological characteristics, allow the investigation for new anti-HBV drugs specifically blocking HBV polymerase.

Objective To investigate the relationship between single nucleotide polymorphism (SNP) of Fas ligand (FasL) and fulminant hepatitis B in Han Chinese. Methods HBV infected subjects were enrolled in this case-control study, including 233 cases of inactive HBsAg carrier, 68 patients with fulminant hepatitis B,100 cases of spontaneous hepatitis B clearance, 102 patients with hepatitis B virus (HBV) related cirrhosis and 112 patients with HBV related primary hepatocellular carcinoma. The blood samples and clinical data were collected. FasL-844T/C polymorphisms of enrolled subjects were examined by TaqMan real time fluorescent genotyping polymerase chain reaction (RT-PCR). A adjusted odds ratios (OR)and 95% confidence intervals (CI)were calculated using the Logistic regression model. Results After adjusting the factors of gender and age, binary Logistic regression analyses indicated that the genotype frequencies of FasL-844 CC,CT,TT in inactive HBsAg carriers were 50. 64% ,39. 91% and 9. 44% respectively, and those in cases of fulminant hepatitis B were 79. 41%, 17. 65% and 2. 94%, respectively. The analysis also revealed that FasL-844CC genotype in inactive HBsAg carriers was high risk factor of developing fulminant hepatitis B (OR =4. 729,95%CI:0. 510 - 21. 282,P = 0. 043), while there were no statistic significances in other cases (P>0. 05). Conclusion The inactive HBsAg carriers harboring FasL-844CC may have greater susceptibility to fulminant hepatitis B, which need arouse high attention.

BACKGROUND: Human keratinocyte growth factor-2 (hKGF-2) has extensive physiological functions, which plays an important role in embryonic development, tissue-repairing, nervous regeneration, vascularization and development of tumor.OBJECTIVE: To clone hKGF-2 gene, obtain the expression of hKGF-2 in Escherichia coli(E.coli) and determine its bioactivity, so as to provide experimental basis for further investigation.DESIGN: Open experiment.SETTING: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention.MATERIALS: The experiment was conducted in State Key Laboratory of Viral Genetic Engineering, Institute for Viral Disease Control and Prevention of Chinese Center for Disease Control and Prevention. The temperature control expression vector pBV220 was constructed by State Key Laboratory of Viral Genetic Engineering; EcoR Ⅰ , BamH Ⅰ , T4 DNA ligase (Promega Co., Ltd.); The specific polymerase chain reaction (PCR) of hKGF-2 (Manufactured by Shanghai Boya Biotechnology Co., Ltd.); Heparin-Sepharose CL-6B (Pharmacia Company); PCR rapid purification kit,Trizol kits for total RNA extract, Kits for RT-PCR (GIBCO Co., Ltd.); Kits for rapid extraction of plasmid DNA (Boda Company); BL-21-codon plus compent cells (Stratagene Co., Ltd.).METHODS: High-expression strain BL-21 codon plus competent cells was used to express and purify initially recombinant hKGF-2 protein, and its activity was detected. RT-PCR was adopted to obtain hKGF-2 cDNA from lung tissues of naturally aborted fetus and clone it into pBV220 carri er plasmid. The hKGF-2 protein expressed in BL-21 codon plus competent cells of E.coli. Affinity chromatography and ion exchange chromatography were applied in isolation and purification, and the bioactivity of expression protein was determined in cell proliferation test.MAIN OUTCOME MEASURES: The length and sequence of cDNA segment in hKGF-2, the expression of hKGF-2 gene inE.coli and the purification of hKGF-2 activity.RESULTS: The segment of hKGF-2 cNDA was about 500 bp, and hKGF-2 protein highly expressed in BL-21, which had soluble expression in the supernatant. SDS-PAGE showed that the relative molecular mass was about 20000, and hKGF-2 protein could significantly promote the mitotic activity of NIH3T3 cells. The A value (490 nm) of hKGF-2 in the 1 μg/L, 5 μg/L and 10 μg/L groupswere higher than that in the blank control group, and the differences were significant (which were 0.174±0.022,0.220±0.029,0.306±0.050,0.066±0.004 respectively,P ＜ 0.001).CONCLUSION: hKGF-2 gene is successfully cloned, which highly expresses in BL-21 of the E.coli. Purified hKGF-2 protein can stimulate the proliferation of NIH3T3 cells and significantly promote its mitotic activity.