Objective:To construct plasmid coexpressing GM-CSF and HBsAg,enhance hepatitis B DNA vaccines antivirus effect.Methods:(1)The HBsAg-encoding fragment and GM-CSF-encoding fragment were created by PCR amplification from pEcob6 and pCD-hGM-CSF respectively,then were cloned into plasmid pcDNA3.1(+),pcDNA3.1-S and pcDNA3.1-GM-CSF(without stop codon)were constructed.(2)HBsAg DNA fragment (with stop codon) was amplified by PCR,then was subcloned into plasmid pcDNA3.1-GM-CSF.Results:Recombinant pcDNA3.1-S and pcDNA3.1-GM-CSF-S were confirmed by using restriction enzymes and DNA sequencing.HBsAg was detected in the lysates and supernatants from cells transfected with pcDNA3.1-S and in the lysates from cells transfected with pcDNA3.1-GM-CSF-S.Conclusion:Two eukaryotic expression plasmids were constructed successfully and could express HBsAg.

Objective To improve the teaching quality of medical genetics. Methods A medi-cal genetics questionnaire survey was designed and conducted among 2012 grade undergraduates of Kunming Medical University. The results of the questionnaire were statistically analyzed by Kruskal-wallis test. Results The students felt that, there were differences in the importance and difficulty of teaching sections (Z=395.818,Z=117.786,P<0.001). The practical abilities were different after learn-ing (Z=139.364,P<0.001). Most of the students had strong demand for clinical practice and genetic counseling, there were differences in the degree of demand in students of different majors (Z=8.432, P=0.038). Conclusion Method of adjusting teaching content and method was proposed and adding clinical practice in the future was recommend to improve the teaching quality .

Aim To study anti-inflammatory and antibacterial effects of total flavones of oldenlandia diffusa Willd(FOD).Methods The models of dimethylbenzene-induced ear swollen in mice,turpentine-induced granuloma pouch in rats,paw edema caused by egg white in rats,increased vascular permeability by acetic acid in mice and the antibacterial activity in vitro were used to study FOD. Results FOD(15、30 and 60 mg?kg~-1 )show a marked suppressive effect on dimethylbenzene-induced ear swollen and increased vascular permeability in mice. In addition,FOD(12、24 and 48 mg?kg~-1 ) could reduce the paws edema and the granuloma pouch in rats.On the other hand,FOD had significally bacteriostatic effects against eight kinds of bacterial in vitro, in which effect on coccus is superior to that on bacillus.Conclusion FOD has anti-inflammatory and antibacterial effects.

OBJECTIVETo understand the relationship between the different replication status of HBV and mutations in core promoter in mother and child infected by mother-to-infant transmission.

METHODSCore promoter was amplified by PCR and cloned into pGEM-T vector with T-A cloning technique. The recombinant plasmid pGEM-PreS/S was confirmed by digestion with restriction enzyme Apa I and Sac I. Two clones were selected to sequence each patient.

RESULTSEvery pair of mother and child had same serotype and genotype and the homology of nucleotides encoding "a" determinant was 98 to 100%. The number of mutations in core promoter in patients with high replication status was more than that of low replication status. Mutations were distributed in BCP and Kunitz-type serine protease inhibitor region mainly. This difference was not associated with mother or child.

CONCLUSIONSThe different replication status of HBV is caused by mutations in core promoter in mother and child infected by mother-to-infant transmission and seems not to be associated with the development status.

Child ; DNA Replication ; DNA, Viral ; biosynthesis ; genetics ; Genotype ; Hepatitis B ; transmission ; virology ; Hepatitis B virus ; genetics ; Humans ; Infant ; Infectious Disease Transmission, Vertical ; Mothers ; Mutation ; Polymerase Chain Reaction ; Promoter Regions, Genetic

Objective To evaluate the effect of sildenafil on the expression of hexokinase-2 (HK-2) in lung tissues in a rat model of pulmonary arterial hypertension. Methods Thirty-two male Sprague-Dawley rats, aged 2 months, weighing 200-220 g, were allocated into 4 groups with 8 rats in each group using a random number table: control group ( group C), pulmonary arterial hypertension group ( group PAH), low-dose sildenafil group (group S1 ) and high-dose sildenafil group (group S2 ). The model of pul-monary arterial hypertension was established through combining left pneumonectomy with subcutaneous in-jection of 60 mg∕kg monocrotaline. Two percent sildenafil 30 and 50 mg∕kg were administered by intragastric gavage once a day for 3 consecutive weeks starting from 5 weeks after pneumonectomy in S1 and S2 groups, respectively. The chest was opened after the end of administration for measurement of mean pulmonary arte-rial pressure (mPAP) and right ventricular systolic pressure (RVSP). The hearts and lungs were excised for determination of the percentage of the thickness of tunica media of pulmonary arterioles, size of right ventricular cardiomyocytes, α-smooth muscle actin (α-SMA) expression (by immunohistochemistry) and expression of hypoxia-inducible factor-1α (HIF-1α) and HK-2 ( by Western blot). Results Compared with group C, the mPAP, RVSP and percentage of MT were significantly increased, the size of right ven-tricular cardiomyocytes was enlarged, and the expression of HIF-1α, HK-2 and α-SMA was up-regulated in PAH and S1 groups, and the RVSP and percentage of MT were significantly increased, the size of right ventricular cardiomyocytes was enlarged, and the expression of HIF-1α, HK-2 and α-SMA was up-regula-ted (P<0. 05), and no significant change was found in mPAP in group S2 (P>0. 05). Compared with group PAH, the percentage of MT was significantly decreased, the size of right ventricular cardiomyocytes was decreased, and the expression of HIF-1α, HK-2 and α-SMA was down-regulated in group S1 , and the mPAP, RVSP and percentage of MT were significantly decreased, the size of right ventricular cardiomyo-cytes was decreased, and the expression of HIF-1α, HK-2 and α-SMA was down-regulated in group S2 (P<0. 05). Conclusion The mechanism by which sildenafil inhibits proliferation of pulmonary vascular smooth muscle cells is related to inhibiting the expression of HK-2 in lung tissues in a rat model of pulmona-ry arterial hypertension.

