The main objective of human tissue engineering is to grow cells in vivo to promote proliferation and differentiation and replace, repair, maintain, or enhance the functions of impaired tissues and organs. Embryonic stem cells are the first selected seed cells in tissue engineering due to their unlimited proliferation capability and the potential to differentiate into different kinds of cells. In this review will give a sketch of recent advances in the preparation of induced pluripotent stem cells, which can differentiate into many other kinds of cells, by applying different transcript factors including Oct3/4, Sox2, c-myc, Klf4, Nanog and LIN28 to somatic ceils via retroviral transduction.

Objective Setting a rapid, simple and accurate gene diagnosis method for trisomy 21 syndrome. Methods 4 short tandem repearts (STRs) loci (D21S11, D21S1411, D21S1412, D21S1414) in and near the core region(21q22.1-21q22.2) on 21 chromosome of blooding samples from 8 normal people, 11 cases of trisomy 21 syndrome and 1 case of fetus umbilical core were analyzed and detected by polymerase chain reaction(PCR), polyacrylamide gel electrophories, and silver staining. Results 6/8? 7/8? 3/8?7/8 of normal people showed two bands with a DNA content ratio of 1∶1. 2/8?1/8?5/8?1/8 of normal people showed one band in 4 loci. 7/11?8/11?6/11?7/11 of all patients showed two bands with a DNA content ratio of 2∶1;3/11?2/11?2/11?3/11 showed three bands with a DNA content ratio of 1∶1∶1. 1/11?1/11?3/11?1/11 showed one band. The blooding sample of fetus showed two bands with a DNA content ratio of 2∶1 at 3 loci and one band at 1 locus. 11 patents and 1 fetus were identified as trisomy 21 syndrome by analysis combining with 4 STR loci. Conclusion The 4 selected STR loci have high polymorphism.4 polymorphotic STR is a valuable gene marker for diagnosis of trisomy 21. Trisomy 21 can be diagnosed rapidly and accurately within 24 hours by PCR.

OBJECTIVE:To observe the efficacy and safety of minocycline hydrochloride combined with tinidazole in the treat-ment of chronic periodontitis. METHODS:100 patients with chronic periodontitis were randomly divided into observation group 1, observation group 2,observation group 3 and control group. Patients in control group were given full-mouth dental scalers and sub-gingival clean scraping,then 0.9% Sodium chloride injection and 3% hydrogen peroxide were used to alternately wash periodontal pocket bottom. Based on the treatment,observation group 1 was injected Minocycline hydrochloride ointment to periodontal pocket bottom until slight overflow after saliva isolation and desiccation,once a week;observation group 2 was given Tinidazole buccal tablet 1 tablet in the buccal middle gingival surface,twice a day,observation group 3 was given Minocycline hydrochloride oint-ment+Tinidazole buccal tablets(the usage and dosage was the same with above-mentioned 2 groups). The treatment course was 4 weeks. The clinical efficacy,PLI,PAL,PPD,MD,GI before and after treatment and incidence of adverse reactions were observed. RESULTS:The total effective rate in observation group 3>observation group 1 and observation group 2>control group(P<0.05), however,there was no significant difference between observation group 1 and observation group 2(P>0.05). After treatment, PLI,PAL,PPD,MD and GI in all groups were significantly lower than before,and observation group 30.05). There were no obvious adverse reactions during treatment. CONCLUSIONS:Minocycline hy-drochloride combined with tinidazole has better efficacy than minocycline hydrochloride or tinidazole in the treatment of chronic periodontitis,with good safety.

Objective:To observe the clinical effect of regulating Conception Vessel and unblocking Governor Vessel by acupuncture combined with tuina for cerebral infarction. Methods:A total of 61 eligible cases with cerebral infarction were treated with regulating Conception Vessel and unblocking Governor Vessel by acupuncture combined with tuina, once a day, 12 d for a course. There was a 3-day interval between two courses. These cases were treated for 3 courses. Before treatment, the infarction severity was graded using neurological deficit score. The score changes and correlation between infarction severity and therapeutic efficacy were evaluated after 1, 2 and 3 courses of treatment. Results:Before treatment, the neurological deficit score was (18.65±3.28), versus (13.63±3.91), (9.53±3.22) and (5.57±3.97) respectively after 1, 2 and 3 courses of treatment. When the third course was completed, 4 cases obtained almost full recovery, 33 cases obtained marked effect, 18 cases obtained improvement and 6 cases had no effect. The total effective rate was 90.2%. Conclusion:Regulating Conception Vessel and unblocking Governor Vessel by acupuncture combined with tuina can substantially improve limb function in patients with hemiplegia due to cerebral infarction.

OBJECTIVE:To optimize the formulation of Cilnidipine sustained-release tablet,and study its drug-release mecha-nism. METHODS:Solvent method was adopted to prepare the cilnidipine solid dispersion,then Cilnidipine sustained-release tablet was prepared by using hypromellose K4M(HPMC K4M)as release material. Using comprehensive scores of cumulative release de-gree in 2,6,12 h as indexes,single factor method and Box-Behnken response surface method were used to screen the amounts of HPMC K4M and ethyl cellulose (EC),lactose-microcrystalline cellulose (MCC) ratio in formulation of Cilnidipine sustained-re-lease tablet,and verification test was conducted. The drug-release mechanism of Cilnidipine sustained-release tablet was investigat-ed by model fitting way. RESULTS:The optimal formulation was as follow as 25％ of cilnidipine solid dispersion,30％ of HPMC K4M,10％ of EC,lactose-MCC ratio of 1:1(m/m). The adhesive was 5％ PVPP ethanol solution and the lubricant was 0.5％magnesium stearate. The cumulative release degrees of prepared sustained-release tablet in 2,6,12 h were(21.4±3.3)％,(62.9± 2.8)％,(85.4±0.5)％(n=3),relative error of which to predicted value 25％,60％,90％were 14.4％,4.8％and 5.1％. Release curve showed the highest fitting degree with the first-order release model,conforming to non-Fick diffusion. CONCLUSIONS:Cil-nidipine sustained-release tablet with sustained-release effect is successfully prepared by optimized formulation.

Objective To explore the effect of abortion, effect of Yunnan Baiyao Combined with Wujia Shenghua capsule in patients with medical abortion vaginal bleeding time.MethodsSelect 98 cases from March 2014 to May 2016 by drug abortion in second hospital of shandong university, were randomly divided into combined group and control group with 49 cases in each group, two groups were given routine drug treatment, combined group was given Yunnan Baiyao combined with Wujia Shenghua capsule, comparing the two groups of patients with abortion, vaginal bleeding time.ResultsCombined group and control group differences in the complete abortion rate was not statistically significant, combinde group of pregnant sac discharge time is shorter than the control group (P<0.05);combined group with the duration of vaginal bleeding, vaginal bleeding and vaginal bleeding in 7~14 days>14 days the percentage of patients were lower than those in control group (P<0.05);combinde group of vaginal bleeding (16.33%), the amount of menstrual and menstrual amount (59.18%), > menstruation (24.49%) and the control group (12.24%, 38.78%, 46.94%) with significant difference (P<0.05);combined group of menses time, menstrual duration compared with the control group, the difference was not statistically meaning.ConclusionYunnan Baiyao combined with Wujia Shenghua capsule can accelerate the gestational sac discharge, shorten the time of vaginal bleeding, reduce the amount of vaginal bleeding.

Objective:To investigate the expression level and clinical value of serum miR-148a-3p and miR-551b-5p in patients with acute pancreatitis (AP).Methods:The clinical data of 152 patients with AP admitted to Danzhou people's Hospital from January 2017 to September 2020 were selected. According to the severity of their illness, they were divided into MAP group ( n=70), MSAP group ( n=40), and SAP group ( n=42). The SAP group was divided into survival group ( n=25) and death group ( n=17) according to the prognosis of the patients. Another 50 healthy people were selected as the control group. Fastening venous blood was collected on the day of onset, and the expression levels of serum miR-148a-3p and miR-551b-5p in each group were detected by real-time quantitative PCR. The receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated. The expression levels of serum miR-148a-3p and miR-551b-5p were analyzed to evaluate the prognosis of SAP. The correlation between serum miR-148a-3p and miR-551b-5p expression levels in SAP patients was analyzed by Pearson method. Results:The expression levels of serum miR-148a-3p (3.18±1.27 vs 0.96±0.28) and miR-551b-5p (1.94±0.85 vs 0.51±0.12) in AP group were significantly higher than those in control group ( P<0.001). The expression levels of serum miR-148a-3p (4.36±1.70 vs 2.84±1.10, 2.50±0.92) and miR-551b-5p (2.80±1.04 vs 1.68±0.53, 1.42±0.45) in SAP group were significantly higher than those in MSAP group and MAP group ( P<0.001). The expression levels of serum miR-148a-3p (5.30±1.95 vs 3.47±1.40) and miR-551b-5p (3.62±1.37 vs 2.08±0.91) in death group were significantly higher than those in survival group ( P<0.001). ROC curve analysis showed that the AUC of the combination of miR-148a-3p and miR-551b-5 in judging the prognosis of SAP was significantly higher than that of the single index of miR-148a-3p and miR-551b-5[0.943(95% CI0.886-0.997) vs 0.860(95% CI0.797-0.924), 0.822(95% CI0.795-0.880)], with a sensitivity of 98.3% and specificity of 84.6%. Correlation analysis showed that there were positive correlation between the expression level of serum miR-148a-3p and miR-551b-5p in SAP patients ( r=0.835, P<0.001). Conclusions:The expression levels of miR-148a-3p and miR-551b-5p in serum of AP patients were significantly increased, and the combined detection of miR-148a-3p and miR-551b-5p had a good value in judging the prognosis of SAP.