Objective: To investigate the effect and mechanism of all-trans retinoic acid (ATRA) on leukemic cell line U937 cells with NPM1 mutation. Methods: Human acute myeloid leukemia cell line U937 was explored, NPM1 mutated (A type) plasmids were transfected into U937 to form stable clones A1 and A2, which were identified by Western blot and Co-immunoprecipitation. The cell proliferation was measured by methylthiazolyl tetrazolium bromide (MTT) ; cell cycle and cell apoptosis were explored by flow cytometric; cell colony formation was measured by microscope count, the molecular pathways related to cell proliferation were measured by Western blot. Results: ①The cell proliferations of mutant A1 and A2 were inhibited significantly by 52.6% and 35.8% (P<0.05) , respectively under ATRA exposure. ②The percentages of G₀/G₁ stage of mutant A1 and A2 increased by 20.1% and 35.8%, respectively under ATRA exposure. ③All the U937 leukemic cells were inhibited under ATRA exposure; the decreased percentages of vector, wild-type and mutant NPM1 cells were 32.7%, 57.9% and 90.9% respectively. ④p-ERK decreased obviously after ATRA exposure in NPM1 mutated leukemic cells. ⑤More mutant NPM1 cells inclined to apoptosis under the exposure of ATRA and cytotoxic drugs than cytotoxic drugs alone, meanwhile more cells apoptosis occurred when ATRA was administrated after cytotoxic drugs exposure. Conclusions ATRA could inhibit cell proliferation and colony formation, blocked the cell cycle in the G₀/G₁ stage accompanied by the significant reduction of p-ERK in U937 leukemic cells with NPM1 mutation. Besides, ATRA could synergize with drugs to suppress the leukemic cells survival more effectively when ATRA was administered after the cytotoxic drugs exposure in U937 leukemic cells with NPM1 mutation.

This study was aimed to reveal the species characteristics of Chinese patent medicines for antitussive ef-fect and provide references for developing new drugs. This research targeted Chinese patent medicines for antitussive effect which were included in the Pharmacopoeia of the People's Republic of China and the New National Chinese Patent Medicines as well as those characterized by keywords such as cough cure, cough alleviating, antitussive effect, cough, persistent cough. The analysis was made on the species characteristics, such as the number of Chinese patent medicines for antitussive effect, license number, ethnomedicine patent medicines, drugs for children use, protection of varieties of traditional Chinese medicine, the number of drugs, the generic names of drug, and drug forms. The results showed that 684 Chinese patent medicines for antitussive effect collected in this research had ac-counted for 8.60% of the total 7 260 of Chinese patent medicines. A total of 7 450 license numbers were approved, and 33% of the Chinese patent medicines shares one license number. One Chinese patent medicine owns 16.6 li-cense numbers on average. Ethnomedicine patent medicines had 3 Tibetan prescriptions such as the Shiwuwei Chenxiang pill and 4 Mongolian prescriptions, such as the Siwei Tumuxiang powder. Drugs for children accounted for 14%, including 9 forms. The type of the generic names of drug reached 16 and most of them originate from abbrevia-tions of the main drug in prescription. The number of drugs in prescription ranges from 8 to 16. Chinese patent medicines for antitussive effect involved 16 forms, of which the proportion of the use of solid preparation was higher than the liquid preparation. It was concluded that Chinese patent medicines for antitussive effect were characterized by such advantages such as a variety of species, various forms, the reasonable number of drugs, considerable medicine retail market share and drug for children use which can meet the clinical needs, and meanwhile some prob-lems, such as a lack of criteria for the generic names of drug, the homogenization of fierce competition, and inade-quacy of ethnomedicine patent medicines.