Objective To clone and express the outer surface protein C ( OspC) from a Chinese Borrelia afzelli FP1 strain and to evaluate the immune protectivity of the recombinant OspC protein ( rOspC) . Methods The gene encoding OspC protein of Borrelia afzelli FP1 strain was amplified by polymerase chain reaction (PCR) and then inserted into pET-30a plasmid to construct the recombinant expression plasmid pET-30a-OspC. The transformed E. coli BL21 strains carrying pET-30a-OspC plasmid were induced by IPTG to express OspC protein. The expressed proteins were purified by Ni-IDA resin chromatography and analyzed by SDS-PAGE and Western blot assay. Indirect immunofluorescence assay ( IFA) was performed to detect anti-rOspC protein antibodies in serum samples from rabbits immunized with rOspC protein. In vitro neutral-ization test was performed for evaluation the immune protectivity of rOspC protein. Results The recombi-nant expression plasmid pET-30a-OspC was successfully constructed and highly expressed in E. coli BL21. A strong antigen-antibody reaction between the rOspC protein and polyclonal antibody against Borrelia afzelli FP1 strain was detected by Western blot assay. The titers of IgG in serum samples from rabbits immunized with rOspC protein were significantly elevated. The in vitro neutralization test indicated that 106/ml of Borre-lia afzelli FP1 strains were neutralized by every anti-OspC protein serum sample from the experiment group. Conclusion The rOspC protein showed a strong immune protectivity against Borrelia afzelli, which could be used in the development of polyvalent subunit vaccine against lyme disease.

Rhizobium leguminosarum biov.trifolii H954 has been fermented for eight days on the GMS medium.Culture supernatant is extracted by ethanol precipitation,sample is purified by Sephadex G\|50,Sephadex G\|10 and DEAE\|Cellulose chromatoraphy.Gas chromatography(GC), 13 C nuclear magnetic resonance spectroscopy(NMR)and mass spectrometry(MS) analysis indicated that the sample is cyclic,neutral glucan,and composes of glucose linked solely by 1→2 glucosidic bonds.Cyclic glucan has degrees of polymerization ranging form 17 to 22,with a degree of ploymerization of 19 as the major glucan cycles.

Objective To explore the change of monocyte chemotactic factor-1 protein(MCP-1)and matrix metalloproteinase-9( MMP-9)of patients with coronary artery disease( CAD)following percutaneous coronary interventional( PCI). Methods Fifty patients underwent PCI procedures for CAD compromising a single coronary artery were selected as PCI group and 30 healthy individuals with normal findings by coronary angiography were selected as the control group. Plasma MCP-1 and MMP-9 were measured in all the subjects. Results The plasma MCP-1 level of patients with CAD after PCI was(19. 87 ± 5. 31)ng/ L,higher than that before operation((15. 71 ± 5. 23)ng/ L,t = 3. 95,P < 0. 01). Whereas in the control group,the MCP-1 level after coronary angiography was(13. 78 ± 5. 58)ng/ L,which was as same as that before operation (12. 42 ± 5. 39 ng/ L,P = 0. 34). Plasma MMP-9 level in the CAD patients after PCI procedures was(22. 69 ± 5. 97)mg/ L,higher than that before operation((19. 52 ± 5. 72)mg/ L,t = 2. 71,P < 0. 01). There was no significant difference in term of plasma MMP-9 level in control group befor and after operation((17. 53 ± 5. 51) mg/ L vs.(16. 69 ± 5. 42)mg/ L,P = 0. 55). Conclusion Plasma MCP-1 and MMP-9 increase in CAD patients following PCI procedures. But their roles in the vascular restenosis following the procedures need further investigation.