Objective:To systematically assess the risk factors for acute pancreatitis in pregnancy (APIP) in China in recent 10 years.Methods:Acute pancreatitis, pregnancy, risk factors and clinical features were used as search terms. Case-control studies on APIP in China were retrieved by computer from PubMed, Cochrane Library, Embase, CNKI, Wanfang, VIP and Chinese Biomedical Literature Database from December 2011 to December 2021. RevMan 5.4 software was utilized for Meta analysis.Results:A total of 1525 APIP patients (645 severe cases and 880 mild cases) were included in 26 literatures. Meta analysis showed that early pregnancy, late pregnancy, hyperlipidemia and biliary etiology were the risk factors for APIP. In the early pregnancy, 4.22% of patients were in the severe group and 10.43% were in the mild group; In the late pregnancy, 81.95% of the patients were in the severe group and 69.72% were in the mild group. The causes of hyperlipidemia accounted for 52.16% in the severe group and 22.69% in the mild group. The causes of biliary disease accounted for 27.96% in the severe group and 41.71% in the mild group. All the differences of the above indicators between groups were statistically significant (all P value <0.05). The age at onset, body mass index, serum C-reactive protein and triacylglycerol levels of severe patients were higher than those of mild patients, and were positively correlated with the severity of APIP ( MD values were 0.99, 65.97, 1.33, 6.44, 95% CI values were 0.03-1.96, 0.59-2.08, 23.43-108.50, 4.49-8.39, P values were 0.04, 0.0005, 0.002, <0.001). The levels of blood calcium and serum albumin were lower than those of mild patients, and were negatively correlated with the severity of APIP ( MD values were -0.37, -5.93, 95% CI values were -0.50--0.24, -9.11--2.75, P values were <0.001, 0.003). Conclusions:Acute pancreatitis can occur in all stages of pregnancy. Early pregnancy is mainly mild, and severe cases are more common in late pregnancy. Hypertriglyceridemic pancreatitis progresses rapidly, and tends to become more severe in China.

Objective To explore the expression and clinical significance of autophagy related proteins Beclin-1,microtubule-associated protein 1 light chain 3(LC3)in placenta tissues of pregnant women with fetal growth restric-tion(FGR).Methods A total of 40 pregnant women undergoing delivery due to FGR and 40 pregnant women un-dergoing normal delivery in the Second Affiliated Hospital of Zhengzhou University were enrolled as FGR group and healthy group between August 2022 and August 2023.The general clinical data in the two groups were collected.The levels of Beclin-1 and LC3 mRNA in placenta tissues were detected by PCR,and expressions of Beclin-1 and LC3 proteins were detected by immunohistochemistry.The differences in positive of Beclin-1 and LC3 proteins among patients with different clinical data were analyzed.Results The placental thickness,placental mass and neonatal weight in FGR group were lower than those in healthy group(P＜0.05).The mRNA levels of Beclin-1 and LC3 in FGR group were higher than those in healthy group(P＜0.05)and positive expression rates of Beclin-1 and LC3 proteins were significantly higher than those in healthy group(P＜0.05).There was significant difference in positive expression profile of Beclin-1 and LC3 proteins among patients with different placental thickness,placental mass and neonatal weight(P＜0.05).Conclusions The positive expressions of autophagy genes(Beclin-1 and LC3)are related to FGR,and their specific expression levels are closely related to fetal growth and development.

Objective:To evaluate the feasibility and clinical results of micro-dissected chimeric (II-MCh) perforator flap based on the descending branch of lateral circumflex femoral artery(d-LCFA) for repairing composite soft tissue defects of limbs.Methods:From April, 2011 to December, 2019, a total of 19 cases of composite soft tissue defects of limbs combined with deep cavity were repaired with II-MCh perforator flap or micro-dissected thin lobulated (III-MPCh) perforator flap based on d-LCFA. The flap was micro-dissected before cut off the perforator. The deep dead cavity was filled with muscle flap, and the superficial wound was repaired with micro-dissected thin perforator flap. The sizes of flaps were 6.0 cm×3.0 cm-33.0 cm×8.0 cm. The muscle flaps were cut with volumes of 5.0 cm×3.0 cm×1.0 cm-13.0 cm×10.0 cm×1.5 cm. Donor sites of the flap were closed directly. Postoperative reviews were assigned at 1, 3, 6, 9 and 12 months. The appearance, colour, texture and recurrence of infection of the flap were checked and recorded.Results:Sixteen flaps survived successfully without necrosis and with primary healing at the donor sites. Two flaps had venous occlusion within 24 hours after surgery and healed after surgical exploration and venous anastomosis. One flap had arterial crisis within 24 hours after surgery and healed after surgical exploration and arterial anastomosis. All patients entered follow-up for 8-36 months with an average of 16 months. All recipient sites achieved satisfactory appearance and function, and only linear scars left at the donor sites.Conclusion:The II-MCh perforator flap based on d-LCFA is a special modality of anterolateral thigh perforator flap, which achieves a satisfying appearance at the recipient site by micro-dissecting the flap within one procedure. This emerging technique can effectively fill the dead cavity, and is an effective method to repair complex soft tissue defects of limbs combined with deep cavity.